At the Obermann Center, we're in the privileged position of seeing projects through stages from the earliest ambitions to exploratory conversations to what I have come to see as "snapshot" moments—an initial presentation at the Obermann Fellows biweekly seminar, a reimagined syllabus, that first grant application, a sudden meeting of minds at a campus-community conversation, the eureka moment when collaborators connect, the instant students in our graduate institute recognize kindred spirits.
Obermann offers the impetus and the resources for people from across campus to come together to explore and create.
During the 2016–17 academic year, a core group of University of Iowa faculty and graduate students worked to map cultural exchanges across Eurasia from roughly 400 CE  ca. 1450 CE, by focusing on the development, distribution, and sharing of manuscript technologies.
This year's Obermann Humanities Symposium explored Iowa's multicultural history through the largest group of European origin to settle in the state, with a slate of public lectures, exhibits, performances, & conferences.
From book projects to major grant proposals, our Fellows-in-Residence were busy!
The Obermann Center is an active partner in this grant, generously funded by the Andrew W. Mellon Foundation. The grant supports collaboration between the two institutions with a special emphasis on creative teaching with technology.
Paul Dilley, Sarah Bond, and 12 visiting scholars converged to explore and visualize ancient texts in new ways.
Brazilian carnival, network analytics, cochlear implants, and Big Ten diversity programs were all underway at Obermann in Summer 2016.
This year's Institute continued working with the 1855 Johnson County Historic Poor Farm.
Obermann Working Groups provide a shared space for interdisciplinary exploration—an intellectual repast—and a chance to create new intellectual partnerships.
Our funding and people power helped to support scholarly lectures, public art exhibits and performances, workshops, and conference attendance.
A media workshop, a new journal, and several rhetoric seminars filled out POROI's year.
Co-edited by Teresa Mangum, this new book series is a collaboration between the University of Iowa Press and the Obermann Center.
Obermann Conversations seek to bring Obermann scholars into informal dialogue with both a public partner and an interdisciplinary audience.
Last year, the Obermann Center co-sponsored several events that addressed the future of public higher education.
The mission of the DeLTA Center is to discover and communicate the organizing principles of learning and development that are essential for overcoming grand challenges in health and education.
An impressive catalog of the reported publications and projects resulting from Obermann-funded initiatives.
Everyone who helps make the Obermann Center a special space on campus is key to our success!


Two years ago I began an investigation into the breakdown of architecture and the built environment in the event of an apocalypse. Without a definite conclusion in mind, one of the most surprising results of the study has been the importance of architectural sustainability both in the event of an apocalypse, and subsequently to avoid such an event.
In 2010, a competition was launched via the website zombiesafehouse.wordpress.com to design the ultimate zombie safe-house. As noted on the site, applicants were asked to address six programmatic issues one might face in a zombie-ridden world under the premise of a global zombie apocalypse, reducing the world to a 'pre-industrial hostile wilderness'. The first issue is of course to keep zombies out. The five remaining issues, however, are not far off from issues society is facing today including: how to manufacture power, provide water, and dispose of waste.
In a 2011 entry from an unknown applicant, we can see the Eiffel Tower transformed into a self-sustaining home for apocalyptic survivors. A wind turbine has been added to the top of the tower providing power for the inhabitants. A large funnel collects rainwater to be used for drinking and watering crops, which are vertically farmed up the tower's structure. Further entries make use of a wide range of historical monuments around the world ranging from the Statue of Liberty to the Parthenon.
Over the next two weeks negotiators from 195 countries around the world are meeting in Paris at the COP21 (Conference of Parties) also known as the 2015 Paris Climate Conference to develop solutions to this crisis. As climate change is thrust into the global spotlight, so is its impactful relationship with architecture and design. While sustainable architecture gains ground both in terms of necessity and applicability, it is still often viewed as a bonus feature rather than a necessity while buildings today account for more than 40% of energy consumption and roughly a third of all greenhouse gas emissions around the globe.
When we think about sustainable architecture, the term 'energy efficiency' is something that immediately comes to mind; specifically that which corresponds to the daily operation of a particular building. This overarching concern for energy consumption has led to the coining of terms such as 'net-zero' or 'zero-energy' buildings. The idea of 'zero-energy' in this sense is grossly misleading as it ignores the embodied energy (the energy used throughout a building's full life-cycle from construction and maintenance to demolition) and focuses solely on energy used in daily operations.
As we move further into the 21st Century, it is only becoming clearer that a need for architecture that works with nature, rather than against it, is becoming increasingly important. As architects and designers it is our duty to make this change and relay its importance to our clients. Perhaps then it is time for a new definition of architecture; one that moves beyond the traditional notions of form and function, expanding to form, function, and environment.


We would like to introduce you to COURTESY.
If you haven't heard about COURTESY, i will bet you have heard about Dunkel Radio, Apeiron Crew or any of the succesfull projects she has been a part of.
She has archived more this summer than most deejays dare to dream about. With exclusive bookings like Roskilde Festival, Sonar Festival and numerous nightclubs in Berlin.
COURTESY defies the norms of a copenhagen based dj like never seen before. With her soulfull Detroit techno vibe, she truly is a rare commodity.
With musical roots in Dunkel, Culture Box and Jolene she has born a unique groovy deep sound you have to experience.


UniSource is an ultra versitile product that cleans, shines, deodorizes and disinfects with the power of pine oil. After application, a micro coating will protect the surface and reduce scuff marks, gritt and salt errosion. Applying UniSource will restore the floor's original slip resistance.
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Extends the life of existing floor finish significantly!!!
Highly concentrated formula allows for significant cost savings.


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Entertainment Android
The Best Scavenger Hunt Apps and Ideas
Anya Zhukova December 13, 2019 13-12-2019 5 minutes
If you've ever been a part of a well-organized team-building activity, you know that it's a great way to not only have fun together but build trust and understanding among any group of people. That's what makes it such a great way of getting to know each other.
Sure, icebreakers can get people talking, but if you're looking to really make people feel more relaxed with each other, you'll need a proper team-building activity. Like a scavenger hunt. And as usual, there's an app for that.
How to Organize a Scavenger Hunt
So you've decided to organize a scavenger hunt for your family, friends, or colleagues. Don't know where to begin? Worry not, because there are some great scavenger hunt apps available.
Below you'll find our roundup of the best mobile apps and online resources that will do most of the work for you. All you need to do is make sure your friends know where the starting point for the scavenger hunt is.
All scavenger hunt apps have the same basic features. They allow you to create tasks for the teams or individual players to complete. Some of them come with a map, so you can assign your tasks to specific locations. Just make sure your players are equipped with smartphones that have data and GPS enabled during the game.
1. GooseChase
GooseChase is a DIY scavenger hunt platform. This means that you can either choose a theme for your game from the existing "mission bank" on the app, or you can create one from scratch. If you choose to write a unique treasure hunt, you can check out the company's blog for inspiration. You can find things like 15 creative scavenger hunt ideas or 15 funny scavenger hunt ideas.
Once you're happy with the end result, you can send it to the participants' smartphones and begin the hunt.
In order to complete missions, participants submit photos through the app. The tasks vary, and can sometimes involve talking to strangers and even convincing them to take part in the game one way or another. Each mission has a set point value, and the team (or individual) that collects the most points by the end of the game wins.
The game is free if you use it for organizing a small team hunt, and GooseChase offers paid packages for large groups or businesses.
Download: GooseChase for iOS | Android (Free)
2. Locandy
Locandy is another platform that offers ready-made scavenger hunts. More interestingly, they're not just games, but also quests where you have to make decisions. The story changes and evolves depending on what you choose to do.
