text:
Pyramids in georgia
9 Rated by Guests. mer-chefInches scuba diving dives towards the resort to make your food and assist the couple. or on H2. " The Pyramids of Giza. The Kenimer Site is an archaeological site near Sautee Nacoochee, Georgia in White County. Arches & Schwibbogens. When drawn as a "population pyramid," age distribution can hint at patterns of growth. Built between the 1st and 7th centuries A. Jan 21, 2019 The super blood wolf moon rises behind the Transamerica Pyramid in The totally eclipsed moon as seen from Athens, Georgia, Jan. In all the world, only the pyramids at Cholula and Teotihuacan in central Mexico surpass the Cahokia pyramid in size and total volume. On May 8, 2002, 300 local and federal law enforcement officers in rural Georgia raided the Egyptian-themed compound of the United Nuwaubian Nation of Moors. "It was written by Richard Thornton who writes articles on architecture and design for the Examiner's Web site, which pays writers' fees based on how many people view the ads on the Internet page. Is this the lost Mayan city of Yupaha? Historian creates 3D image of 1,000-year-old civilisation in GEORGIA mountains. With the red pyramid, Sneferu set the standard for all true pyramids to come. Special offer: Book during this week and get a free Camel ride at the Pyramids Area . I had visited there before 8 months with my family through a well known travel agency ask-Aladdin. This historically significant park is the oldest and largest Woodland Indian site in the southeastern United States, occupied by Indians from 350 to 750 A. D. Just drag the mouse to the direction you want to look. One of the main attractions of Egypt are Pyramids of Giza. com. 6 based on 59 Reviews "DJ China Doll" Also, among all indigenous peoples of the Americas, only the Itza Mayas and the ancestors of the Creek Indians in Georgia built five-side earthen pyramids as their principal mounds. The U. Georgia Tech Yellow Jackets. Order Here for Free Shipping! Courtesy: Baton Rouge Advocate The construction of millions of ancient monuments of earth, found all over South Intro to Art Chapters 6 - 10 study guide by abrama819 includes 100 questions covering vocabulary, terms and more. Ecological Pyramids Virtual The ancient Egyptians built Their pyramids as tombs for the pharaohs and their queens. The United Pyramids Lounge BAR - 2787 Main St, Atlanta, Georgia 30344 - Rated 4. Until now, Georgia was the only state to have a four-tiered pyramid. Visit reports, news, maps, directions and info on Nuwaubian Pyramids of Georgia in Eatonton, Georgia. The Steelcase Corporate Development Center near Grand Rapids, Michigan. Use the scroll wheel to zoom at the details. AGE DISTRIBUTION. World Archaeology 41 (3): 445-470. And it least you wouldn't find the right number. Once I acquired every thing jointly these people were being collateralized and equipped jointly creating a incredibly reliable headgear. Some states like Georgia prohibit pyramid schemes under a statutory framework that regulates business opportunities and multilevel marketing. From the mountains to the coast, there's something for everyone to explore in Georgia. Response to Intervention (RTI): The Georgia Student Achievement Pyramid of Interventions graphically illustrates the process of aligning appropriate assessment with purposeful instruction for all students. Home for 100% Handmade Orgone & Orgonite Pyramids made with Precious, Semi-Precious Stones, Gold, Silver, Copper, and other Natural Materials. It was 1969 and Habersham founder, Joyce Eddy, had just been given the chance to operate a small antique shop located above an old laundromat. I have always been fascinated by the ancient world and you don't get much more classically ancient than the Egyptian Pyramids. The Giza pyramids were constructed between 2589 and 2504 B. Pyramid Replacement Parts : About Us 1-888-968 According to a hypothesis proposed by author William "Pila" Chiles in his book 'The Secrets and Mysteries of Hawaii: A Call to the Soul', the pyramids may have been built on one of the Hawaiian Islands by an ancient culture and share a connection with other pyramids on Earth, through the configuration of a ancient planetary grid. . There are six step pyramids in the center, each with a temple on the top. They lynch me still in Mississippi. If it weren't for the pyramids, Visoko wouldn't have a whole lot going for it. Thompson The As demonstrated by its presence in the Egyptian pyramids, gypsum has served as a vital piece of many famous construction projects throughout history. So, with that said, we can carry on. org It's a new day on the internet so there's a new conspiracy theory: a mysterious new pyramid has been discovered in Antarctica thanks to Google Earth. . The Nation of Nuwaubia is gone. Crafter Meroe, an archaeological site in Sudan, is the location of the numerous pyramids of the Kushite Kingdom built in the 3 century BCE. The Pyramids virtual tours can be started by clicking the preview window or by downloading the stand-alone versions. He included aboveground burial chambers, a mortuary temple, and a causeway leading down to a valley temple. View Lab Report - Ecological Pyramids Virtual Lab Answer Key BIO 124 Georgia Military College Online from BIO 124 at Georgia Military College. Six hours southeast of Atlanta off the Georgia coast on Sapelo Island, archaeologists have unearthed the remains of an ancient walled city which predates the construction of Egypt's pyramids. According to Richard Thornton at Examiner. Trust me, the site is massive! If you're short on time or just want to see the main sights, this is what to look for: There are at least three ancient underwater pyramids off the Atlantic Coast of Florida which few people know exist. A University of Georgia Brasstown Bald Etowah Mounds Mayan in Georgia Etowah mounds in Cartersville, Georgia. Brad from Georgia's Avatar · Brad from Georgia Brad from Georgia is offline. Visitor Tips, news, stories, field reports. Georgia. Testimonials. Welcome to the state's official tourism website! We're here to help you plan your Peach State adventure and keep Georgia on Your Mind. 29 Reviews (404) 458-3973 Website. The $5 note issued by Georgia in 1777 features a coiled rattlesnake and the Latin motto Jun 28, 2017 Located in southern Turkey, this ancient place of ritual worship predates Stonehenge by 6,000 years and the first Egyptian pyramids by 5,000 . Media in category "Population pyramids of counties of Georgia (U. When we say pyramids, the Giza Pyramids are the ones that always come into mind. " Georgia Department of Education Georgia Standards of Excellence Framework GSE Grade 6 Mathematics • Unit 5 Mathematics Grade 6 Unit 5: Area and Volume July 2019 Page 7 of 108 • What kinds of problems can be solved using surface areas of rectangular and triangular prisms and pyramids? The Ministry of Antiquities announced important discoveries at a recent press conference, held on the eastern side of the Great Pyramid of Khufu on the Giza Plateau. The project hopes to discover the secrets of the Pyramid Get directions, reviews and information for Pyramids Lounge BAR in Atlanta, GA. I've been a victim: The Belgians cut off my hands in the Congo. The government seized the Egyptian-themed Archaeologists have discovered the ruins of an ancient Mayan city in the mountains of North Georgia believed to be at least 1,100 years old. Georgia's Indians carved marble statues and housed them in temples built atop multi-story earthen pyramids. ) You can watch it on Youtube. , building magnificent temples in Guatemala, Honduras, Belize and southern Mexico. I've been a singer: All the way from Africa to Georgia I carried my sorrow songs. Pyramid Candles. Known as the Sapelo Shell Ring Complex, this ancient The pyramids at Giza are part of the largest remaining necropolis, which included funerary temples as well as the Sphinx. Need more info? Feel free to give us a call at 1-800-VISITGA (847-4842) or drop us a line at travel@exploregeorgia. Journey back in time as you explore giant stone temples and mural-adorned palaces in the ancient city of Teotihuacan, a UNESCO World Heritage Site located approximately 30 miles northeast of Mexico City. Pyramids Lounge BAR - 2787 Main St, Atlanta, Georgia 30344 - Rated 4. No matter who you are a person who has never interested in pyramids or you have PhD in History, it's not matter you do not find an answer on this question. Pyramids Lounge BAR 2787 Main St Atlanta GA 30344. Ok, so the Georgia Guidestones aren't that ancient, but still, they are fascinating. Albany, Georgia 31708. Predating the arrival of humans in North America and the pyramids of ancient Egypt by more than 50,000 years, the trees discovered in the Underwater Forest Jun 22, 2019 Your preparation for visiting Pyramids starts here. Students will Nov 23, 2015 It might be hard to bee-lieve but the 3000-year-old sweet treat is still edible. Jul 6, 2018 Dwight "Malachi" York with his followers in Putnam County, Georgia, stucco pyramids and a Sphinx on a former 476-acre game preserve. The Pyramids - 3D Virtual Tour. Teotihuacán is a huge UNESCO site with a 2,000 year-old history. Size and geometric perfection seem to be less important than religious and mathematical function. At the Commission, we bring cases against pyramid schemes under the FTC Act, which broadly prohibits "unfair or deceptive acts or practices in or affecting commerce. S. Ziggurat (below) Teotihuacan Author acarioli Posted on 4 September 2017 14 August 2017 Categories ggplot2, R Tags Caucasus, Georgia, population pyramids Post navigation Previous Previous post: Long to wide format with tidyr (and save it in n files) EATONTON, Ga. Tales about the Bermuda Triangle (also known as the Devil's Triangle), a region in the western part of the North Atlantic Ocean where The World's Oldest Papyrus and What It Can Tell Us About the Great Pyramids Ancient Egyptians leveraged a massive shipping, mining and farming economy to propel their civilization forward The Stonehenge-like granite slabs, known as the Georgia Guidestones, have sparked controversy around the world and their meaning is still up for debate. state)" The following 159 files are in this category, out of 159 total. Georgia is known for welcoming a wide variety of food into its cultural palate. Cairo Pyramids and Saqqara day trip. Archaeological excavation, carried out intermittently at the site for more than a hundred years, has provided much information about the prehistoric life of the town. Know more about the pyramids with fact file or worksheet collection. Explore the inspiring Pyramids Tour in Egypt! Get amazed by Pyramid tour in Cairo discovering the ancient history of the pharaohs. HotelsCombined. No pyramids are more celebrated than the Great Pyramids of Giza, located on a plateau on the west bank of the Nile River, on the outskirts of modern-day Cairo. A free inside look at company reviews and salaries posted anonymously by employees. It has reduced in height a little since it was built and is currently 138. Now, Georgia operates within a three-tiered, multi-level prevention system. Jan 30, 2012 Six hours southeast of Atlanta off the Georgia coast on Sapelo Island, archaeologists have unearthed the remains of an ancient walled city Jan 8, 2012 Around 200 AD Indians in Florida and Georgia began the construction of enormous earthen pyramids. Ocmulgee National Park - Macon, Georgia . The monumental tombs are relics of Egypt's Old Kingdom era and were constructed some 4,500 years ago. Once these pieces manifest in full brilliance, they begin to "talk" to one another, too, through the beautiful layering of colors, textures and themes. The Mississippian Production of Space Through Earthen Pyramids and Public Buildings on the Georgia Coast, USA. Jan 13, 2013 I think this is a new History Channel series and thought this video might interest some of you guys. The pyramids of Egypt are part of everyone's travel imaginary and recognized worldwide as the symbol of Egypt and its ancient civilization. But, these amazing manmade infrastructures aren't just found in Egypt. ChartGraphics -- Age-Gender-Race- Origin Analysis & Population Pyramids . Pyramids of Giza that you want to visit are the oldest and largest of the three pyramids in the Giza. Egypt's pharaohs expected Book Pyramids in Florida, Fort Myers on TripAdvisor: See 81 traveler reviews, 157 candid photos, and great deals for Pyramids in Florida, ranked #1 of 16 specialty lodging in Fort Myers and rated 4. But the locals seem to have rallied around the idea – there are souvenir shops selling keychains and gemstones, paperweights shaped like pyramids and all kinds of new age healing trinkets. the ancient Egyptian pyramids of king Khufu, Khafre and Menkaure, built in that order, are a testament to ancient planning and engineering. It was Pyramids in Florida. This was just the opportunity a woman of Joyce's vision and energy would turn into the perfect blend of utility, artistry and soul. It's the oldest of the Seven Wonders of the Ancient World and the only one to still remain intact. The Daily Debunker brings you the top stories on Snopes. The stunning Great Pyramids, enigmatic Sphinx, and treasure-packed Egyptian Museum await us in Cairo, and in Luxor, we walk among the stone columns of Aug 20, 2009 The Mississippian production of space through earthen pyramids and public buildings on the Georgia coast, USA. 6 based on 59 Reviews "DJ China Doll" Population Pyramids: Georgia - 2017. A top heavy pyramid, like the one for Grant County, North Dakota, suggests negative population growth that might be due to any number of factors, including high death rates, low birth rates, and increased emigration from the area. I don't have a concrete first memory of my desire to visit Egypt. Pyramid building belonging to The Digital Group (TDG), at Hinjwadi, Pune, India. Each piece is a work of art, handmade of sand cast aluminum with our signature blend of enamel infused with crushed Mother-of-Pearl. In Georgia, Response to Intervention is based in the general education classroom where teachers routinely implement a strong and rigorous Far from the dry and desolate deserts of Egypt, several pyramids can be found throughout the United States. Jan 2, 2016 Author information: (1)Institute of Plant Breeding, Genetics and Genomics, University of Georgia, 111 Riverbend Rd. Except it's just a mountain. Media in category "Population pyramids of Georgia" The following 10 files are in this category, out of 10 total. This site is located just miles from the previously mentioned source of "Maya Blue"pigment. Referred to by many as an "American Stonehenge," the Georgia Guidestones is a granite monument in Elbert County, Georgia with carved inscriptions in four ancient languages: cuneiform Babylonian, Classical Greek, Sanskrit and Egyptian hieroglyphs. At the same time corn also arrived in Jan 5, 2012 Could there be Mayan ruins in North Georgia? A researcher there wrote online that ancient ruins in the area were remarkably similar to Mayan Nov 2, 2017 A void as tall as the Statue of Liberty has been detected in Khufu's pyramid, otherwise known as the Great Pyramid, using a technique of Sep 1, 2004 Critics of the Bible say the pyramids were built long before Noah's Flood, so the Flood must have only been a local affair, not global. The town features various locations linked to the Bosnian Pyramids theory. At the same time corn also arrived in Florida. The Teotihuacán built the Pyramids of the Sun and of the Moon between A. How many pyramids do you think there are in egypt? There is 138 pyramids in egypt They have to use over 5000. goverment -- which seized the place in a criminal forfeiture action -- sold it in Spring 2005 to a local developer. And even the America Unearthed: American Mayan Secrets (review/commentary) American Mayan Secrets is the first episode of the series America Unearthed , originally aired right on 12/21/2012. Mayan Pottery, Pyramids Unearthed in Georgia New research reveals that symbols which appear on ancient Georgia pottery are identical to Mayan glyphs, the symbols used to write the Mayan language. Corn is a native crop of Mexico and archaeologist William Sears, who first discovered this evidence Central American pyramids are closely related to the ziggurats of Sumer, Babylonia and Assyria: they are step pyramids with a central staircase, and a shrine at the top. We have woven into every detail of the luxury brand's service and design an acknowledgement of Chinese culture and heritage. November 2019: Georgia Aug 19, 2012 Planetary alignment over the Giza pyramids Science. Centuries of research have given us parts of the answer. 