Locandy offers interactive multimedia games that will help you discover your surroundings while outdoors, or a completely different city while sightseeing. The app and all games are free for users. You need to download them individually before you begin but after that you won't even need an internet connection. Having your GPS enabled is enough.
If you enjoy the Locandy app and scavenger hunts they offer, you can become a contributor. You can request an author account, start creating your own Locandy games, and then share them with your fellow users.
Download: Locandy for iOS | Android (Free)
3. Huntzz
Huntzz is another DIY app that also offers some ready-made treasure hunts. Its main attraction, however, is its antique-looking treasure map interface.
You can create games inside the app, or if you happen to be in the place where other users or the app creators already have shared hunts, you can just pick one of those to play. Most of the hunts are sightseeing-focused, where every clue you get to comes with a so-called "scroll" full of interesting information about a landmark or an exhibit you reached. There's also an option to choose different levels of difficulty, from simple to cryptic game clues.
When you create your own scavenger hunts on the app, you can choose to share them with other users publicly or privately. Some of the games that already exist on the app are free and some you have to pay for. There's also an option of getting full access to all the paid games on the app.
Download: Huntzz for iOS | Android (Free)
4. Scavify
Scavify is a scavenger hunt app that offers complete treasure hunt games for different audiences, such as university students, workplace employees, educational event's attendees, and tourists.
Scavify will offer you different hunts based on various group sizes, whether you want the game to be competitive or not, and the purpose of the scavenger hunt. The tasks in the hunt include taking pictures and videos in specific locations, solving difficult puzzles 5 Puzzle Places for Brain Twisters and Logic Riddles 5 Puzzle Places for Brain Twisters and Logic Riddles If you seek a challenge for your brain, then you're in for a treat. From free printable puzzles to one of the most fiendish video games ever made, we've got it all in store for... Read More , answering quiz-like questions, and scanning QR-codes found in certain places.
One downside might be that Scavify doesn't have a free trial option. Personal packages start at $29 for small groups of 10 players. The company also offers different packages for larger groups and businesses.
Download: Scavify for iOS | Android (Free-to-use, with paid game packages on the site)
5. Google's Emoji Scavenger Hunt
Google's Emoji Scavenger Hunt How to Play Google's Emoji Scavenger Hunt How to Play Google's Emoji Scavenger Hunt AI is usually seen as serious business, but it can be fun too---as evidenced by Google's interesting new Emoji Scavenger Hunt. Read More is a fun little game that you can play using just your phone's camera. After you load the site and start the game, you will only have 20 seconds to complete each task. The tasks are quite simple—you need to find items Google throws at you. A neural network will then try to identify what it is you're holding in front of your camera.
You can even keep the sound on and hear all the items being identified as they come into the camera's view.
If you enjoy this one and feel like playing more fun Google games, check out these quick games you can play on Google Search 8 Secret Google Search Games You Can Play Right Now 8 Secret Google Search Games You Can Play Right Now Did you know Google Search has hidden games you can play? Here are the best ones you can start playing right now. Read More .
6. Geocaching
Geocaching is a scavenger-type activity where users first create "caches" by filling them with items and then hide them wherever they want. They then upload the GPS coordinated to the web for others to find. Once you find a cache, you can replace it with something of equal or greater value and claim what you found.
For an updated map with listings of nearby caches go to the geocaching website or use one of the many (free) geocaching mobile apps. Caches differ based on their size, distance, and difficulty. An easy one could make for a perfect outside-the-box romantic getaway with your significant other.
More Outdoor Games to Play With Smartphones
If you experiment with a few of those apps and tools, you'll definitely crack the formula for your perfect scavenger hunt.
In case these don't quite hit the spot for you, even though you identify as an outdoorsy type who loves exploration and adventure—check out these fun outdoor games to play using your GPS-enabled smartphone 10 Fun Outdoor Games to Play Using GPS-Enabled Smartphones 10 Fun Outdoor Games to Play Using GPS-Enabled Smartphones Using the GPS functionality that's built into all modern smartphones, you can now walk out the door and start exploring the world -- and make a fun game out of it. Read More .
Explore more about: Android Apps, Geocaching, GPS, iOS Apps, .
Radiohead Archives Its Music and Art OnlineDo FM Transmitter Apps Work? How to Transmit Radio From Your Phone
Locatify TurfHunt should definitely be on this list. Easiest way to create Treasure Hunt Games for both indoor and outdoor and with both time-based games and score-based games.
Scavify worked really well for us! By far the best I came across from doing the research of others out there.
Deborah Paterson
That is not how Geocaching works. Do some research first!
Find Your Friends via GPS With These 7 Free Android Apps
The 7 Best Philips Hue Apps for Android and iPhone
The 5 Best Self Defense Apps for Personal Safety When Walking Alone
The 10 Best Workout Apps to Get in Shape
The 5 Best Apps for a Temporary Burner Phone Number
What It Means When Find My iPhone Is Offline and How to Find It Anyway
The Best Camera Apps for Android and iPhone
The 7 Best Free Calling Apps (Unlimited Calls and SMS With Wi-Fi)

redpajama_set_name: RedPajamaCommonCrawl


This entry was posted in Gratitude, Just for Fun, Poetry, Seasons, Winter and tagged chai tea latte, coffee, cold weather, fun, humour, poem, poetry, Tassimo, winter.
🙂 Thanks, Tina! My deepest thanks and respect go out to whoever invented central heating….
Yep – that cold here too. School has been cancelled for tomorrow. And – advisories have been scrolling on the tv all weekend.
Love the comfort of a hot drink in a snowstorm!
Too true! My kids got me a Tassimo for Christmas and it's the best thing to beat the cold right now!!
enjoy your present! I made spiced cider last week and have been enjoying a mug every day.
Guess it proves the gov't cares about some things–that's an encouragement.
Oh my goodness–don't know whether to laugh or cry. Some of the bodies won't show till spring though, right?
*choke* Great gravy and 56/67ths, girl – you DON'T want to know the visual I got on that one….
Brrrrrr….thank goodness for hot drinks and cosy homes.
What a privileged existence we lead – too many blessings to count!
I am flabbergasted by the weather reports and TV news of the snow and cold in the US and Canada.
You should really save poetry like this for the coldest night. There's not even a 4 in the temperature yet!
Wonderful poem in few words, Melody. I can't relate to the cold as our weather has been in the high 60's and sunny. But we're in dire need of rain or we'll be heading into a drought. However, your poem reminds me of a hot pumpkin spice latte, which I indulged in a "few times" over the holiday season. 🙂 Take care and stay warm.


Youth in Arts is a San Rafael-based nonprofit dedicated to providing arts learning for children and teens of all ages and abilities. YIA celebrated its 45th anniversary this year.
I have served on the YIA Board of Directors for the past 9 years. Two of my children participated in 'til Dawn, the award winning youth a cappella group. Both of them continued on to sing in a cappella groups in college, and my daughter Allie is currently a professional singer in Los Angeles. I'm proud of the education YIA provides to a diverse group of students in Marin, and I want to share information about this vibrant and important arts nonprofit with my MCBA colleagues.
Founded to ensure that local children could have ac- cess to music, dance, theater and visual arts—even as funding for school arts programs declined—YIA now reaches over 20,000 students in the North Bay, and has served an astounding one million children over the past 45 years.
Working with an artistically and culturally diverse roster of mentor artists, YIA offers arts programs that link with other classroom curriculum. Although a wealth of research underscores the value of arts education in enhancing student learning, California public schools are consistently underfunded in the arts. A study by SRI International found that two thirds of Marin's principals reported that the lack of funds acted as a barrier in providing arts for their students. YIA programs help bridge this gap.
YIA is a long-time partner with the Kennedy Center's program for in- dividuals with disabilities, providing "Arts Unite Us" residencies tailored for special needs students. This program is the ONLY local arts' program for special education students.
YIA a cappella group 'til Dawn is well known throughout Marin for its talented young singers and its performances at community events like the Marin County Fair, as well as for inspiring audiences at most of Marin's public and private schools. YIA recently founded "C Street Project," a parallel program for advanced teen visual artists. Alumni of these groups have continued to advanced music and arts study, and have successfully launched careers in the arts, thereby building a new creative generation.
Youth in Arts is proud to have served so many of Marin's students for the past 45 years, and looks forward to enriching the lives of children for many more.
For more information about Youth in Arts programs, including current classes and events for youth and families, click on the button below.
Susan Feder is a full time mediator in the areas of personal injury, real estate, employment and business litigation.


text:
Volume & Issue: Volume 13, Issue 1, Winter 2018
1. Prevalence of Clinically Isolated Metallo-beta-lactamase-producing Pseudomonas aeruginosa, Coding Genes, and Possible Risk Factors in Iran
Abdolmajid Ghasemian; Kobra Salimian Rizi; Hassan Rajabi Vardanjani; Farshad Nojoomi
Volume 13, Issue 1 , Winter 2018, Pages 1-9
http://dx.doi.org/10.30699/ijp.13.1.1
Background & Objective: The spread of carbapenem-resistant Pseudomonas aeruginosa is a global concern. Metallo-beta-lactamase (MBL) enzymes cause extensive drug resistance among Gram-negative bacteria. The current study aimed at determining the prevalence of MBL-producing P. aeruginosa in Iran. Methods: ... Read More Background & Objective: The spread of carbapenem-resistant Pseudomonas aeruginosa is a global concern. Metallo-beta-lactamase (MBL) enzymes cause extensive drug resistance among Gram-negative bacteria. The current study aimed at determining the prevalence of MBL-producing P. aeruginosa in Iran. Methods: A total of 43 studies were found out of which 36 were adopted. Data were collected from Google, Google Scholar, Science Direct, PubMed, Scopus, Embase, and Sciverse. The terms "Pseudomonas aeruginosa", "metallo-beta-lactamase", "prevalence", "carbapenems", and "Iran" were searched. Data from the isolates not producing MBLs were excluded from the study. Data were analyzed with Graph Pad Prism 6, meta-analysis section. Results: According to the results of the current study, 36 surveys indicated that 55% of the clinically isolated P. aeruginosa in Iran were resistant to imipenem and meropenem, among which 37.72% were the MBL producers. Among genes encoding MBLs, blaVIM and blaIMP were predominant with the prevalence of 12.91%±11.01% and 12.50%±23.56%, respectively. No report of harboring blaNDM1 and blaSPM1 by P. aeruginosa was found, similar to most of the other countries in Asia. The prevalence of blaVIM and blaIMP from burn settings were 11.50%±3.5% and 24.65%±23%, respectively. Furthermore, the prevalence of these genes was not significantly different among burn and non-burn isolates (P=0.942 and P=0.597, respectively). Moreover, no relationship was observed between the MBL production and patients' age range. Conclusion: Approximately half of P. aeruginosa isolates were carbapenem-resistant in Iran, and approximately half were the MBL producers. The blaVIM and blaIMP were the predominantMBLs among P. aeruginosa strains, while other genes were not found in P. aeruginosa. Moreover, there was no significant difference between blaVIM and blaIMPamong burn and non-burn isolates. Due to the multiple drug resistance conferred by MBLs, detection and control of their spread alongside proper therapeutic regimens in hospitals and community settings are essential to prevent infection acquisition.
2. The Modulatory Effects of Vitamin D on the Expression of IL-12 and TGF-β in the Spinal Cord and Serum of Mice with Experimental Autoimmune Encephalomyelitis
Rayhaneh Ahangar-Parvin; Marzieyeh Mohammadi-Kordkhayli; Sayyed Vahab Azizi; Maryam Nemati; Hossian Khorramdel-Azad; Zahra Taghipour; Zuhair Hassan; Seyed Mohammad Moazzeni; Abdollah Jafarzadeh
Volume 13, Issue 1 , Winter 2018, Pages 10-22
http://dx.doi.org/10.30699/ijp.13.1.10
Background & objective The immunoregulatory effects of transforming growth factor (TGF)-βand interleukin-12 (IL-12) and immunomodulatory actions of vitamin D (VD) were reported in several studies. This study aims to evaluate VD effects on IL-12 and TGF-β expression in experimental autoimmune ... Read More Background & objective The immunoregulatory effects of transforming growth factor (TGF)-βand interleukin-12 (IL-12) and immunomodulatory actions of vitamin D (VD) were reported in several studies. This study aims to evaluate VD effects on IL-12 and TGF-β expression in experimental autoimmune encephalomyelitis (EAE). Methods: EAE was induced in three groups of C57BL/6 mice by immunization with MOG and administered intra-peritoneally 200 ngVD, PBS or olive oil (OO) from day +3 to +30. One group was also considered as healthy control group. At day 31, cytokines expression in the spinal cord and their serum levels were determined using real time-PCR and ELISA, respectively. Results: IL-12 gene expression and its serum levels in PBS-injected- or OO-administrated EAE groups were significantly higher than healthy group. IL-12 gene expression in EAE group treated with VD was significantly decreased compared to PBS-injected- or OO-administrated EAE groups (P Conclusion: VD modulates the expression of IL-12 and TGF-β in spinal cord and serum of EAE mice.
3. Immunohistochemical Characterization of Normal Ovary and Common Epithelial Ovarian Neoplasm with a Monoclonal Antibody to Cytokeratin and Vimentin
Ankita Goel; Nandam Rao; Vissa Santhi; Syam Byna; Bhavana Grandhi; Jyothi Conjeevaram
Background & Objective: The common epithelial ovarian tumors are classified into serous, mucinous, clear cell, endometrioid, the Brenner, mixed, and undifferentiated types. Cytoskeleton intermediate filament composition of ovarian tissues indicates that the cytokeratin and vimentin are observed in ... Read More Background & Objective: The common epithelial ovarian tumors are classified into serous, mucinous, clear cell, endometrioid, the Brenner, mixed, and undifferentiated types. Cytoskeleton intermediate filament composition of ovarian tissues indicates that the cytokeratin and vimentin are observed in ovarian surface epithelium along with the common ovarian epithelial tumors. The current study aimed at investigating the cytokeratin and vimentin expression in epithelial ovarian tumors to establish a diagnostic relevance. Methods: Sixty-six common epithelial ovarian tumors were studied using anti-cytokeratins (Monoclonal Mouse Anti-Human Cytokeratin Clones AE1/AE3; DAKO, Denmark,) and anti-vimentin (Monoclonal Mouse Anti-Vimentin, Clone V9; DAKO, Denmark,) to ascertain the intermediate filament profiles in formalin-fixed and paraffin-embedded surgical pathology materials. Results: All ovarian epithelial tumors expressed cytokeratin in a uniform fashion. Vimentin was coexpressed with high intensity in 62.5% of serous carcinomas, mild intensity in 25% of mucinous adenocarcinoma, and moderate intensity in single case of endometrioid adenocarcinoma. Vimentin decoration in mucinous carcinoma had a focal involvement, whereas malignant endometrioid and serous decoration tended to involve larger areas. There was a significantly increased expression of vimentin in serous cystadenoma and serous carcinoma, compared with their mucinous counterparts. Also, vimentin expression and histologic grade of serous tumors showed a positive correlation. No association was found between vimentin expression and degree of differentiation in mucinous, endometrioid, and Brenner tumors. Conclusion: The current investigation emphasized the efficiency of immunohistochemistry (IHC) typing as a tool for a more precise characterization of the origin and differentiation of human neoplasms.