2015: In the Shadow of the Pyramids (self published, edition of 500 / ISBN ABJ/ BA journalism & political science, University of Georgia - Athens, Georgia, USA Fairfax, VA – Congratulations to Jeff Hwang, CEO & Co-Founder, and Sherry Hwang, President & Co-Founder, of Pyramid Systems, Inc. we offers fabulous varieties of classical tours in Egypt, covering the most magnificent Our Logoregions including Cairo, Luxor, Aswan, Abu Simbel, Alexandria and Diving in Red Sea, Nile Cruises in luxor, dahabiya nile cruise, Shore Excursion From Safag Port 2 Explore the Egyptian Pyramids 3 Ski in Georgia (the Country, not the State) 4 Sleep in a Ger in Mongolia 5 Go Mushing in Alaska 6 Hike Across England 7 Cruise for Boobies in the Galapagos Scroll for more The Kolomoki Pyramids, Georgia United States - 350 AD - 600 AD Temple Pyramid, Witherville, Mississippi United States -1000 AD Pyramid (Mound) A, Poverty Point, West Carroll, Louisiana United States - 1650 BC - 700 BC - This is the largest of the five pyramids in the area. The cult's founder, Dwight "Malachi Pyramids bar and hookah lounge has been in business for 3 years. 1 and 250. No other structure in the United States approached the size of the Cahokia pyramid until the building of airplane hangars, the Pentagon, and skyscrapers in the twentieth century. Fairfax, VA – Congratulations to Jeff Hwang, CEO & Co-Founder, and Sherry Hwang, President & Co-Founder, of Pyramid Systems, Inc. Victor D. 45mins north east of downtown Atlanta, 90 mins from Braatown Bald Mountain. Georgia's oldest great temple mound, standing 57-feet high, dominates two smaller burial mounds and several ceremonial mounds. , who were recognized with Distinguished Alumni Awards by the Alumni Association of Georgia State University at its 52nd Annual Distinguished Alumni Awards Gala on Friday, October 2, 2015. Georgia Ancient Ruins: See reviews and photos of 10 ancient ruins in Georgia, Europe on TripAdvisor. Built as massive It all started in the small North Georgia town of Clarkesville. The Pyramids of the City Stars Complex in Cairo, Egypt. Experience Street View, 3D Mapping, turn-by-turn directions, indoor maps and more across your devices. 8. Under my hand the pyramids arose. Hawaii, 2000 Show All; All Pyramids; All Stunts; DOD Questions; Section 1 - Cheerleader Apparel/ Accessories; Section 3 - Inversions; Section 4 - Non-release Stunts; Section Richard hires Ling, and Billy, Ally and Georgia want to band together so they don' t lose the firm to Ling, but Billy and Ally seem to be forming their own coalition Food pyramids and food plates were developed as a tool to help you eat better and suggest healthy foods in healthy portions to form the foundation The majority of food pyramids and plates suggest eating peanuts. Response to Intervention (RTI) Georgia's Tiered System of Supports for Students (Georgia's MTSS) is a framework that integrates assessment and intervention within a school-wide, multi-level prevention system to maximize student achievement and reduce behavioral problems. We offer a large variety of elite hookahs that are prepared with care and offering the most flavorful tobaccao. There are about 100 altogether, some only symbolic and small. we offers fabulous varieties of classical tours in Egypt, covering the most magnificent Our Logoregions including Cairo, Luxor, Aswan, Abu Simbel, Alexandria and Diving in Red Sea, Nile Cruises in luxor, dahabiya nile cruise, Shore Excursion From Safag Port Love Egypt Tours is trustworthy travel agency in Egypt. City roadtrip recommendations: Atlanta | Columbus The mystery of how, exactly, the pyramids were built may have come a step closer to being unravelled after a team of archaeologists made a chance discovery in an ancient Egyptian quarry. Multiple sources of information about the pyramids are available, but for simplicity most of the following information is from Bill Hanson's book "The Atlantis Triangle. "Lost Worlds: Georgia" hopes to dispel many of the outdated stereotypes forged by Mayan Pyramids in Georgia? One of the largest collections of Swift Creek pottery was unearthed at the Kolomoki Mounds site in southwest Georgia. Nature reigns superior with this Arctic wild. German Pyramid Carousels. Mailing List - See more. Around 200 AD Indians in Florida and Georgia began the construction of enormous earthen pyramids. HUALUXE® Hotels and Resorts is the first upscale international hotel brand designed specifically for Chinese guests. net Population Pyramids of the World from 1950 to 2100. Quizlet flashcards, activities and games help you improve your grades. Drug Rehab Centers In Georgia. Join LinkedIn today for free. Pyramids in America . Join Date: The reverse of the Great Seal features an unfinished pyramid, which . 8 meters high (455'). com, the Nuwaubian Pyramids of Georgia. Find 48 listings related to Pyramids in Atlanta on YP. Georgia, 2000 · 2010. Georgia cuisine ranges so Its occupation continued, with brief periods of abandonment, into the seventeenth century. Between the pyramids and the museum, there's a ton to see and you can easily spend a whole day there. Reviews I've been watching America Unearthed (H2 channel) since the early episodes, including the very first one where they claimed to have found Mayan pyramids and artifacts in Georgia. But there are 17 great pyramids, and the size and composition of these stagger the minds of those who visit them. The Giza Pyramids, built to endure an eternity, have done just that. These structures were built by a chain of successive pharaohs in a line from grandfather to great-grandson. Now owned by the state of Georgia, the site and an interpretive museum is open to visitors. PopulationPyramid. The pottery known as Swift Creek was a highly decorated form of pottery produced around 2,000 years ago beginning around 0 A. See reviews, photos, directions, phone numbers and more for Pyramids locations in Atlanta, GA. Get Animated Pyramids Click icon to create Animated Pyramid Charts that show the age / race / sex distribution of counts or percentages, including associated data tables. There are six pyramids in the complex but we mostly think of this one, commonly known as the Great Pyramid since it is the largest. I am a Negro: Black as the night is black, Black like the depths of The last remaining wonder of the ancient world; for nearly 4000 years, the extraordinary shape, impeccable geometry and sheer bulk of the Giza Pyramids have invited the obvious questions: 'How were we built, and why?'. , Athens, GA, 30602, USA. The site contains two earthwork mounds located on top of a The Nuwaubian pyramids were constructed in the early 1990s by the Nuwaubian quasi-religious cult at the cult's Tama-Re compound in Eatonton, Georgia. The place In . (I didn't think to review it at the time. Discover the world with Google Maps. It followed release of the preliminary results of the Scan Pyramids project. It's been bulldozed down to the bare earth. Not all of the structures have been excavated yet! Currently, the cleared area of the ancient city covers only 16 square kilometers, but there is a lot to see there (no wonder UNESCO included Tikal in its World Heritage List). I made ragtime. Pyramids Construction Contractors Inc, Savannah, GA holds a Residential/General Contractor license according to the Georgia license board. Pyramids in Florida and Georgia Constructed by Mayans? Artist's interpretation of how the great earthen pyramid at Letchworth Mounds would have appeared around 400 AD. This was the model followed by his son, Khufu, wh built the first and largest pyramid at Giza. Georgia Attractions and Oddities. BLUE Restaurant and Hookah Lounge Learn about working at Pyramid Consulting, Inc. Pyramids in Florida ™ 7020 Constitution Loop, Fort The Great Pyramids of Giza . Most people associate the pyramids of Egypt with the Giza pyramid complex near Cairo, but truth is that Egypt has over 100 pyramids! You can find pyramids on all continents on Earth and in many places under water. Rock terraces and mounds by North Georgia mountain date to 900AD Archeologists Georgia are claiming that they have discovered some ancient Mayan ruins. We can seriously say that Monk Mound is a pyramid, which doesn't looks like the pyramids in Egypt or in Mexico; one can see some similarities with Chinese pyramids because in both cases the pyramids were constructed from the soil. No testimonials have been added yet. Almost everyone has been taught that the Mayan people thrived in Central America from about 250 to 900 A. " Love Egypt Tours is trustworthy travel agency in Egypt. Discover more Bars / Taverns companies in It's a long magical hike to the Tepozteco Pyramid, one of the many Aztec Pyramids in Mexico, but this one is situated on top of the mountain overlooking the Mar 19, 2018 FROM "proof" of UFO crashes to Antarctic pyramids, conspiracy cylindrical object discovered on the remote island of South Georgia was the Cradle: A dismount from a partner stunt, pyramid or toss in which the catch is Examples of hanging pyramids are: a person being suspended between two First, just take a look at some of the ancient pyramids around the world and when they were The Kolomoki Pyramids, Georgia United States - 350 AD - 600 AD. Seeing the Egyptian Pyramids was one of those dreams I had harbored as long as I can remember. Their BuildZoom score of 90 indicates that they are licensed or registered but we do not have additional information about them. All the weird attractions, hidden sights, and unusual places in Georgia. The Monk Mound in Illinois, USA is one of the biggest pyramid constructions in the world. When a black supremacist cult leader moved to rural Georgia, folks took a live-and-let-live approach — until Dwight York and his United Nuwaubian Nation of Moors launched an all-out campaign against "white devils" and "house niggers. Mar 28, 2013 The World's 15 Most Amazing Pyramids (PHOTOS) high step pyramid was built in stages, made from ancient Egyptian tombs . Some are an overt homage to the Northern African country and its identifiable architecture. The December 21, 2011 headline of the article put online by the Examiner declared: "Ruins in Georgia mountains show evidence of Maya connection. Egypt is really one of the best places to visit on the entire planet and pyramids are its ornaments. What to see at Teotihuacan Pyramids. Information Technology and ServicesAlpharetta, Georgia46,889 followers. Temple Mounds & Pyramids In The Americas . It was 1969 and Habersham founder, Joyce Eddy, had just been given the chance to operate a small Sep 4, 2017 You can download session 9 files for constructing the population pyramids of Georgia here: RLadies Tbilisi And with scales="free_x" Population Pyramids; CHartGraphics. Get an unparalleled tour to Cairo Pyramids and sakkara After meeting your Egypt Travel Vactions guide at your hotel in Cairo will transfer you by A/C vehicle to start your day trip your first stop will be at Giza Pyramids first of all you will face the pyramids of khufu If you ask yourself how many pyramids are in the world you will be lost in thoughts. Could there be Mayan ruins in North Georgia? A researcher there wrote online that ancient ruins in the area were remarkably similar to Mayan structures in Central America. From Pyramids to Clouds [4600 - 4999] Cambridge Dr Atlanta GA 30338. C. Ancient Egyptians By Georgia & Sophie w. Like many Mesoamerican pyramids, each was constructed around a core of rubble held in place by retaining walls. Contrary to what mainstream science claims, they are not created by using slaves and ropes, but by alien visitors and by ancient peoples who had learned the technique from extraterrestrial and interdimensional beings, like the Atlanteans. Six hours southeast of Atlanta off the Georgia coast on Sapelo Island, archaeologists have unearthed the remains of an ancient walled city which predates the construction of Egypt's pyramids. This is the only underground earthlodge that has been rebuilt to date. The pyramids of Egypt are amazing structures and fascinate people today. — Pyramids, obelisks and a lonely sphinx stand deserted on the Egyptian-themed compound where as many as 500 members of a quasi-religious sect lived only five years ago. Honestly, after watching a season and a half of this show, it is not the greatest pseudo-documentary. Offering a comfortable, relaxing, and uptempo atmosphere with a live dj to give our clients the best experience posssible. Find out More! Get information, directions, products, services, phone numbers, and reviews on Pyramids Lounge in East Point, GA. "Blue is a Mediterranean style, Hookah, Bar and Lounge restaurant located in the heart of Buckhead in Atlanta, Georgia. Click icon to create Maps by County, Public Health Districts and Perinatal Region. The group has taken numerous names, including Ansaru Allah Community, Holy Tabernacle Ministries, United Nuwaubian Nation of Moors (after the move to Georgia), Yamassee Native American Moors of the Creek Nation (also used in Georgia when York claimed indigenous ancestry via Egyptian migration and intermarriage with the ancient Olmec) and Small pyramids similar to those of the Louvre can be found outside the lobby of the Citicorp Building in Long Island City, Queens NY. This is a warning: Whoever reads this list of the most iconic foods in Georgia will be left with a severe craving for one (or more) of the items mentioned. 5 of 5 at TripAdvisor. , who were recognized It all started in the small North Georgia town of Clarkesville. Georgia Org. " Get directions, reviews and information for From Pyramids to Clouds in Atlanta, GA. Pyramids and Arches. I made mortar for the Woolworth Building. The Nuwaubian pyramids were constructed in the early 1990s by the Nuwaubian quasi-religious cult at the cult's Tama-Re compound in Eatonton, Georgia. , the holy city of Teotihuacan ("the place where the The Tama-Re village in Putnam County, Georgia was an Egyptian-themed set of buildings and The two pyramids are distinct in appearance and in usage. I shorten the video some (not any important 2346 American Income Life reviews. pyramids in georgia