4. Micronucleus Assay in Exfoliated Buccal Epithelial Cells Using Liquid Based Cytology Preparations in Building Construction Workers
ARUL P; Shetty Smitha; Suresh Masilamani; Akshatha C
Background and objective:Cytogenetic damage in exfoliated buccal epithelial cells due to environmental and occupational exposure is often monitored by micronucleus (MN) assay using liquid based cytology (LBC) preparations. This study was performed to evaluate MN in exfoliated buccal epithelial cells ... Read More Background and objective:Cytogenetic damage in exfoliated buccal epithelial cells due to environmental and occupational exposure is often monitored by micronucleus (MN) assay using liquid based cytology (LBC) preparations. This study was performed to evaluate MN in exfoliated buccal epithelial cells of building construction workers using LBC preparations. Material and methods: LBC preparations of exfoliated buccal epithelial cells from 100 subjects [50 building construction workers (cases) and 50 administrative staffs (controls)] was evaluated by May-Grunwald Giemsa, Hematoxylin and Eosin and Papanicolaou stains. Student's t test was used for statistical analysis and a P value of Results: The mean frequencies of MN for cases were significantly higher than controls regardless of staining methods used.There were statistically significant differences between smokers and non-smokers of the controls as well as duration of working exposure (5 years) and smokers and non-smokers of cases (P=0.001). However, there were meaningful differences regarding mean frequencies of MN between smokers, non-smokers, those with alcohol consumption or not in cases and controls using various stains (P=0.001). Conclusion: There was an increased risk of cytogenetic damage in building construction workers. However, evaluation of MN of exfoliated buccal epithelial cells in building construction workers serve as a minimally invasive biomarker for cytogenetic damage. LBC preparations can be applied for MN assay as it improves the quality of smears and cell morphology, decreases the confounding factors and reduces false positive results.
Original Research Microbiology
5. The Evaluation of Diagnostic and Predictive Values of Helicobacter pylori Stool Antigen Test in Iranian Patients with Dyspepsia
Fahimeh Safarnezhad Tameshkel; Mohammad Hadi Karbalaie Niya; Zahedin Kheyri; Davood Azizi; Farzin Roozafzai; Samaneh Khorrami
Background and Objectives: Iran, as a developing country, is experiencing high burdens of Helicobacter pylori (Hp)-associated non-communicable diseases. Hp stool antigen test (HpSA) is widely used as an inexpensive and feasible noninvasive method to diagnose Hp infection, instead of invasive approaches. ... Read More Background and Objectives: Iran, as a developing country, is experiencing high burdens of Helicobacter pylori (Hp)-associated non-communicable diseases. Hp stool antigen test (HpSA) is widely used as an inexpensive and feasible noninvasive method to diagnose Hp infection, instead of invasive approaches. The current study aimed at evaluating the diagnostic and predictive values of HpSA test for Hp infection in Iranian patients with dyspepsia. Materials and methods: The current cross sectional study was performed on 100 patients with dyspepsia. Gastric mucosal specimens were taken, processed, and examined according to the standard protocols. Simultaneously, stool samples were obtained and sent to laboratory for further analyses. Hp stool antigen titers were assessed using enzyme-linked immunosorbent assay (ELISA) technique. Results:Stool antigen titers were not associated with gender (P-value=0.284), but correlated to age (r=0.213, P-value=0.034).Considering0.385 as a cutoff point, the HpSA test had 80.4% sensitivity and 85.7% specificity. Conclusion: Based on cost-effectiveness of HpSA test, the current study findings corroborated the use of HpSA test to detect and follow-up patients with Hp infection, as an alternative method to detect Hp rather than invasive procedures.
6. Expression of Prostate-Specific Membrane Antigen (PSMA) in Brain Glioma and its Correlation with Tumor Grade
Hiva Saffar; Maryam Noohi; Seyed Mohammad Tavangar; Hana Saffar; Sima Azimi
Background & Objective Angiogenesis is an essential component of tumor growth. Expression of PSMA on the neo-vasculature of many solid tumors, including glioblastoma multi-form, has been determined. The pattern of expression suggests that PSMA may play a functional role in angiogenesis. Methods: ... Read More Background & Objective Angiogenesis is an essential component of tumor growth. Expression of PSMA on the neo-vasculature of many solid tumors, including glioblastoma multi-form, has been determined. The pattern of expression suggests that PSMA may play a functional role in angiogenesis. Methods: expression of PSMA in different grades of brain glioma was evaluated by the immunohistochemistry method to determine the probable usefulness of anti-PSMA antibody as complementary target therapy in different grades of glioma. Results: Overall, 72 cases of low (grade I and II) and high (grade III and IV) grade gliomas were evaluated for expression of PSMA. Positive PSMA staining was observed in 12 (33.3%) of high grade and 3 (8.3%) of low grade gliomas. Although, high grade tumors more commonly had positive result for PSMA (P value=0.009), the intensity of staining was significantly stronger in low-grade tumors (P value=0.009). Conclusion: Expression of PSMA in different grades of glioma might provide a basis for further investigations focusing on selective target therapy in combination with the current standard care in all glioma grades, to improve treatment efficacy.
7. Development of Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Detection of Influenza A, B and Adenoviruses
Mohsen Nakhaie; Hoorieh Soleimanjahi; Hamid Reza Mollaie; Seyed Mohamad Ali Arabzadeh
Background and objective:Millions of people in developing countries lose their lives due to acute respiratory infections, such as Influenza A & B and Adeno viruses. Given the importance of rapid identification of the virus, in this study the researchers attempted to design a method that enables detection ... Read More Background and objective:Millions of people in developing countries lose their lives due to acute respiratory infections, such as Influenza A & B and Adeno viruses. Given the importance of rapid identification of the virus, in this study the researchers attempted to design a method that enables detection of influenza A, B, and adenoviruses, quickly and simultaneously. The Multiplex RT PCR method was the preferred method for the detection of influenza A, B, and adenoviruses in clinical specimens because it is rapid, sensitive, specific, and more cost-effective than alternative methods Methods:After collecting samples from patients with respiratory disease, virus genome was extracted, then Monoplex PCR was used on positive samples and Multiplex RT-PCR on clinical specimens. Finally, by comparing the bands of these samples, the type of virus in the clinical samples was determined. Results:Performing Multiplex RT-PCR on 50 samples of respiratory tract led to following results; flu A: 12.5%, fluB: 50%, adeno: 27.5%, negative: 7.5%, and 2.5% contamination. Conclusion: Reverse transcription-multiplex Polymerase Chain Reaction (PCR) technique, a rapid diagnostic tool, has potential for high-throughput testing. This method has a significant advantage, which provides simultaneous amplification of numerous viruses in a single reaction. This study concentrates on multiplex molecular technologies and their clinical application for the detection and quantification of respiratory pathogens. The improvement in diagnostic testing for viral respiratory pathogens effects patient management, and leads to more cost-effective delivery of care. It limits unnecessary antibiotic use and improves clinical management by use of suitable treatment.