Localization is pretty much essential for anyone whose native language is not among the ones built in the original device configuration. Even if you know English or German or Spanish well enough, if you are a native Serbian speaker, you would likely be receiving emails in Serbian, reading Serbian websites and having contacts in Serbian. Therefore, you would likely need to have Serbian localization for Palm, not just an application-by-application solution. Without attempting to completely turn the device to Serbian, InterType addresses the very need to be able to read and write other languages in every application.


The purpose of present work was to formulate and evaluate atenololtartrate floating microspheres. Floating Microspheres concept was applied to increase the gastric residence of the dosage form. The Floating Microspheres were prepared by emulsification solvent diffusion technique. The best batch exhibited excellent floating time as well as release at desired time. The particle size was controlled by changing polymer concentration, rpm and temperature. Polymers used for the preparation were Ethyl cellulose and HPMC. This approach suggested the use of floating microsphere to avoid the side effects. Keywords: Microspheres, Atenololtartrate, Ethyl cellulose, HPMC.
IAJPS 2018, 05 (01), 135-140 L. Satyanarayana and N.
approach suggested the use of floating microsphere to avoid the side effects.
Keywords: Microspheres, Atenololtartrate, Ethyl cellulose, HPMC.
Edulabad, Ghatkesar, Medchal, Telangana, INDIA.
Atenolol Floating Microspheres, Indo Am. J. P. Sci, 2018; 05(01).
Floating systems are the low-density systems that [2,3].
retention time and reduces fluctuation in the release formulation .
drug dispersed or dissolved throughout particle period, bioavailability enhancement.
molecules that exhibit absorption window, less speed and temperature as shown in Table 1.
Evaluation of atenololtartarate floating filtered then assayed by UV-vis spectroscopy.
1. Particle size determination drug content to theoretical drug content.
desiccator until constant weight was achieved.
crushed and then dissolved in distilled water with run in triplicate.
130.5 to 240.6 μm with increases the EC conc 1 to the temperature 450 ,600.
maximum up to 94% as shown in Table 12. available is more for drug release.
for more than 10 hrs due to insolubility of EC optimum particle size.
longer the floating time shown in Table 3. Gastro retentive Drug Delivery Systems. Arch.
Apll. Sci. Res, 2010; 5(2): 257-270.
drug release after 12 hours from microsphere JC, Encyclopedia of Pharmaceutical Technology.
concentration. It can also be seen that for 280-285.
the RPM increased the particle size increased Pharmainfo.net. 2006; 4(3): 146-152.
which leads to decrease in drug release 4.Cupp M . Alternatives for Metoprolol Succinate.
6.Biffi. M Boriani.G , Sabbatani.P , Bronzetti. G , Journal of Drug Delivery. 2010; 2(1):64-68.
7.Kannan.K, Karar.K.P, Manavalan.R. 12.Fischer S, Foreg C, Merkle P.H, Gander B.
pharmaceutical Sciences. 2009; 71(6): 715–718. blocker. EJPMR. 2017;4(04); 277-282.
Microcapsules: Effect of Solvents. Indian Journal of mouth dissolving tablets of carvedilol.
of pharmaceutical Sciences. 2003; 65(1):62-66. 2017;6(4); 518-524.


text:
Developmentally Regulated Oscillations in the Expression of UV Repair Genes in a Soilborne Plant Pathogen Dictate UV Repair Efficiency and Survival
Shira Milo-Cochavi, Sheera Adar, Shay Covo
Judith Berman, Editor
Shira Milo-Cochavi
Department of Plant Pathology and Microbiology, Robert H. Smith Faculty of Agriculture Food and Environment, Hebrew University of Jerusalem, Rehovot, Israel
Sheera Adar
Department of Microbiology and Molecular Genetics, The Institute for Medical Research Israel-Canada, Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel
Shay Covo
Judith Berman
The ability to withstand UV damage shapes the ecology of microbes. While mechanisms of UV tolerance were extensively investigated in microorganisms regularly exposed to the sun, far less is known about UV repair of soilborne microorganisms. Fusarium oxysporum is a soilborne fungal plant pathogen that is resistant to UV light. We hypothesized that its UV repair capacity is induced to deal with irregular sun exposure. Unlike the SOS paradigm, our analysis revealed only sporadic increases and even decreases in UV repair gene expression following UVC irradiation or exposure to visible light. Strikingly, a major factor determining the expression of UV repair genes was the developmental status of the fungus. At the early stages of germination, the expression of photolyase increased while the expression of UV endonuclease decreased, and then the trend was reversed. These gene expression oscillations were dependent on cell cycle progression. Consequently, the contribution of photoreactivation to UV repair and survival was stronger at the beginning of germination than later when a filament was established. F. oxysporum germinates following cues from the host. Early on in germination, it is most vulnerable to UV; when the filament is established, the pathogen is protected from the sun because it is already within the host tissue.
IMPORTANCE Fusarium oxysporum infects plants through the roots and therefore is not exposed to the sun regularly. However, the ability to survive sun exposure expands the distribution of the population. UV from the sun is toxic and mutagenic, and to survive sun exposure, fungi encode several DNA repair mechanisms. We found that Fusarium oxysporum has a gene expression program that activates photolyase at the first hours of germination when the pathogen is not established in the plant tissue. Later on, the expression of photolyase decreases, and the expression of a light-independent UV repair mechanism increases. We suggest a novel point of view to a very fundamental question of how soilborne microorganisms defend themselves against sudden UV exposure.
UV light is probably the most important environmental genotoxic agent. Microorganisms that are exposed to strong UV radiation, such as marine Flavobacteriia or Acinetobacter strains that live at high altitude, show diverse and efficient UV repair or damage avoidance mechanisms (1, 2). Far less is known about the DNA damage response of microorganisms that are not regularly exposed to UV. Escherichia coli is the model organism to study the repair of UV damage in organisms that are not exposed constantly to light. It has a sophisticated mechanism to induce the expression of UV repair genes known as the SOS system that may contribute to its ability to survive in diverse ecological niches (3–5). Fusarium oxysporum, the focus of this work, is an economically important fungal plant pathogen; one of the subspecies of this fungus endangers banana growth worldwide (6–8). F. oxysporum is a soilborne pathogen that penetrates plants through their roots. Therefore, no sun exposure is expected during most or even all of its life cycle. Nevertheless, F. oxysporum can survive high doses of UV (9). F. oxysporum has three types of spores (10–12). Microconidia are probably the most common type of F. oxysporum spore. Microconidia are root propagules that are found in the soil, although their occasional exposure to the sun is possible. The two other types are chlamydospores and macroconidia. Chlamydospores are heavily melanized resting units that can survive in the ground for years (10). These spores are expected to be UV resistant. Macroconidia are multinucleate spores that are usually formed on plant stems or leaves (10). Here, we focus our analysis on microconidia; the relevance of our results to other types of spores is discussed. Conidia germinate in response to plant signals, such as amino acids and plant peroxidases, but also in response to humidity and other nutrients (13, 14). Taking the disease cycle of F. oxysporum into account, the UV response of the fungus can be divided in two; the UV damage induction probably occurs during conidial germination, but development of the germlings into hyphae probably occurs already within the plant tissue and therefore away from UV and visible light (15).
The most common UV lesions are cyclobutane pyrimidine dimers (CPDs) and 6,4 UV photoproducts, both of which significantly distort the DNA helix. This severe disruption of the DNA structure interrupts DNA replication and transcription, thereby threatening the survival of microbes. UV is also mutagenic; it may cause deleterious mutations in the surviving population. Microorganisms have developed several mechanisms to deal with UV exposure (16–18). Ascomycete filamentous fungi are known for their remarkable ability to sustain UV exposure due to the function of three DNA repair mechanisms, nucleotide excision repair (NER), UV damage endonuclease (UVDE), and photolyase (19–21). NER does not recognize UV specifically; rather, it recognizes distortion of the DNA helix by scanning the genome (global NER) using the proteins Ddb1 and Xpc (22–24). Alternatively, the lesions are recognized by stalled RNA polymerase II with the assistance of proteins Csa and Csb, a process known as transcription-coupled NER (22, 23, 25, 26). After damage detection, Xpa, Xpb, and Xpd facilitate incisions on both sides of the lesion by two nucleases, Xpg and Xpf. The oligonucleotide containing the lesion is removed, and the gap is filled by DNA polymerases (22, 25). UVDE directly recognizes the two major UV lesions and makes a nick in the 5ʹ direction from them that is then further processed (27, 28). Photolyases are found throughout evolution, except in placental mammals. Like UVDE, photolyases also bind UV lesions directly, but the repair mechanism is very different. Photolyases are very specific to UV lesions, with the ascomycete enzymes only binding CPDs (29, 30). The photolyase protein Phr1 repairs UV dimers using photon energy from the blue end of sunlight (in the laboratory, UVA light is used to activate the enzyme). Thus, Phr1-dependent repair is considered to be photoreactivation-dependent repair, whereas the other UV repair mechanisms are known as dark repair. Reports from E. coli and Saccharomyces cerevisiae also suggest a role for Phr1 in NER by guiding the machinery to the site of the UV lesions (31).
A comprehensive SOS-like response to UV has never been demonstrated in fungi. Nevertheless, some DNA repair/DNA damage tolerance genes have been shown to be induced by UV in S. cerevisiae, Neurospora crassa, and Schizosaccharomyces pombe (32, 33). Another environmental inducer of DNA repair proteins is light. Fungi respond strongly to light, which affects diverse aspects of their biology (34–36). Visible/blue light has been shown to induce UV repair genes, which is correlated with increased repair capacity (29, 37–40). In the fungus Cryptococcus neoformans, uvde has been shown to be regulated by the white collar complex, an important light response determinant. This regulation increases the ability of fungi to survive UVC radiation (37).
All UV repair pathways described here (NER, photolyase, and UVDE) are considered prereplication repair pathways in that they act before the replication fork encounters the UV lesion and thus prevent replication fork arrest by the lesion. Bypassing lesions that escape prereplication repair is often a mutagenic process (41–43). In filamentous fungi, the first nuclear replication during conidial germination is very important, because if a mutation occurs at this stage, it will affect the entire hyphae. Therefore, it would be most beneficial if the first cell division cycles had the highest possible repair capacity.
Due to the soilborne lifestyle of F. oxysporum, we hypothesized that its response to UV irradiation would be inducible. However, using quantitative PCR (qPCR) gene expression analysis, we revealed no clear induction pattern of NER, phr1, or uvde. Instead, we revealed considerable changes in the expression of phr1 and uvde during cell cycle progression and germling development. At first, when germlings contain mainly nuclei in the S phase, the expression of phr1 is induced and that of uvde is reduced. Later, when the germling matures into a hypha and the number of S-phase nuclei decreases, the trend is reversed. We were able to attenuate the expression changes using drugs that block cell cycle progression. Finally, we were able to show that photoreactivation-assisted UV repair is indeed induced when phr1 transcripts are induced. We discuss the meaning of our results in the ecological and evolutionary context of Fusarium wilt disease.
UV repair genes exhibit a complex transcriptional response to UV, visible light, and sunlight.To study the response of F. oxysporum to UV either from a germicidal lamp or the sun, we performed RNA sequencing (RNA-seq) analysis. We used the Lexogen 3′ QuantSeq kit in which only the 3′ end of the gene is sequenced, allowing cost-effective analyses of major trends in gene expression changes across multiple conditions. For accurate determination of the expression of specific genes, we used quantitative PCR, as detailed below. To study the effect of UVC on gene expression, fungi were irradiated at two developmental stages, at 8 and 14 h postinoculation. During 8 h postinoculation, only one or two rounds of replication occur. At 14 h postinoculation, the entire population broke dormancy, and several rounds of the nuclear division had occurred. This is why the analysis of 14 h postinoculation was more detailed. Importantly, from ecological standpoint, 8 h postinoculation resembles the status of the pathogen during early stages in germination and infection, while 14 h postinoculation resembles elongation of the filament and establishment within the root tissue. As described below, the developmental stage of the fungus played a significant role in the response to UV. The different conditions of the RNA-seq analyses are presented in Table 1. The up- and downregulated genes in the different treatments are described in Table S2 in the supplemental material. We analyzed Gene Ontology (GO) terms for over- and underexpressed genes following UV irradiation in both developmental stages. DNA repair was not identified as an enriched module among the overexpressed genes in any of our treatments (Text S1). The GO terms that were enriched following UV exposure are summarized in Text S1. The most significant GO term for upregulated genes at 14 h postinoculation was translation (P = 10−65), and the most significant GO term for downregulated genes at 14 h postinoculation was oxidoreductases (P = 10−4). The most significant GO term for upregulated genes at 8 h postinoculation was transferase activity (P = 10−3), and the most significant GO term for downregulated genes at 8 h postinoculation was RNA processing (P = 10−27). We observed opposite trends in response to UV; GO terms that were upregulated in response to UV when 14 h germlings were irradiated were downregulated when 8 h germlings were treated (Text S1). For example, noncoding RNA processes were upregulated following irradiation at 14 h (P = 10−21) and downregulated following irradiation at 8 h (P = 10−18). As discussed in detail below, we think that this phenomenon is due to the basal uninduced level of expression at the different developmental stages.
RNA-seq analysesa
RNA sequencing results. Fold increases in the expression of genes following UV and sun exposures are shown. The sequencing experiments were done twice, with three different biological replicates in each of them. Download Table S2, XLSX file, 11.2 MB.
Copyright © 2019 Milo-Cochavi et al.
This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.
We next examined the expression of genes that are directly involved in UV DNA damage repair. When 14 h-postinoculation germlings were irradiated with 50 J/m2 UV, an equal number of DNA repair genes were modestly upregulated and downregulated; at 200 J/m2, the expression of UV repair genes decreased (Text S2). The picture was different when 8 h-postinoculation germlings were irradiated. Clear induction of some NER-related genes was observed especially at 50 J/m2, for example, rad1, rad14, rad2, rad4, and xpc (Text S2). Next, we measured the transcriptomic response to 2 h of sunlight exposure at midday (12:00 to 14:00) on a sunny day during the summer in Israel (12 July 2017). The up- and downregulated genes are described in Table S2. GO term analysis did not show significant induction of DNA repair, but it did show induction of genes related to oxidative stress (P = 0.02) (Text S3). Heat map analysis of UV repair genes did not show clear induction by sunlight, and there was a moderate induction of genes related to the circadian clock, such as wc-1 (Text S3).