Original Research Biochemistry
8. The Association between Ocular Problems and Serum Testosterone, Prolactin and Thyroglobulin Concentrations in Delayed Phase of Sulfur Mustard Exposure
Hassan Ghasemi; Nayere Askari; Sakine Moaiedmohseni; Soghrat Faghihzadeh; Susan Kabudanian Ardestani; Elham Faghihzadeh; Tooba Ghazanfari
Background and objective Aside from direct toxic effects, Sulfur Mustard (SM) induced serum hormone abnormalities may aggravate ocular complaints, including Ocular Surface Discomfort (OSD) (burning, itching, and redness), dry eye sensation, photophobia, blurred vision, foreign body sensation, and pain. ... Read More Background and objective Aside from direct toxic effects, Sulfur Mustard (SM) induced serum hormone abnormalities may aggravate ocular complaints, including Ocular Surface Discomfort (OSD) (burning, itching, and redness), dry eye sensation, photophobia, blurred vision, foreign body sensation, and pain. The aim of the current study was to investigate the possible association of ocular complaints with serum hormone concentrations in chronic phase of Sulfur Mustard (SM) exposure. Methods:As a part of Sardasht Iran Cohort Study (SICS), 372 SM-exposed patients and 128 non-exposed participants were enrolled. Ocular complaints and ocular surface biomicroscopic conditions and serum hormones were compared. Results:The exposed with tearing group had significantly higher mean serum levels of testosterone and prolactin (ng/mL) than controls (5.75 vs. 4.75, P=0.031; 11.71 vs. 8.42, P=0.009). The exposed with OSD group had significantly higher mean serum levels of prolactin than controls (12.48 vs. 6.90, P=0.002). The exposed with photophobia group had significantly higher mean serum levels of testosterone than the matched exposed (6.25 vs. 5.65, P=0.013). The exposed with blurred vision group had significantly higher mean serum levels of Thyroglobulin (Tg) (ng/mL) than the matched exposed (65.73 vs. 32.6, P=0.003). Conclusion: Higher mean serum levels of testosterone (in exposed with tearing and photophobia) and prolactin (in exposed with tearing and OSD) may play protective roles against SM effects. Higher mean serum levels of Tg may deteriorate the tear film integrity and optical surface, which causes blurred vision. In the chronic phase of SM toxicity, some ocular surface problems are associated with alterations in the serum concentrations of testosterone, prolactin, and Tg.
Original Research Hematopathology
9. Prognostic Significance of Reduction in Ki67 Index After Neoadjuvant Chemotherapy in Patients With Breast Cancer in Kerman Between 2009 And 2014
Vahid Moazed; Elham Jafari; Behjat Kalantari khandani; Ali Nemati; seyedamir benrazavi
Background and objective:Breast cancer is the most common malignancy among women. The Neoadjuvant chemotherapy is the treatment of choice for non-operable tumors. The Ki67 is a proliferation marker that can be used to predict the therapeutic response to chemotherapy and the patients' prognosis. Methods: ... Read More Background and objective:Breast cancer is the most common malignancy among women. The Neoadjuvant chemotherapy is the treatment of choice for non-operable tumors. The Ki67 is a proliferation marker that can be used to predict the therapeutic response to chemotherapy and the patients' prognosis. Methods: This retrospective study was carried out on 55 consecutive patients with breast cancer referred to a Training Tertiary Healthcare Center in Kerman, Iran since 2009 to 2014. After diagnostic approval, the tissue samples of patients were examined for estrogen and progesterone receptors, ki67 and HER2-neu markers by using immunohistochemical staining. Then the patients were treated with 6 cycles of Neoadjuvant chemotherapy regimens by Doxorubicin and Taxans or 4 chemotherapy cycles, containing Anthracycline and Cyclophosphamide and 4 cycles of Paclitaxel. After mastectomy, their samples were reexamined for ki67 again and classified into three groups (low: ki67<15%), medium (Ki67 = 16-30%) and high (Ki67> 30%). Results: Before chemotherapy, 54.5% of the patients had high expression of Ki67. But after chemotherapy, 52.7 of the patients had complete therapeutic response showing that the Ki67 level was reduced significantly (P=0.003). Conclusion: Before and after Neoadjuvant chemotherapy, Ki67 measurements may be used as a predictive marker of therapeutic response.
Short Communication Hematopathology
10. Beware of Bone Marrow: Incidental Detection and Primary Diagnosis of Solid Tumours in Bone Marrow Aspiration and Biopsies; A Study of 22 Cases
Ruchita Tyagi; Aminder Singh; Bhavna Garg; Neena Sood
Background & objective: Introduction: First detection of any solid tumour as metastatic deposits in bone marrow directs clinicians to start searching for the primary tumour. Detection of bone marrow metastasis determines the stage of the malignancy, prognosis, mode of treatment, chemotherapeutic ... Read More Background & objective: Introduction: First detection of any solid tumour as metastatic deposits in bone marrow directs clinicians to start searching for the primary tumour. Detection of bone marrow metastasis determines the stage of the malignancy, prognosis, mode of treatment, chemotherapeutic response and follow-up in case of relapse. The aim of the current study was to analyse the clinico-haematological presentation and morphological pattern of infiltration of solid tumours detected first as metastatic deposits on bone marrow examination. Methods:Three-year retrospective analysis of MGG-stained bone marrow aspiration smears and touch imprints of the bone marrow biopsy and Hematoxylin and Eosin (H&E) stained histopathological sections of biopsies was performed at the Department of Pathology at a tertiary care institute (January 2014 to December 2016). The morphological pattern of metastatic deposits and haematological profiles of the patients were analysed. Exclusion criterion was the presence of hematolymphoid malignancies. Results: In 22 cases, bone marrow was the first site of detection of metastasis. The age of the patients ranged from 3 years and 10 months to 82 years, with equal gender predilection. Overall, 16 cases had cytopenias, 9 cases each had leucocytosis and leukoerythroblastic presentation. The metastasis was from Ewing's sarcoma, prostate carcinoma, gastric adenocarcinoma invasive duct carcinoma breast, gallbladder carcinoma, lacrimal duct carcinoma and invasive papillary urothelial carcinoma. Conclusions: Bone marrow examination is a cheap and reliable investigation to detect metastasis in an unsuspecting case. Bilateral trephine biopsies are recommended to increase the efficacy of detecting bone marrow metastasis.
11. Sinonasal Teratocarcinosarcoma- A Rare Tumour not so Rarely Misdiagnosed
Jayalakshmy Leelamma; Bhavya P. Mohan; Aparna Srinivasan
Sinonasal teratocarcinosarcoma (STCS) is a rare, morphologically heterogeneous and highly aggressive tumour of ambiguous origin. It is characterized by the presence of benign and malignant epithelial, mesenchymal and neuroectodermal components. Because of their rarity and heterogeneity, these lesions ... Read More Sinonasal teratocarcinosarcoma (STCS) is a rare, morphologically heterogeneous and highly aggressive tumour of ambiguous origin. It is characterized by the presence of benign and malignant epithelial, mesenchymal and neuroectodermal components. Because of their rarity and heterogeneity, these lesions are often misdiagnosed, leading to management difficulties. Adequate sampling with a high index of suspicion is needed to diagnose this rare tumour. We reported here a 48-year old man with right nasoethmoidal mass eroding the cribriform plate with intracranial extension. An initial incisional biopsy was performed and a diagnosis of craniopharyngioma was made. Subtotal endoscopic excision of the mass revealed features of STCS. Immunohistochemistry confirmed the same. The patient was subsequently treated with radiotherapy. The histogenesis, histopathological features, immunohistochemistry findings, clinical features and treatment were discussed here. Till date, there are less than 100 cases reported in English literature.
Case Reports Uropathology
12. Sarcomatoid Carcinoma of Renal Pelvis Involving Ureter and Renal Parenchyma with Heterologous Osteosarcomatous Differentiation: A Case Report and Review of Literature
Bhavya P.Mohan; Jayalakshmy PL; V Letha; Suresh Bhat
Sarcomatoid carcinoma is a high‑grade rare malignant tumor with both epithelial and mesenchymal components. Sarcomatoid carcinoma in the upper urinary tract is very rare. We reported here a case of sarcomatoid carcinoma of renal pelvis with osteosarcomatous differentiation, with involvement of the ... Read More Sarcomatoid carcinoma is a high‑grade rare malignant tumor with both epithelial and mesenchymal components. Sarcomatoid carcinoma in the upper urinary tract is very rare. We reported here a case of sarcomatoid carcinoma of renal pelvis with osteosarcomatous differentiation, with involvement of the ureter and renal parenchyma in a 68-year-old female. Histologically, predominant pleomorphic spindle cell sarcoma component with osteoid production and urothelial carcinoma component with in situ areas were identified. Immunohistochemical analysis showed vimentin positivity in sarcomatous component and cytokeratin positivity in carcinomatous component.