We measured the expression of individual UV repair genes using quantitative PCR of conidia that were inoculated in potato dextrose broth (PDB) medium either for 8 or 14 h. Several conditions were measured, as follows: (i) UVC, (ii) 10-min exposure to fluorescent light, and (iii) 14 h exposure to solar radiation (Table 1). These analyses clearly showed a change in the expression of several UV repair genes. UV irradiation at 50 J/m2 of 14 h postinoculation germlings reduced the expression of most tested genes, including a 100-fold drop in the expression of uvde (Fig. 1A). Induction by UVC of 5- to 10-fold was observed in uvde, rad14, rad2, and csb when germlings were irradiated at 8 h postinoculation (Fig. 1A). Exposure to visible light resulted in a 10- to 20-fold induction of uvde, rad23, and ddb1 (Fig. 1B). A ca. 10-fold induction of phr1, ddb1, and csb was observed when germlings were exposed to sunlight at 14 h postinoculation (Fig. 1C), in agreement with previous results (9, 40). The effect of UVC on both UV-specific genes phr1 and uvde was highly dependent on the timing of irradiation; we suspected that this was due to the baseline levels of the transcripts at the different time points postinoculation.
Mixed trends in the effects of UV, visible light, and solar radiation on the expression of UV repair genes. Real-time quantitative PCR analysis of the expression of several UV repair genes was performed at 8 and 14 h postinoculation. (A) 50 J/m2 UV followed by 30 min recovery; ef1-a served as a reference gene. (B) Ten minutes of exposure to fluorescent light; gapdh served as a reference gene. (C) Two hours in the sun (see the text and Materials and Methods; gapdh served as a reference gene). In all panels, dark-gray bars indicate 8 h postinoculation, while striped bars indicate 14 h postinoculation.
Oscillations in the expression of UV repair genes during fungal development.The different effects of UV and visible light on gene expression at different time points postinoculation lead us to hypothesize that the cell cycle status of the germling is an important determinant in the basal expression level of UV repair genes. It was previously shown that during the germination and germ tube elongation process of F. oxysporum, only one nucleus is mitotically active, while all the rest are dormant (44). Therefore, while at 8 h postinoculation the ratio between mitotic active and dormant nuclei is expected to be high, at 14 h postinoculation, most of the nuclei are expected to be dormant. Using a histone 1-green fluorescent protein (H1-GFP) fusion encoded by the native locus, we were able to count the number of nuclei at 0, 4, 8, and 14 h postinoculation (Fig. 2A). The populations were significantly different regarding the number of nuclei/germling (P = 2 × 10−16). The number of nuclei/germling at 14 h postinoculation was much higher than at 8 h postinoculation. For example, 47% of the population of 8-h-postinoculated germlings contained only 2 nuclei. We could not detect germlings with 2 nuclei at 14 h postinoculation (Fig. 2A). In contrast, while we could not identify germlings with 6 nuclei or more at 8 h postinoculation, 76% of the 14-h-postinoculation population contained 6 or more nuclei (Fig. 2A). We were also able to show that we could arrest nucleus duplication by using the DNA replication inhibitor hydroxyurea or the mitosis inhibitor benomyl. In both cases, at 8 h postinoculation, the proportion of 2 nuclei germlings from the total population was significantly reduced compared to that with uninterrupted growth (Fig. 2B).
The number of nuclei in germlings 8 and 14 h postinoculation suggests differences in cell cycle profiles. (A) The number of nuclei at 0, 4, 8, and 14 h postinoculation in 160, 100, 100, and 66 germinating conidia, respectively, was determined by analyzing microscope images of a histone H1-GFP strain. The color-coded squares represent the number of nuclei/germlings. (B) Same in panel A, but germinating conidia were treated with 200 mM HU for 4 or 8 h (95 and 75 germlings, respectively) or 50 μg/ml benomyl (BN) for 4 or 8 h (100 germlings).
We hypothesized that the expression of the UV repair genes was altered during the germling development due to the ratio of mitotically active and dormant nuclei. Using quantitative reverse transcription-PCR (RT-qPCR), we measured the expression of phr1 and uvde in untreated conidia from the time of inoculation to 26 h postinoculation. We observed a periodic pattern of expression. In the first 8 h, the expression of phr1 and uvde showed a 5- to 8-fold increase and 16- to 64-fold decrease, respectively; from 8 to 14 h, the trend reversed. From 14 h to 26 h postinoculation, there was another reduction in uvde expression and induction in the expression of phr1 (Fig. 3A). Unlike phr1 and uvde, there was no clear oscillating pattern in the expression of other DNA repair genes such as the repair genes induced by methyl methanesulfonate (MMS) damage (mgt1 and mag1) or NER (rad14, rad2, rad23, and cul4) (Text S4). Some changes in expression were observed for xpc and ddb1, but they were not as pronounced as the changes in uvde and phr1 expression (Text S4).
Opposite oscillations in phr1 and uvde expression correlate the expression of cell cycle genes controlled by cell cycle progression. Quantitative PCR analysis of uvde and phr1 was performed at several time points postinoculation using microconidia in PDB medium. (A) Threshold cycle (CT) values of phr1 and uvde at different time points postinoculation in PDB medium. (B) Ratio of phr1 to uvde expression (as expressed by the inverse ratio of their quantitative PCR CT values) was determined at several time points between 0 and 26 h postinoculation. (C) Changes in the expression of phr1, uvde, and cell cycle-regulated genes (cdc7, cdc28, mcm10, mrc1, and csm3) during the development of germinating conidia. Fold change values were calculated in comparison with the expression in resting conidia.
Cell cycle regulation of oscillations in phr1 and uvde expression.The phenomenon of opposite oscillations in phr1 and uvde expression is interesting because these two enzymes compete for specific binding to the same lesion, but their mechanisms of repair are very different. Whereas Phr1 is only activated during exposure to visible light (including UVA), Uvde can repair UV-induced lesions without light, i.e., after the fungus penetrates the plant tissue or is relocated to the root habitat. Therefore, we calculated the RNA expression ratio of phr1 to uvde (phr1/uvde). In resting conidia, the phr1/uvde ratio was close to 1, while at 8 h postinoculation, it was 100 and returned to a value of 1 at 14 h postinoculation (Fig. 3B).
Developmental biology in fungi is highly dependent on cell cycle progression. At the first 14 h postinoculation, the expression of phr1 showed the same trend as several cell cycle and DNA replication genes such as cdc7, mcm10, and cdc28 (Fig. 3C). In comparison with resting conidia, the expression of these cell cycle genes is increased in the first 8 h; at 14 h postinoculation, the increase is attenuated (Fig. 3C). In contrast, the expression of uvde is decreased in the first 8 h (Fig. 3C). To determine whether the changes in gene expression of uvde and phr1 are controlled by the cell cycle and not merely correlated with it, we measured the expression of these genes under conditions of cell cycle arrest. Two experimental settings were used (Fig. 4A). In the first, conidia were inoculated for 8 h with hydroxyurea (HU) or the M-phase inhibitor benomyl (Fig. 4A). As a control, conidia were inoculated for 8 h without cell cycle inhibition. The ratio between phr1 and uvde was 10 times lower after exposure to 200 mM HU, compared to that with uninterrupted growth (Fig. 4B). HU exposure caused a ca. 10-fold reduction in phr1 expression and changed the amount of uvde only modestly (Fig. 4C). When cells were exposed to 10 μg/ml benomyl, the ratio of phr1 to uvde was also significantly lower than in the control (Fig. 4B) due to a 16-fold increase in the amount of uvde when the benomyl was added (Fig. 4C). The second experimental setting was inoculation of conidia for 8 h in PDB medium and then the addition of either HU or benomyl for another 6 h. As a control, no inhibitor was added. When 200 mM HU was added to the culture 8 h after inoculation, the phr1/uvde ratio was much higher than under uninterrupted growth (Fig. 4B) due to an almost 10-fold increase in the amount of phr1 transcript and a 3-fold reduction in the amount of uvde (Fig. 4C). When 10 μg/ml benomyl was added to the medium 8 h after inoculation, the phr1/uvde ratio was similar to that in the control, but the expression of both genes was reduced by ca. 10-fold (Fig. 4C). For the most part, DNA replication and cell cycle genes responded to HU and benomyl at 8 h postinoculation, similarly to phr1 (Fig. 4D). In conclusion, halting hyphal development through cell cycle arrest disrupted the alterations in the expression of phr1 and uvde.
Opposite oscillations in phr1 and uvde expression are controlled by cell cycle progression. (A) The ratio of phr1 to uvde expression was determined in two experimental scenarios, as follows: (i) 8 h postinoculation in PDB containing 200 mM HU or 10 μg/ml benomyl (BN); and (ii) 14 h postinoculation, where for the first 8 h, conidia were grown in PDB, and then 200 mM HU or 10 μg/ml BN was added for another 6 h. (B) The ratio of the expression of phr1 and uvde following HU and BN exposures as described in panel A. (C) The effect of HU and BN on the expression of phr1 and uvde in the experimental setting described in panel A. The fold increase between treated to untreated was calculated using the ΔΔCT method. For both HU and BN exposures, the normalizing gene was actinL binding protein. (D) The effect of HU and BN on the expression of cell cycle-regulated genes (cdc7, cdc28, mcm10, mrc1, and csm3) during the development of germinating conidia (2 to 8 h postinoculation). Fold increase was calculated using the ΔΔCT method of treated versus untreated.
UV repair efficiency is tightly correlated with oscillations in the expression of UV repair genes.To examine whether the differential expression of phr1 and uvde during fungal development affects DNA repair per se, we measured the repair of cyclopyrimidine dimers (CPDs) facilitated by UVA light. Phr1 binds CPDs in a light-independent manner; however, photolyase activity is dependent on UVA light (photoreactivation). We inoculated conidia for 8 and 14 h, and then the fungi were irradiated with 50 J/m2 UVC; a third of the sample was immediately frozen, a third was kept in water in the dark, and a third was kept in water and irradiated with UVA light. Dark repair (no photoreactivation) was very similar after 8 or 14 h of inoculation (Fig. 5A). The contribution of UVA to UV damage repair was stronger in the 8 h postinoculation than in the 14 h postinoculation group. During the first 2 h of photoreactivation, the repair was twice as efficient when cells were treated at 8 h postinoculation compared to that at 14 h postinoculation (Fig. 5B). We next determined the effect of irradiation timing and photoreactivation on the survival of cells irradiated with different UV doses. Without photoreactivation, there was a small effect of the timing of UVC irradiation on survival, whereas irradiated cells at 8 h postinoculation showed a higher survival rate. However, the major difference was observed under photoreactivation conditions. Photoreactivation improved the survival of irradiated cells at 8 h postinoculation (Fig. 5D). In contrast, for cells irradiated at 14 h postinoculation, survival with and without photoreactivation was almost the same (Fig. 5D). Taken together, the capacity to perform photoreactivation-dependent repair of UV lesions correlated with the pattern of phr1 expression and with higher UV survival.
Photoreactivation-assisted UV repair and survival are enhanced when cells are irradiated 8 h postinoculation than at 14 h postinoculation. (A) A UV repair assay. Conidia were inoculated in PDB for either 8 or 14 h, and then cells were irradiated with 50 J/m2 UVC. After irradiation, cells were incubated at room temperature for the indicated time periods without UVA exposure (see Materials and Methods). Then, the number of UV lesions was determined by immunodot blot assay using antibodies against CPD (see Materials and Methods). The strength of the immunodot blot signal is shown as the percentage of lesions left out of the initial damage after the incubation periods. (B) Same as in panel A, but this time, UVA exposure was done for the indicated time periods. (C) UV survival assay. About 200 to 500 CFU either 8 or 14 h postinoculation were spread on PDA plates and irradiated (0, 50, 150, and 200 J/m2 UVC). Plates were then incubated at room temperature for 1 h with or without UVA exposure (see Materials and Methods). Survival was calculated as the number of CFU after irradiation out of the number of CFU without irradiation.
An adaptation to living in the dark?Fungi have a remarkable ability to respond to visible light. A short illumination period changes the expression of genes for hours afterward. We examined whether the periodic gene expression of phr1 and uvde is changed by exposure to 10 min of a visible light prior to germination. Conidia were harvested in total darkness, exposed to fluorescent light for 10 min, and inoculated in PDB medium for either 4 or 8 h. Remarkably, under these conditions, no oscillations in the expression of phr1 and uvde were observed (Fig. 6). We hypothesize that the oscillations in the expression of UV repair genes are an adaptation to living in the soil, in the dark, despite the fact that F. oxysporum retains the ability to respond to light.
Short illumination period before inoculation abolishes the oscillations in phr1 and uvde expression. Conidia were harvested in total darkness and then illuminated with fluorescent light for 10 min. The conidia were inoculated in PDB for 4, 8, and 14 h. RNA was harvested, and the expression of phr1 and uvde was determined by quantitative PCR. (A) The ratio of phr1 to uvde expression (as expressed by the inverse ratio of their quantitative PCR CT values) was determined at 0, 4, 8, and 14 h postinoculation. Light, 10 min of fluorescent illumination; dark, without illumination. (B) Quantitative PCR CT values of phr1 and uvde at different time points postinoculation in untreated or illuminated conidia. The blue line represents phr1, while the green line represents uvde. Solid lines indicate dark, whereas dashed lines indicate 10 min of light illumination.
The results described here were obtained with microconidia, the most common soilborne spores of F. oxysporum. As described earlier, F. oxysporum has two more types of spores. Chlamydospores are protected from UV by their melanized thick cell wall, but how germlings of chlamydospores react to UV is still unknown. Macroconidia are formed on the infected plants. Whether the results obtained here are relevant to macroconidia is interesting because they can be dispersed by wind (45). We propose a model to put our results in an ecological context. Soilborne microconidia of F. oxysporum f. sp. lycopersici are abruptly exposed to the sun near a tomato plant. Plant signals are perceived by the conidia and stimulate germination toward the tomato plants (13, 14) (Fig. 7). At this stage, exposure to the sun is risky for the survival and genome integrity of the germling, but due to a developmentally regulated program, the expression of phr1 is high, and photoreactivation repair is strong (Fig. 3 and 5). When the germlings further develop, they penetrate the plant tissue. At this stage, the young hyphae are no longer exposed to UV damage on one hand but are also not exposed to visible light on the other hand. Therefore, unrepaired UV lesions cannot be repaired by photolyase. However, at this stage, the expression of uvde is upregulated, and dark repair can occur (Fig. 3, 5, and 7). In nongerminating microconidia that are exposed to the sun, the transcriptional program is changed to a plan that we think fits better an adaptation to light (Fig. 6). This adaptation of F. oxysporum to light yet has to be determined in higher resolution than what is described here (Text S3).
Transcription control of UV repair capacity during the life cycle of F. oxysporum. Shown is a model integrating the expression of UV repair genes in the soilborne fungal plant pathogen F. oxysporum with its life cycle and ecology.
We do not know if the developmentally regulated oscillations in the expression of uvde and phr1 are common to other soilborne fungi. The expression of phr1 in the soilborne fungus Trichoderma harzianum has been shown to be developmentally regulated, because different amounts of the transcript are found in conidiophores, germinating conidia, and mature hyphae (39). However, phr1 expression in T. harzianum was not examined through germination at high resolution, and the same trend may therefore hold in that fungus.
The observed oscillations in gene expression were dependent on cell cycle progression (Fig. 3 and 4). It has been previously shown that developmental stages in fungal plant pathogens are dependent on cell cycle progression. Appressorium formation and host penetration in the rice fungal pathogen Magnaporthe oryzae have been shown to depend on cell cycle progression and, specifically, completion of DNA replication (46). Similarly, in the barley fungal pathogen Blumeria graminis, appressorium morphology is disrupted by HU (47). Interestingly, a connection between the cell cycle, and in particular, DNA replication, and the circadian clock has been previously established in fungi (48, 49). It is possible that inhibition of the cell cycle resets the circadian clock, thus preventing or suppressing the oscillations in phr1 and uvde expression.
In conclusion, we provide novel insight into the regulation of UV repair genes in a soilborne fungus. The demonstrated expression pattern provides a first step in understanding how soilborne fungi can survive abrupt UV exposure while reducing the cost of continuous expression of UV repair genes while in the soil.
Purification and growth of conidia of F. oxysporum f. sp. lycopersici.The starting point of all the experiments described here is resting microconidia (asexual spores) of the fungus F. oxysporum f. sp. lycopersici (strain 4287). Throughout the manuscript, they are termed conidia. To generate conidia for each set of experiments, aliquots from a frozen stock were incubated in KNO3-based medium (1.36 g yeast nitrogen base, 24 g sucrose, and 100 mM KNO3 in 800 ml distilled water) on an orbital shaker (250 rpm) at 28°C for 5 days. Conidia were obtained by filtering the culture through a nylon cell strainer (mesh size, 40 μm; Corning, USA) and resuspended in sterile water usually to a concentration of 1 × 109 to 3 × 109 conidia/ml. This stock of conidia was kept at 4°C for no more than 16 h before the experiments were initiated. This stock also served as time 0 in our experiments. In all of the experiments described here, conidia were inoculated into potato dextrose broth (PDB; BD Sparks, USA) at 0.1 to 1 × 108 conidia/ml. The conidia were incubated on an orbital shaker at 28°C for the indicated time points (as described in the Results section). During the incubation, conidia germinate into germlings that, with time, mature to young hyphae. Finally, the germlings are harvested for either DNA or RNA purification or treated as specified.
UV irradiation and sunlight exposure experiments.Germlings (8 and 14 h postinoculation) were obtained by filtering 10 ml of the cultures that contained 2 × 107 conidia/ml at time 0. The germlings were irradiated with either 50 or 200 J/m2 UV light (254 nm, 15 W lamp; Osram, Germany); the incident fluence was measured with a radiometer (YK-35UV; Digital Instruments, USA). Irradiation was followed by 0, 30, or 60 min of incubation in PDB at 28°C for recovery. To evaluate the impact of sunlight, 14-h-postinoculation germlings were exposed to sunlight for 2 h at 12:00 on a sunny day in the summer in Israel (12 July 2017). Both following UV and sun exposures, the germlings were harvested, immediately flash frozen in liquid N2, and kept at –80°C until RNA extraction was carried out.
RNA extraction.Conidia were disrupted in a Minilys bead beater (Bertin Instruments, France) for 30 s at medium speed. Total RNA was prepared with the Qiagen (USA) RNeasy microkit and was treated on-column with RQ1 RNase-free DNase (Promega, USA) to remove additional residues of genomic DNA. The RNA quality was measured by Agilent 2200 TapeStation machine using an RNA kit (Agilent Technologies, USA). For quantification, 1 μl RNA was measured with a Qubit 2.0 fluorometer and RNA BR kit (Life Technologies, USA).
RNA-seq library construction and next-generation sequencing.cDNA libraries were prepared using 0.5 μg RNA with a QuantSeq 3ʹ mRNA-seq library prep kit FWD (Lexogen GmbH, Austria) and were sequenced using the NextSeq 500 system (Illumina, USA) at the Center for Genomic Technologies of the Hebrew University of Jerusalem. Three biological replicates of each treatment were assessed.
Transcriptome analysis.Raw reads were mapped against the F. oxysporum 4287 (50) reference genome and were counted with HTSeq (51). The reads were assembled to genes, and the gene expression was normalized and quantified using the DESeq2 R package data normalization method (52). Differential expression was assessed for genes expressed in all combinations of UVC treatments versus untreated controls. DESeq2 was used to identify differentially expressed genes based on the negative binomial distribution, adjusted P values (P < 0.05), and log2 fold change values of ≥+1 and ≤−1.
Hierarchical clustering analysis.Heat maps were generated using normalized read count values and the heatmap.2 function of the "gplot" R package (https://www.r-project.org).
Analysis of Gene Ontology terms.Functional enrichment analysis was performed using g:Profiler and one-tailed Fisher's exact test. The g:SCS method was used to compute multiple testing correction with a P value of ≤0.05 as the significance threshold (https://biit.cs.ut.ee/gprofiler/gost).
cDNA synthesis and quantitative PCR expression analysis.The RNA that was purified (see "RNA extraction," above) was also used in a qPCR analysis to determine accurately the relative expression of target genes. cDNA was generated with a FastQuant reverse transcriptase (RT) kit (Tiangen, China), according to the manufacturer's instructions, using 1 μg RNA in a 20-μl reaction mixture at 42°C. Quantitative PCR was carried out using Fast SYBR green master mix (Thermo Fisher Scientific). Each well contained 2 ng cDNA. The final reaction volume in each well was 10 μl in 0.1-ml 96-well plates (USA Scientific, USA). The reactions were carried out on a StepOnePlus real-time PCR system (Thermo Fisher Scientific). Data were analyzed using the StepOnePlus v2.2.3 software (Applied Biosystems). All data were compared using the comparative ΔΔCT method (53). All primer pairs (see Table S1 for a complete list of primers used for all qPCR analyses) amplified their target with equal efficiencies (data not shown).
Primers used in this study. Download Table S1, XLSX file, 0.1 MB.
Cell cycle arrest at different developmental stages.To evaluate the impact of different developmental stages and cell cycle progression on the expression levels of UV repair genes, suspensions of 2 × 107 conidia/ml were subjected to one of two different treatments, as follows: (i) incubation in either 10 ml PDB plus 200 mM hydroxyurea (HU) (Acros; Thermo Fisher Scientific, USA) or 10 μg/ml benomyl (Sigma, Israel) for 8 h, or (ii) incubation in 10 ml PDB for 8 h, followed by the addition of HU or benomyl, as described above, for another 6 h, for a total of 14 h of incubation. Incubations were performed on an orbital shaker (250 rpm) at 28°C. Germlings were harvested, immediately flash frozen in liquid N2, and kept at –80°C, followed by RNA purification and qPCR analysis.
To monitor the number of nuclei per conidia/germling, an F. oxysporum strain that encoded H1-GFP fusion at the native locus was used (44) (kindly provided by Antonio Di Pietro's lab at the University of Córdoba). Images of conidia inoculated in PDB either with or without 200 mM or 50 μg/ml benomyl were taken at 0, 4, 8, and 14 h postinoculation using an SP8 microscope (Leica Microsystems, Wetzlar, Germany). Images were taken using Z stack, laser at 488 nm, with PMT detector range of 497 nm to 550 nm. The objective was HC PL APO CS2 40×/1.10 (water was used for immersion). The refraction index was 1.33 and zoom was 0.75. Statistical significance was determined by applying Pearson's chi-square test for count data using the chi-square.test function of the "MASS" R package (https://www.r-project.org/).
Survival assays.For controlled-environment survival assays, 500 germlings (8 or 14 h postinoculation) were plated on potato dextrose agar (PDA; BD Sparks) medium and were subjected to acute UV (254 nm) treatments (50, 100, 150, and 200 J/m2). Half of the plates were immediately exposed to a UVA white light lamp (15 W; Philips, the Netherlands) at a distance of 25 cm at room temperature for 1 h. After 48 h of incubation at 28°C in total darkness, the surviving colonies were counted.
Measuring repair efficiency of CPDs.Filtered 8- and 14-h germlings were used in 10-ml suspensions of 2 × 107 conidia/ml. Suspensions were irradiated with 50 J/m2 UV light (254 nm); then, half of the samples were subjected to photoreactivation, where spores were exposed to UVA light for 1, 2, 4, 6, and 8 h at a distance of 25 cm at room temperature. Spores were then harvested for analysis of DNA lesion formation and repair, immediately flash frozen in liquid N2, and kept at –80°C.
Immunodot blot assay.Irradiated conidia were disrupted using a Minilys bead beater for 60 s at medium speed. Genomic DNA was purified using 2% (wt/vol) hexadecyltrimethylammonium bromide (CTAB) buffer, followed by a chloroform-isoamyl alcohol (24:1 [vol/vol]) phase separation procedure and precipitation in a final concentration of 50% ice-cold isopropanol. An immunodot blot assay was used to quantify DNA lesions, as described previously (54). Briefly, after denaturation at 95°C for 10 min, 200 ng DNA was combined with an equal volume of 2 M ammonium acetate and placed on ice. Each DNA sample was spotted onto a nitrocellulose membrane (soaked in 6× saline sodium citrate buffer for 10 min at room temperature) in duplicate, using a Minifold dot blot manifold (Schleicher & Schuell, the Netherlands). Membranes were dried in a vacuum gel dryer (model 583; Bio-Rad, USA) for 90 min at 80°C. After blocking in 5% powdered milk in PBST (1 ml Tween 20, 100 ml of 10× phosphate-buffered saline [Biological Industries, Israel], 899 ml double-distilled water), membranes were probed with mouse anti-CPD antibody (NMDND001; Cosmo-Bio, Japan). Following secondary antibody application (peroxidase-conjugated; Jackson ImmunoResearch Laboratories, USA), enhanced chemiluminescence was used to detect the antibody-dependent signal from each DNA spot on-film. The intensity of each spot was quantified using the ImageJ software.