Case Reports Gynecologic Pathology
13. Pseudopapillary and Macrofollicular Microscopic Growth Patterns in an Advanced Stage Ovarian Dysgerminoma: A Case Report
Fatemeh Nili; Niusha Noubari; Alireza Abdollahi
ysgerminoma is one of the two most common types of ovarian germ cell tumors. Providing accurate pathologic diagnosis and treatment planning, the prognosis is good even in advanced stages. Pathologic diagnosis is generally straightforward. In microscopic examination, the usual known growth patterns in ... Read More ysgerminoma is one of the two most common types of ovarian germ cell tumors. Providing accurate pathologic diagnosis and treatment planning, the prognosis is good even in advanced stages. Pathologic diagnosis is generally straightforward. In microscopic examination, the usual known growth patterns in tumor cells are solid, trabecular, insular and rarely pseudoglandular. In this paper, we reported an advanced ovarian dysgerminoma with different microscopic patterns of growth, including pseudopapillary and macrofollicular structures, in an 18-year-old woman. The patient underwent staging laparotomy and is currently receiving chemotherapy.
14. Ovarian Small Cell Carcinoma: A Rare Case Report and Review of Literature
Savita Agarwal; Megha Ralli; Pinki Pandey; Shruti Singh
Volume 13, Issue 1 , Winter 2018, Pages 99-102
Ovarian small cell carcinoma is a rare and highly malignant tumor with poor prognosis. It usually presents in younger females with features of hypercalcemia. The exact histogenesis of the tumor is unclear and it may present as an undifferentiated tumor. In such cases, immunohistochemistry (IHC) plays ... Read More Ovarian small cell carcinoma is a rare and highly malignant tumor with poor prognosis. It usually presents in younger females with features of hypercalcemia. The exact histogenesis of the tumor is unclear and it may present as an undifferentiated tumor. In such cases, immunohistochemistry (IHC) plays an important role to confirm the diagnosis. Limited treatment options are available and mainly include radical surgery followed by adjuvant therapy in advanced stages. The current report was a rare case of small cell carcinoma of ovary presenting no symptoms of hypercalcemia diagnosed on histopathological examination and IHC findings.
15. Ectopic Thyroid in Stomach; a Case Report
Parvin Rajabi; Seyed Mehdi Eftekhari; Elham Rouhani; Omid Mirmosayyeb; Azar Baradaran
Volume 13, Issue 1 , Winter 2018, Pages 103-105
http://dx.doi.org/10.30699/ijp.13.1.103
An ectopic thyroid is a form of thyroid dysgenesis in which an entire or parts of the thyroid gland may be located in another part of the body than what is the usual place. The most frequent location is the base of tongue. Although most cases are asymptomatic, symptoms related to tumor size and its relationship ... Read More An ectopic thyroid is a form of thyroid dysgenesis in which an entire or parts of the thyroid gland may be located in another part of the body than what is the usual place. The most frequent location is the base of tongue. Although most cases are asymptomatic, symptoms related to tumor size and its relationship with surrounding tissues, hormonal dysfunction and seldom malignancy may also occur. Any disease affecting the thyroid gland may involve the ectopic thyroid, including malignancy.
16. Reporting an Experience: Improving the Feulgen Staining Technique for Better Visualizing of Nucleus
Noushin Jalayer naderi
Among different staining methods used to demonstrate the nuclear abnormalities, Feulgen is one of the most reliable method. Feulgen staining is specific, sensitive method for evaluating the DNA damages.It has been shown that using non-DNA specific stains for monitoring the nuclear anomalies lead to false-positive ... Read More Among different staining methods used to demonstrate the nuclear abnormalities, Feulgen is one of the most reliable method. Feulgen staining is specific, sensitive method for evaluating the DNA damages.It has been shown that using non-DNA specific stains for monitoring the nuclear anomalies lead to false-positive or false-negative results. From self-experience, immersing the stained slides in hematoxylin for 3-5 minutes in final step after staining the slides with light green, provides a greater ground contrast and very better visualizing of nucleus. By this method, a clear distinction can be made between micronuclei and other structures suggestive of a micronucleus.
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Co-speciation in bedbug Wolbachia parallel the pattern in nematode hosts
Multiple long-range host shifts of major Wolbachia supergroups infecting arthropods
Tiago M. F. F. Gomes, Gabriel L. Wallau & Elgion L. S. Loreto
Feminizing Wolbachia influence microbiota composition in the terrestrial isopod Armadillidium vulgare
Jessica Dittmer & Didier Bouchon
Wolbachia infection dynamics in a natural population of the pear psyllid Cacopsylla pyri (Hemiptera: Psylloidea) across its seasonal generations
Liliya Štarhová Serbina, Domagoj Gajski, … Jessica Dittmer
Using host species traits to understand the Wolbachia infection distribution across terrestrial beetles
Łukasz Kajtoch, Michał Kolasa, … Milada Holecová
Endosymbiont diversity in natural populations of Tetranychus mites is rapidly lost under laboratory conditions
Flore Zélé, Inês Santos, … Sara Magalhães
First insight into microbiome profile of fungivorous thrips Hoplothrips carpathicus (Insecta: Thysanoptera) at different developmental stages: molecular evidence of Wolbachia endosymbiosis
Agnieszka Kaczmarczyk, Halina Kucharczyk, … Sylwia Zielińska
Intracellular symbionts drive sex ratio in the whitefly by facilitating fertilization and provisioning of B vitamins
Yan-Bin Wang, Fei-Rong Ren, … Jun-Bo Luan
Microorganisms in the reproductive tissues of arthropods
Jessamyn I. Perlmutter & Seth R. Bordenstein
Evolutionarily recent dual obligatory symbiosis among adelgids indicates a transition between fungus- and insect-associated lifestyles
Gitta Szabó, Frederik Schulz, … Matthias Horn
Ondřej Balvín ORCID: orcid.org/0000-0003-1816-03711,
Steffen Roth2,
Benoit Talbot3 &
Klaus Reinhardt4
Evolutionary ecology
Wolbachia bacteria, vertically transmitted intracellular endosymbionts, are associated with two major host taxa in which they show strikingly different symbiotic modes. In some taxa of filarial nematodes, where Wolbachia are strictly obligately beneficial to the host, they show complete within- and among-species prevalence as well as co-phylogeny with their hosts. In arthropods, Wolbachia usually are parasitic; if beneficial effects occurs, they can be facultative or obligate, related to host reproduction. In arthropods, the prevalence of Wolbachia varies within and among taxa, and no co-speciation events are known. However, one arthropod species, the common bedbug Cimex lectularius was recently found to be dependent on the provision of biotin and riboflavin by Wolbachia, representing a unique case of Wolbachia providing nutritional and obligate benefits to an arthropod host, perhaps even in a mutualistic manner. Using the presence of presumably functional biotin gene copies, our study demonstrates that the obligate relationship is maintained at least in 10 out of 15 species of the genera Cimex and Paracimex. The remaining five species harboured Wolbachia as well, demonstrating the first known case of 100% prevalence of Wolbachia among higher arthropod taxa. Moreover, we show the predicted co-cladogenesis between Wolbachia and their bedbug hosts, also as the first described case of Wolbachia co-speciation in arthropods.
Interspecific interactions that provide fitness benefits to all partners involved characterizes a large part of the world's biodiversity1,2. One of its prime examples are symbiotic bacteria that are mutualistically connected with their metazoan hosts3. However, the degree of association and type of mutualism may vary among bacteria-metazoan species pairs and depend on environmental and community contexts2. Typical primary symbioses and true mutualisms are characterised by individual bacterial taxa that provide benefits to their host. Benefits to bacteria are rarely measured2 but may be implied if symbionts are restricted to specialised host cells and tissues, and are exclusively transmitted vertically. Secondary symbioses are characterized by more generic, not necessarily host-specific benefits and consequently, bacteria vary in prevalence across cells, tissues and populations of the host. Their effects on a given host may range from pathogenic to mutualistic, and they can be transmitted either vertically, horizontally, or both3.
The mode of symbiont transmission may profoundly affect patterns of phylogenetic co-variation between the symbionts and their hosts. In cases of vertical transmission, symbionts benefit from increased host fecundity, and therefore, selection typically favours mutualistic symbioses4. In contrast, horizontally transmitted symbionts show less dependence on their hosts and even often develop into parasites. Mutually beneficial symbiotic relationships are more likely to lead to congruent lineage divergence in populations of the two partner species than are non-beneficial relationships4,5, in which hosts usually show resistance to parasitic symbionts6. Over evolutionary time scales, the higher co-divergence in beneficial than non-beneficial interacting lineages results in co-speciation, displayed by congruent phylogenies of symbionts and hosts on higher taxonomic levels6. The depth and completeness of the co-cladogenesis pattern has, therefore, been used as a measure of the nature of a particular symbiotic relationship6. In contrast to mutualist relationships, only few cases of co-cladogenesis have been described from parasitic relationships, mainly from ectoparasites (e.g. Hafner's classical gopher study7), but almost none have been reported from single cell parasites living inside host body or cells6.
Wolbachia bacteria are a prominent example of mainly vertically transmitted intracellular endosymbionts. The Wolbachia diversity has been classified into supergroups8, of which up to 17 has been described up to date9. Importantly, across their two major host groups, arthropods and filarial nematodes, Wolbachia vary in whether and how much they benefit their host10. In filarial nematodes, Wolbachia are exclusively mutualistic11 and are characterised as primary symbionts. For example, in different hosts, they are involved in the heme synthesis pathway12 or in ATP provision13, or are crucial for the host's iron metabolism14 and riboflavin provision15. Nematode-associated Wolbachia are usually found in all individuals of a species16,17 and in all species within larger clades9. In these associations, Wolbachia seems to be exclusively vertically transmitted, which is regarded as a sign of host-provided benefits. As predicted, the mutualistic nature between Wolbachia and nematodes usually results in a tight co-speciation9,14,18.
By contrast, in arthropods, Wolbachia are typically parasitic but can provide benefits to hosts4. Wolbachia incidence is reported to be either around 66%19, or 40%20 among arthropod species. Infections often cause host phenotypes with distorted reproduction (reproductive phenotypes, or RPs)10,21. The most common RP is the induction of cytoplasmic incompatibility (CI), where infected females are only able to produce offspring with infected males, and in some cases only with males infected with the same Wolbachia strain10. Other RPs involve the killing or feminization of genetic males22 or the induction of parthenogenesis23. These RPs directly or indirectly increase the proportion of infected females and Wolbachia are, therefore, able to thrive without providing a fitness benefit to the host, despite vertical transmission and dependence on host fecundity.
Theory predicting that fitness benefits to the host may increase symbiont propagation has been confirmed empirically for Wolbachia, even while maintaining the parasitic mechanisms4. For example, the hosts of CI-inducing Wolbachia often display increased fecundity. This leads to a net benefit to the host, if Wolbachia prevail in the population. Following this path, in some arthropods Wolbachia have become an irreplaceable element of the host´s reproduction. For example, Wolbachia controls the apoptosis during oogenesis in Drosophila24 and in the wasp Asobara tabida25, and serves as a sex-determining factor through chromosome formation in the bean borer moth Ostrinia scapularis26. However, despite these sometimes strict dependencies, and unlike in nematodes, there is no evidence for co-cladogenesis of Wolbachia and arthropods20 with a single exception27, which is however ambiguous because horizontal transfer of Wolbachia could not have been ruled out. Indeed, horizontal transmission of Wolbachia among arthropods is frequent28,29,30.
Wolbachia in the common bedbug, Cimex lectularius, are a notable exception to both rules - because they do not cause RP and they are primary symbionts. Hosokawa et al.31 demonstrated that Wolbachia provide biotin and riboflavin, which are essential for bedbug development. In line with an obligatory nature of the relationship, there is a 100% prevalence of Wolbachia in Cimex lectularius populations32. The gene pathway responsible for the synthesis and provision of biotin to the bedbug has been horizontally transmitted from a co-infecting Cardinium or Rickettsia33. The loci in the closely related Cimex japonicus host contained two deletions relative to Cimex lectularius. The authors, Nikoh et al.33, therefore concluded that the biotin production in C. japonicus is dysfunctional, but suggested that its origin lies in a common ancestor of the two Cimex species. In contrast to the laterally acquired biotin genes, the pathway for riboflavin provided to the common bedbug is fully maintained and homologous across Wolbachia in all hosts studied so far34. However, the common bedbug is the only arthropod known to be provisioned by riboflavin produced by Wolbachia34.
Taken together, the characteristics of the bedbug-Wolbachia system are more similar to those of nematodes than other arthropods (primary vs. secondary symbiosis, generic benefits vs. mutualism, complete vs. partial prevalence). Here we test whether these characteristics are reflected in the predicted co-speciation of bedbugs and Wolbachia. If so, it is possible that related bedbug species use a similar type of Wolbachia mutualism based on vitamin provision. We predict that a) a 100% prevalence of Wolbachia within and among bedbug species, b) co-speciation of Wolbachia and their bedbug hosts and c) that across bedbug species, the presence and potential function of the biotin genes shows evidence for a beneficial relationship.
Sampling was restricted to the subfamily Cimicinae and covers all close relatives of the human-associated C. lectularius, a lineage with a known Wolbachia status31,33. The samples comprise the bat-associated lineage of C. lectularius, the sister species C. emarginatus (S. Roth et al., unpublished), and representatives of the three remaining species groups35: C. pilosellus, C. pipistrelli, C. hemipterus, as well as two bird-associated species: C. hirundinis, C. vicarius (formerly classified as Oeciacus – see36). Three species of the closely related genus Paracimex are included as is another bird related Cimex sp. from Japan (Table S1). Specimens were morphologically identified using Usinger's35 and Ueshima's37 keys and compared against an available phylogenetic data base (S. Roth et al., unpublished).
Individuals of each species were collected from as many locations as possible, up to ten, in order to obtain a reliable estimate of intraspecific genetic diversity and a meaningful estimate of prevalence of Wolbachia in bedbug populations. For C. lectularius whose Wolbachia infection status is already known, we analysed one human-associated population and three bat-associated populations. A list of all samples used in this study can be found in Table S1.
DNA analyses
DNA was extracted from longitudinal halves of insect bodies in order to cover all insect tissues and account for all possible bacterial fauna. We extracted DNA for all samples using the DNeasy Blood & Tissue kit (Qiagen) and stored the DNA at −18 °C prior to use. To assess the infection status and reconstruct the phylogeny of the bedbug- specific Wolbachia strain, we studied two Wolbachia loci: the surface protein gene (WSP) and another protein-coding locus (HCPA). Both loci are widely used to characterize infection rate and describe phylogenetic relationships38. To track the beneficial relationship of Wolbachia to bedbug hosts, we chose two biotin loci: BioC and BioH. Each of these was previously found to contain a frameshift in Wolbachia of C. japonicus compared to the fully functional coding sequence in C. lectularius33, suggesting a loss of function in C. japonicus.