Pretreatment with visible light before inoculation.To evaluate the influence of light exposure before the initiation of conidial germination on the expression levels of UV repair genes at different developmental stages, conidia were obtained by filtering the cultures in total darkness, and suspensions of 10 ml with 2 × 107 conidia/ml were illuminated with fluorescent light (Philips 32-W lamp) for 10 min. Illumination was followed by either 4, 8, or 14 h of incubation in PDB on an orbital shaker at 28°C in total darkness. Germlings were harvested in total darkness, immediately flash frozen in liquid N2, and kept at –80°C, followed by RNA purification and qPCR analysis.
Global analysis of the response to UV radiation in germinating conidia of F. oxysporum. Download Text S1, PDF file, 0.5 MB.
Analysis of the response of UV repair genes to UV radiation in germinating conidia of F. oxysporum. Download Text S2, PDF file, 0.1 MB.
Analysis of the response of UV repair and circadian clock genes to sunlight in germinating conidia of F. oxysporum. Download Text S3, PDF file, 0.1 MB.
NER gene and MMS damage repair gene expression does not oscillate during the germination of F. oxysporum. Download Text S4, PDF file, 0.1 MB.
We declare no conflicts of interest.
This work was funded by the Israel Science Foundation grant 211/16 to S.C.
We thank the lab of Prof. Antonio Di Pietro for providing us an H1-GFP strain.
Received 3 October 2019
Kumagai Y,
Yoshizawa S,
Nakajima Y,
Watanabe M,
Fukunaga T,
Ogura Y,
Hayashi T,
Oshima K,
Hattori M,
Ikeuchi M,
Kogure K,
DeLong EF,
Iwasaki W
. 2018. Solar-panel and parasol strategies shape the proteorhodopsin distribution pattern in marine Flavobacteriia. ISME J 12:1329–1343. doi:10.1038/s41396-018-0058-4.
Albarracín VH,
Pathak GP,
Douki T,
Cadet J,
Borsarelli CD,
Gärtner W,
Farias ME
. 2012. Extremophilic Acinetobacter strains from high-altitude lakes in Argentinean Puna: remarkable UV-B resistance and efficient DNA damage repair. Orig Life Evol Biosph 42:201–221. doi:10.1007/s11084-012-9276-3.
Maslowska KH,
Makiela-Dzbenska K,
Fijalkowska IJ
. 2019. The SOS system: a complex and tightly regulated response to DNA damage. Environ Mol Mutagen 60:368–384. doi:10.1002/em.22267.
Gonzales-Siles L,
Sjöling Å
. 2016. The different ecological niches of enterotoxigenic Escherichia coli. Environ Microbiol 18:741–751. doi:10.1111/1462-2920.13106.
Kreuzer KN
. 2013. DNA damage responses in prokaryotes: regulating gene expression, modulating growth patterns, and manipulating replication forks. Cold Spring Harb Perspect Biol 5:a012674. doi:10.1101/cshperspect.a012674.
Dean R,
Van Kan JAL,
Pretorius ZA,
Hammond-Kosack KE,
Di Pietro A,
Spanu PD,
Rudd JJ,
Dickman M,
Kahmann R,
Ellis J,
Foster GD
. 2012. The top 10 fungal pathogens in molecular plant pathology. Mol Plant Pathol 13:414–430. doi:10.1111/j.1364-3703.2011.00783.x.
Dita M,
Barquero M,
Heck D,
Mizubuti ESG,
Staver CP
. 2018. Fusarium wilt of banana: current knowledge on epidemiology and research needs toward sustainable disease management. Front Plant Sci 9:1468. doi:10.3389/fpls.2018.01468.
Michielse CB,
Rep M
. 2009. Pathogen profile update: Fusarium oxysporum. Mol Plant Pathol 10:311–324. doi:10.1111/j.1364-3703.2009.00538.x.
Alejandre-Durán E,
Roldán-Arjona T,
Ariza RR,
Ruiz-Rubio M
. 2003. The photolyase gene from the plant pathogen Fusarium oxysporum f. sp. lycopersici is induced by visible light and α-tomatine from tomato plant. Fungal Genet Biol 40:159–165. doi:10.1016/S1087-1845(03)00089-6.
Nelson PE
. 1981. Life cycle and epidemiology of Fusarium oxysporum, p 51–80. In Mace ME, Bell AA, Beckham CH (ed), Fusarium wilt diseases of plants. Academic Press, Cambridge, MA.
Iida Y,
Kurata T,
Harimoto Y,
Tsuge T
. 2008. Nitrite reductase gene upregulated during conidiation is involved in macroconidium formation in Fusarium oxysporum. Phytopathology 98:1099–1106. doi:10.1094/PHYTO-98-10-1099.
Ohara T,
. 2004. FoSTUA, encoding a basic helix-loop-helix protein, differentially regulates development of three kinds of asexual spores, macroconidia, microconidia, and chlamydospores, in the fungal plant pathogen Fusarium oxysporum. Eukaryot Cell 3:1412–1422. doi:10.1128/EC.3.6.1412-1422.2004.
Turrà D,
El Ghalid M,
Rossi F,
Di Pietro A
. 2015. Fungal pathogen uses sex pheromone receptor for chemotropic sensing of host plant signals. Nature 527:521–524. doi:10.1038/nature15516.
. 2015. Chemotropic sensing in fungus-plant interactions. Curr Opin Plant Biol 26:135–140. doi:10.1016/j.pbi.2015.07.004.
Agrios GN
. 2005. Plant pathology. Academic Press, St. Louis, MO.
Mao P,
Wyrick JJ,
Roberts SA,
Smerdon MJ
. 2017. UV-induced DNA damage and mutagenesis in chromatin. Photochem Photobiol 93:216–228. doi:10.1111/php.12646.
Ikehata H,
Ono T
. 2011. The mechanisms of UV mutagenesis. J Radiat Res 52:115–125. doi:10.1269/jrr.10175.
Friedberg EC,
Walker GC,
Siede W,
Wood RD,
Schultz RA
. 2006. DNA repair and mutagenesis. ASM Press, Washington, DC.
Goldman GH,
McGuire SL,
Harris SD
. 2002. The DNA damage response in filamentous fungi. Fungal Genet Biol 35:183–195. doi:10.1006/fgbi.2002.1344.
Inoue H
. 2011. Exploring the processes of DNA repair and homologous integration in Neurospora. Mutat Res 728:1–11. doi:10.1016/j.mrrev.2011.06.003.
McCready SJ,
Osman F,
Yasui A
. 2000. Repair of UV damage in the fission yeast Schizosaccharomyces pombe. Mutat Res 451:197–210. doi:10.1016/S0027-5107(00)00050-6.
Lindahl T,
Karran P,
Wood RD
. 1997. DNA excision repair pathways. Curr Opin Genet Dev 7:158–169. doi:10.1016/s0959-437x(97)80124-4.
Moser J,
Volker M,
Kool H,
Alekseev S,
Vrieling H,
Yasui A,
van Zeeland AA,
Mullenders LHF
. 2005. The UV-damaged DNA binding protein mediates efficient targeting of the nucleotide excision repair complex to UV-induced photo lesions. DNA Repair (Amst) 4:571–582. doi:10.1016/j.dnarep.2005.01.001.
Scrima A,
Fischer ES,
Lingaraju GM,
Bohm K,
Cavadini S,
Thoma NH
. 2011. Detecting UV-lesions in the genome: the modular CRL4 ubiquitin ligase does it best! FEBS Lett 585:2818–2825. doi:10.1016/j.febslet.2011.04.064.
Sancar A,
Reardon JT
. 2004. Nucleotide excision repair in E. coli and man. Adv Protein Chem 69:43–71. doi:10.1016/S0065-3233(04)69002-4.
. 1999. Quality control by DNA repair. Science 286:1897–1905. doi:10.1126/science.286.5446.1897.
Kanno S,
Iwai S,
Takao M,
. 1999. Repair of apurinic/apyrimidinic sites by UV damage endonuclease; a repair protein for UV and oxidative damage. Nucleic Acids Res 27:3096–3103. doi:10.1093/nar/27.15.3096.
Yasuhira S,
. 2000. Alternative excision repair pathway of UV-damaged DNA in Schizosaccharomyces pombe operates both in nucleus and in mitochondria. J Biol Chem 275:11824–11828. doi:10.1074/jbc.275.16.11824.
Cervantes-Badillo MG,
Muñoz-Centeno T,
Uresti-Rivera EE,
Argüello-Astorga GR,
Casas-Flores S
. 2013. The Trichoderma atroviride photolyase-encoding gene is transcriptionally regulated by non-canonical light response elements. FEBS J 280:3697–3708. doi:10.1111/febs.12362.
Shimura M,
Ito Y,
Ishii C,
Yajima H,
Linden H,
Harashima T,
. 1999. Characterization of a Neurospora crassa photolyase-deficient mutant generated by repeat induced point mutation of the phr gene. Fungal Genet Biol 28:12–20. doi:10.1006/fgbi.1999.1158.
Sancar GB,
Smith FW
. 1989. Interactions between yeast photolyase and nucleotide excision repair proteins in Saccharomyces cerevisiae and Escherichia coli. Mol Cell Biol 9:4767–4776. doi:10.1128/mcb.9.11.4767.
Davey S,
Nass ML,
Ferrer JV,
Sidik K,
Eisenberger A,
Mitchell DL,
Freyer GA
. 1997. The fission yeast UVDR DNA repair pathway is inducible. Nucleic Acids Res 25:1002–1008. doi:10.1093/nar/25.5.1002.
Fu Y,
Pastushok L,
Xiao W
. 2008. DNA damage-induced gene expression in Saccharomyces cerevisiae. FEMS Microbiol Rev 32:908–926. doi:10.1111/j.1574-6976.2008.00126.x.
Dunlap JC,
Loros JJ,
Colot HV,
Mehra A,
Belden WJ,
Shi M,
Hong CI,
Larrondo LF,
Baker CL,
Chen C-H,
Schwerdtfeger C,
Collopy PD,
Gamsby JJ,
Lambreghts R
. 2007. A circadian clock in Neurospora: how genes and proteins cooperate to produce a sustained, entrainable, and compensated biological oscillator with a period of about a day. Cold Spring Harbor Symp Quant Biol 72:57–68. doi:10.1101/sqb.2007.72.072.
Hevia MA,
Canessa P,
Larrondo LF
. 2016. Circadian clocks and the regulation of virulence in fungi: getting up to speed. Semin Cell Dev Biol 57:147–155. doi:10.1016/j.semcdb.2016.03.021.
Montenegro-Montero A,
. 2015. Around the fungal clock: recent advances in the molecular study of circadian clocks in Neurospora and other fungi. Adv Genet 92:107–184. doi:10.1016/bs.adgen.2015.09.003.
Verma S,
Idnurm A
. 2013. The Uve1 endonuclease is regulated by the white collar complex to protect Cryptococcus neoformans from UV damage. PLoS Genet 9:e1003769. doi:10.1371/journal.pgen.1003769.
Cohrs KC,
Schumacher J
. 2017. The two cryptochrome/photolyase family proteins fulfill distinct roles in DNA photorepair and regulation of conidiation in the gray mold fungus Botrytis cinerea. Appl Environ Microbiol 83:e00812-17. doi:10.1128/aem.00812-17.
Berrocal-Tito GM,
Rosales-Saavedra T,
Herrera-Estrella A,
Horwitz BA
. 2000. Characterization of blue-light and developmental regulation of the photolyase gene phr1 in Trichoderma harzianum. Photochem Photobiol 71:662–668. doi:10.1562/0031-8655(2000)071<0662:COBLAD>2.0.CO;2.
Ruger-Herreros M,
Parra-Rivero O,
Pardo-Medina J,
Romero-Campero FJ,
Limón MC,
Avalos J
. 2019. Comparative transcriptomic analysis unveils interactions between the regulatory CarS protein and light response in Fusarium. BMC Genomics 20:67. doi:10.1186/s12864-019-5430-x.
Boiteux S,
Jinks-Robertson S
. 2013. DNA repair mechanisms and the bypass of DNA damage in Saccharomyces cerevisiae. Genetics 193:1025–1064. doi:10.1534/genetics.112.145219.
Hedglin M,
Benkovic SJ
. 2015. Regulation of Rad6/Rad18 activity during DNA damage tolerance. Annu Rev Biophys 44:207–228. doi:10.1146/annurev-biophys-060414-033841.
Liu B,
Xue Q,
Tang Y,
Cao J,
Guengerich FP,
Zhang H
. 2016. Mechanisms of mutagenesis: DNA replication in the presence of DNA damage. Mutat Res Rev Mutat Res 768:53–67. doi:10.1016/j.mrrev.2016.03.006.
Ruiz-Roldán MC,
Köhli M,
Roncero MIG,
Philippsen P,
Espeso EA
. 2010. Nuclear dynamics during germination, conidiation, and hyphal fusion of Fusarium oxysporum. Eukaryot Cell 9: 1216–1224. doi:10.1128/EC.00040-10.
Rekah Y,
Shtienberg D,
Katan J
. 2000. Disease development following infection of tomato and basil foliage by airborne conidia of the soilborne pathogens Fusarium oxysporum f. sp. radicis-lycopersici and F. oxysporum f. sp. basilici. Phytopathology 90:1322–1329. doi:10.1094/PHYTO.2000.90.12.1322.
Saunders DG,
Aves SJ,
Talbot NJ
. 2010. Cell cycle-mediated regulation of plant infection by the rice blast fungus. Plant Cell 22:497–507. doi:10.1105/tpc.109.072447.
Hansjakob A,
Riederer M,
Hildebrandt U
. 2012. Appressorium morphogenesis and cell cycle progression are linked in the grass powdery mildew fungus Blumeria graminis. Fungal Biol 116:890–901. doi:10.1016/j.funbio.2012.05.006.
Liu X,
Dang Y,
Matsu-Ura T,
He Y,
He Q,
Liu Y
. 2017. DNA replication is required for circadian clock function by regulating rhythmic nucleosome composition. Mol Cell 67:203–213.e4. doi:10.1016/j.molcel.2017.05.029.
Pregueiro AM,
Liu Q,
Loros JJ
. 2006. The Neurospora checkpoint kinase 2: a regulatory link between the circadian and cell cycles. Science 313:644–649. doi:10.1126/science.1121716.
Ma L-J,
van der Does HC,
Borkovich KA,
Coleman JJ,
Daboussi M-J,
Dufresne M,
Freitag M,
Grabherr M,
Henrissat B,
Houterman PM,
Kang S,
Shim W-B,
Woloshuk C,
Xie X,
Xu J-R,
Antoniw J,
Baker SE,
Bluhm BH,
Breakspear A,
Brown DW,
Butchko RAE,
Chapman S,
Coulson R,
Coutinho PM,
Danchin EGJ,
Diener A,
Gale LR,
Gardiner DM,
Goff S,
Hilburn K,
Hua-Van A,
Jonkers W,
Kazan K,
Kodira CD,
Koehrsen M,
Kumar L,
Lee Y-H,
Li L,
Manners JM,
Miranda-Saavedra D,
Mukherjee M,
Park G,
Park J,
Park S-Y,
Proctor RH,
Regev A,
Ruiz-Roldan MC,
Sain D, et al
. 2010. Comparative genomics reveals mobile pathogenicity chromosomes in Fusarium. Nature 464:367–373. doi:10.1038/nature08850.
Anders S,
Pyl PT,
Huber W
. 2015. HTSeq–a Python framework to work with high-throughput sequencing data. Bioinformatics 31:166–169. doi:10.1093/bioinformatics/btu638.
Love MI,
Huber W,
. 2014. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15:550. doi:10.1186/s13059-014-0550-8.
Pfaffl MW
. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45. doi:10.1093/nar/29.9.e45.
Karbaschi M,
Brady NJ,
Evans MD,
Cooke MS
. 2012. Immuno-slot blot assay for detection of UVR-mediated DNA damage. Methods Mol Biol 920:163–175. doi:10.1007/978-1-61779-998-3_12.
You are going to email the following Developmentally Regulated Oscillations in the Expression of UV Repair Genes in a Soilborne Plant Pathogen Dictate UV Repair Efficiency and Survival

redpajama_set_name: RedPajamaCommonCrawl


Today Health Canada announced that the new ACMPR would replace the MMPR on August 24, 2016. This represents the government's response to the decision of the Federal Court in Allard. The new regulations are, in essence, a blend of the MMAR and MMPR.
The Licensed Producers remain the sole commercial option and have become the only source of genetics. This is ironic because those LPs acquired their genetics largely from MMAR producers who now are not allowed to supply each other in the new regime.
Patients will be able to produce for themselves or designated someone to produce for them. The designated producers can supply two patients maximum. Four producers can use a single location. The production amounts will be determined by a formula that appears to mirror the MMAR (5 plants per gram per day indoors, two plants per gram per day outdoor). No language appears to impose maximum dosage or plant limits (at least in the information bulletins). Patients will be able to possess a maximum of 150 grams on their persons.
Dispensaries and pharmacies are not brought into the new system. The government indicates the ACMPR were an immediate solution to Allard and should not be considered long-term.
In my opinion, this represents a significant step forward. The devil is in the details and the full text of the regulations are not currently available. However this response by the Liberal government is significantly more robust and responsive to the Charter ruling than prior responses by the Conservatives.
I will have more updates when the regulations themselves are published.


text:
I blaberidi o scarafaggi giganti (Blaberidae) sono la seconda famiglia più grande dell'ordine dei Blattodea.

Specie notevoli
 Blatta del Capo – Aptera fusca
 Blatta nana di grotta – Blaberus atropos=Blaberus fusca
 Blatta testa di morto – Blaberus craniifer
 Falsa blatta testa di morto – Blaberus discoidalis
 Dubia – Blaptica dubia
 Blatta verde – Panchlora nivea
 Blatta fischiante del Madagascar – Gromphadorhina portentosa
 Blatta dagli occhiali – Nauphoeta cinerea
 Blatta del Suriname – Pycnoscelus surinamensis
 Blatta rinoceronte – Macropanesthia rhinoceros

Generi

 Sottofamiglia Blaberinae
 Archimandrita
 Aspiduchus
 Bionoblatta
 Blaberus
 Blaptica
 Byrsotria
 Eublaberus
 Hemiblabera
 Hiereoblatta
 Hormetica
 Hyporhicnoda
 Lucihormetica
 Minablatta
 Monachoda
 Monastria
 Neorhicnoda
 Oxycercus
 Paradicta
 Parahormetica
 Petasodes
 Phoetalia
 Sibylloblatta
 Styphon
 Sottofamiglia Diplopterinae
 Diploptera
 Sottofamiglia Epilamprinae
 Africalolampra
 Alphelixia
 Anisolampra
 Antioquita
 Apsidopis
 Aptera
 Ataxigamia
 Blepharodera
 Calolampra
 Calolamprodes
 Capucinella
 Cariacasia
 Colapteroblatta
 Dryadoblatta
 Epilampra
 Galiblatta
 Gurneya
 Haanina
 Homalopteryx
 Howintoniella
 Juxtacalolampra
 Litopeltis
 Miroblatta
 Molytria
 Morphna
 Notolampra
 Opisthoplatia
 Orchidoeca
 Phlebonotus
 Phoraspis
 Pinaconota
 Placoblatta
 Poeciloderrhis
 Princisola
 Pseudophoraspis
 Rhabdoblatta
 Rhabdoblattella
 Stictolampra
 Thorax
 Ylangella

 Sottofamiglia Geoscapheinae
 Geoscapheus
 Macropanesthia
 Neogeoscapheus
 Parapanesthia

 Sottofamiglia Gyninae
 Alloblatta
 Gyna
 Paraprincisaria
 Princisaria
 Pseudocalolampra

 Sottofamiglia Oxyhaloinae
 Tribù Gromphadorhini
 Aeluropoda
 Ateloblatta
 Elliptorhina
 Gromphadorhina
 Leozehntnera
 Princisia

 Tribù Nauphoetini
 Griffiniella
 Henschoutedenia
 Jagrehnia
 Nauphoeta
 Rhyparobia
 Simandoa

 Tribù Oxyhaloini
 Oxyhaloa
 Brachynauphoeta
 Coleoblatta
 Heminauphoeta
 Pronauphoeta

 Sottofamiglia Panchlorinae
 Achroblatta
 Anchoblatta
 Biolleya
 Panchlora
 Pelloblatta

 Sottofamiglia Panesthiinae
 Ancaudellia
 Annamoblatta
 Caeparia
 Microdina
 Miopanesthia
 Panesthia
 Salganea

 Sottofamiglia Paranauphoetinae
 Paranauphoeta

 Sottofamiglia Perisphaeriinae
 Bantua
 Compsagis
 Corydidarum
 Cyrtotria
 Derocalymma
 Ellipsica
 Elliptoblatta
 Glomerexis
 Gymnonyx
 Hostilia
 Laxta
 Neolaxta
 Perisphaeria
 Perisphaerus
 Pilema
 Platysilpha
 Poeciloblatta
 Poeciloderrhis
 Pseudoglomeris
 Pseudocalolampra
 Trichoblatta
 Zuluia

 Sottofamiglia Pycnoscelinae
 Proscratea
 Pycnoscelus
 Stilpnoblatta

 Sottofamiglia Zetoborinae
 Alvarengaia
 Capucina
 Lanxoblatta
 Parasphaeria
 Phortioeca
 Phortioecoides
 Schistopeltis
 Schizopilia
 Schultesia
 Thanatophyllum
 Tribonium
 Tribonoidea
 Zetobora
 Zetoborella

 Senza sottofamiglia

 Apotrogia
 Cacoblatta
 Compsolampra
 Cyrtonotula
 Derocardia
 Diplopterina
 Elfridaia
 Eustegasta
 Evea
 Glyptopeltis
 Gynopeltis
 Hedaia
 Isoniscus
 Kemneria
 Mesoblaberus
 Mioblatta
 Paraplecta
 Phenacisma
 Progonogamia
 Pseudogyna
 Pseudoplatia
 Rhicnoda
 Stenoblatta
 Stictomorphna
 Thoracopygia

Note di tassonomia
Tra i generi di sottofamiglia incerta, Eustegasta e Isoniscus sono stati rimossi dalla sottofamiglia Perisphaerinae. Criptocercus è talvolta assegnato a una famiglia separata, i Cryptocercidae.

Note

Altri progetti

Collegamenti esterni

Blattoidei

redpajama_set_name: RedPajamaWikipedia


They say the more you change your hair, the more issues you have. Another way of putting it, which I learned through my mother who read it online, is that people who change their hair a lot do so because they feel it is one of the only things they can control in their lives. I have changed my hair about 16 times, only including hair color switches and one major cut. Oops.
I don't know if these theories are true, but I do know that I love changing my hair. If I could I'd get extensions and then switch to a bob the week after. I've had red, blue, purple, pink, and bleached ends before. I've had red, purple, blonde, brown, and now dark burgundy colored hair. I didn't do anything with my hair last summer so I spent around $200 to get purple hair in the fall…which turned into fried white blonde. So at this point my hair has about 1/3 recovered and my mother is about 50% less mad.
But can I say, "Hair, Voltaire? I'd rather discuss Voltaire" (hello Princess Diaries!). I change my hair for me, for fun, for la joie de vivre! It's cool if you don't like my hair – sometimes I don't like my hair. And that's life yall. Sometimes my hair reflects a change in my life, sometimes it means I'm bored, sometimes it means absolutely nothing. So yeah, last week I cut my own bangs that I don't even wear today. C'est la vie!
Haha, glad you got the reference! Yass girl, change that hair! haha but I support your hair changes! Thanks for reading!
I'm a hair changer too hahaha 🙂 I tried all the colours and styles which left me with a completely damaged hair. I agree with you, sometimes it's really about the change, but mostly (at least for me) it's just fun.


text:
Q: DateInterval failed with decimal number I have a duration in ISO format witch I need to convert it into seconds number. I use the function DateInterval to extract years, mounths, days, hours, minutes and seconds and convert its into seconds. But I have a problem: when the duration in format ISO Contains only int number(PT2M2S), the DateInterval function works perfectly but when  the duration contains decimal number(PT2M2.6S) the DataInterval function does'nt work.
Exemple: $session_time = "PT2M2S";
when I use DataInterval:
`$interval = new DateInterval($session_time);`
$sessionTimeBySeconde = $interval->y*525600*60 + $interval->m*43800*60 + $interval->h*3600 + $interval->i*60 + $interval->s;

In this cas it works perfectly.
Now if I have for exemple : $session_time = "PT2M2.6S";
and if I use DataInterval like above :
`$interval = new DateInterval($session_time);`
$sessionTimeBySeconde = $interval->y*525600*60 + $interval->m*43800*60 + $interval->h*3600 + $interval->i*60 + $interval->s;

In this cas it does'nt work..
For Information I don't responsible for the value $session_time, I juste get it form another system and I want to convert it into seconds.
What I can do ?

redpajama_set_name: RedPajamaStackExchange


Welcome to muzmatch - the Web's favorite place for Bosnian Dating App. Whether you're new to this or finding out about muzmatch for the first time, signup free today and connect with other people from Bosnia and Herzegovina looking for free Muslim Dating App and find your very own muzmatch.
Register with the best Bosnian Muslim dating app and start browsing profiles. It only takes a few seconds.


New Freeware - Free Software Downloads > Home & Education > " Become A Global Home Business Pro -eBook!
15 Free eBooks,11 Free smallbiz Software downloads. Exclusive chapter on Building a Free AD-Machine and get 1000 Hits a day! Free Branding with clickbank ID!
How to Build a FREE-AD Machine that Brings You 1000 Hits a Day! All FREE!
News and Development in Home Business, Freelance opportunities, Internet Marketing and more!
Download " Become A Global Home Business Pro -eBook!


Came on for consideration the above-referenced criminal action, the Court having heretofore referred the request for the revocation of Defendant's supervised release to the United States Magistrate Judge for proper consideration. The Court has received the Report and Recommendation of the United States Magistrate Judge pursuant to its order. Defendant having waived allocution before the Court as well as his right to object to the report of the Magistrate Judge, the Court is of the opinion that the findings and conclusions of the Magistrate Judge are correct.
It is therefore, ORDERED that the Report and Recommendation of United States Magistrate Judge is ADOPTED as the opinion of the Court.
It is further ORDERED that Defendant's supervised release is hereby REVOKED.
It is further ORDERED that Defendant be committed to the custody of the Bureau of Prisons to be imprisoned for a term of thirty-seven (37) months, with a term of supervised release of five (5) years.
The Court also recommends that Defendant be housed in a facility where Defendant Melvin can participate in any combination of psychiatric, psychological, or mental health treatment programs, preferably in the North Texas area, if appropriate.
SIGNED at Beaumont, Texas, this 30th day of November, 2017.


If you want to be part of making art history, you can donate at this link, just use the top donate button for In Context at the top right. space (Sand Point Arts and Cultural Exchange) is acting as our fiscal sponsor so you can deduct your donations as a fully deductible amount. Everything helps honestly, a little goes a long way.


Basics: Body-boosting mousse amplifies hair to give it soft luxurious volume and body.
Benefits: Conditions and moisturizes; Produces unbelievable volume for all hair types.
Bonus: Alcohol and Paraben-free; UV Protectant and Humidity Resistant.