For WSP and HCPA loci, we used the universal primers designed for strain typing38. For each of the biotin loci, a pair of primers was designed to cover the whole coding region. For the bedbug phylogeny, we chose two mitochondrial loci, as mtDNA was shown to provide results congruent with a multilocus approach36 and exhibit the same inheritance mode as the Wolbachia infection. For species identification, we used the barcoding fragment of cytochrome oxidase subunit I (658 bp). For specimens chosen for the co-cladogenesis test, we used also a fragment of 16 S (386 bp). All primers used are listed in Table S2.
The target loci were amplified in 50 ul using GoTaq polymerase (Promega), recommended concentrations of other reagents and 2–4 ul of genomic DNA. The annealing temperature for each primer pair is given in Table S2. Purified PCR products were analyzed through a commercial sequencing service (Macrogen Inc. or GATC Biotech).
The HCPA sequences of samples 005 and 069 in C. pipistrelli and 120, 129 and 130 in C. hirundinis showed a secondary signal pointing to an infection by another Wolbachia strain (see Results). We validated by sequencing one sample per species using a newly designed primer HCPA-R2 (see Table S2) which specifically amplified only a single variant.
Phylogenetic analyses
The alignment of the sequence data was carried out using MAFFT39. The phylogenetic analyses were run in IqTree40 and MrBayes 3.041. Each analysis was carried out three times in order to assess convergence. The IqTree was used to infer Maximum Likelihood (ML) phylogenetic trees, using default settings, automatic model choice and 1000 bootstrap alignments. The Bayesian analyses were run using GTR (Generalised Time Reversible) model with gamma-distributed rate variation across sites and a proportion of invariable sites, both with default priors and priors set according to maximum model probability by sampling across GTR model space (lset nst = mixed rates = gamma). The MCMC chain was run in two simultaneous and independent runs for at least 2 million generations in order to achieve a standard deviation of split frequencies below 0.01. For the consensus tree, 10% of trees with unstable probability values were discarded as burn-in.
We chose 7 insect taxa with Wolbachia infections to serve as outgroup samples for phylogenetic analyses, according to a) similarity of the WPS sequence to C. lectularius associated Wolbachia and b) availability of HCPA sequence for the same sample.
The Wolbachia loci WSP and HCPA could have been affected by possible recombination between bacterial strains38. In order to reconstruct the Wolbachia phylogeny and test the co-cladogenesis with bedbug hosts, we used both loci as separate datasets as well as a concatenated dataset and compared the results. The two biotin loci are overlapping regions and were therefore analysed together as a partitioned dataset with independent model parameters for each gene. The two bedbug mitochondrial genes also do not recombine, therefore the same procedure was applied. Among the gene coding sequences, no gene alignment within the bedbugs using the primary sequence signal contained indels, however, the WSP alignment did when outgroups were included. From individuals that showed a secondary signals, we used only the sequence of the primary signal, after it was re-sequenced with the newly designed primer HCPA-R2 (see DNA analyses; Table S2).
In order to test for co-cladogenesis between Wolbachia and the host, we used the Icong index42, a topology-based method, and TreeMap 3.043 which encompasses both distance- and event-based algorithms. In addition, we used Tredist function in Phylip44 to assess the topological Robinson-Foulds distance45 of the mtDNA trees to the Wolbachia trees, comparing to distance to 1000 random trees generated by T-Rex46. For the Wolbachia trees, we used all unique combinations of WSP and HCPA sequences. The sequences of the secondary signal of the C. pipistrelli and C. hirundinis samples were not included (see Results). As the mtDNA variation within bedbug species was greater than that of Wolbachia sequences, we randomly assigned one of the corresponding mtDNA sequences to each unique Wolbachia sequence.
The included co-cladogenesis tests can use only binominal trees. Some of the Bayesian analysis produced polythomies at the terminal tree nodes; we therefore randomly deleted an appropriate number of taxa from the trees to be used. The final count of tree tips is given in Table 1. Using the Icong index, we counted the number of tree tips in the Maximum Agreement SubTree (MAST), and calculated the probability that the Wolbachia and bedbug trees were congruent by chance. The Treemap was used to test the significance of the number of co-divergence events by the Patristic Distance Correlation Test and the significance of the tree congruence was determined by comparing 1000 random Wolbachia trees, taking into account the timing of both bedbug and Wolbachia phylogenies, Priors were set as recommended by the preliminary mapping analysis.
Table 1 Results of co-cladogenesis tests.
Samples identified as Paracimex borneensis and P. setosus appeared likely to be a single species based on sequence data. Since the taxonomy of the genus requires a thorough revision, we retained the identification based on morphology and geography, though the two samples are dealt with as a single species in Table 2.
Table 2 Gene variation for species.
Test of function of biotin loci
We used codon structure integrity of BioC and BioH sequences, and their relative divergence from a common ancestor, to infer the functionality of biotin production of Wolbachia. Paracimex cf. chaeturus and all samples within the clade consisting of Cimex hemipterus, C. pipistrelli group and all bird-related Cimex species provided sequences with two or more overlaying signals. These sequences however clearly represented the same variants differing in length, i.e. deletions were present in some. Most samples within species showed the full-length variant to be clearly dominant and the secondary signal to be of a consistent peak height. Therefore, we could easily separate the full-length sequence from the dominant signal as only two locations were available and the secondary signal was strong, except for in C. vicarius. As an additional safeguard, we manually reviewed the sequence by chromatogram inspection using CodonCode Aligner 6.0.2. The secondary signal was synchronous with the full-length variant until the deletion in the direction of reading. Since all samples were sequenced from both ends of each locus, for each part of the sequence a synchronous and asynchronous double signal was available. The synchronous signal served to detect the variable sites, as well as to correct for within-species variation. The asynchronous signal then served to assess the correct nucleotide at each of the variants. Only two sites in C. pipistrelli, three in Cimex sp. Japan and six in C. japonicus remained ambiguous in the full-length variants.
In order to test whether Wolbachia biotin loci are functional and underlie the symbiotic relationship in bedbug species other than C. lectularius31, we compared all sequences with the sequence of a presumed common ancestor of the sampled species. We reconstructed the ancestral sequences for the biotin loci based on sequences that contained no frame shift or no stop codons, using Mega 7.047. As the node representing the common ancestor we naturally chose the trichotomy between clades consisting of a) C. pilosellus group, b) C. lectularius group and c) a clade consisting of C. pipistrelli group, C. hemipterus and the bird-associated Cimex species (Cimex sp., C. vicarius and C. hirundinis). These three clades correspond to traditional systematics of the genus Cimex and consistently show monophylies in phylogenetic studies, but the relationships among them remain unresolved (Figs 1 and S136, S. Roth et al. unpublished). The divergence in synonymous and non-synonymous changes was computed in PAML 4.848 using the method by Yang & Nielsen49.
The DNA sequences of both bedbugs and Wolbachia used in the study are available at GenBank under codes provided in Table S1.
Wolbachia prevalence
All 68 individuals of all 15 bedbug species tested were Wolbachia-positive (Figs 1 and S1, Table 2). Except for five samples (2/13 from C. pipistrelli, 3/5 from C. hirundinis; see next paragraph), both WSP and HCPA genes showed unambiguous sequences of a single Wolbachia strain present. In those five samples, only HCPA showed a double signal, According to the consistence of the peak heights, this double signal suggests the presence of two Wolbachia strains. The dominant signal belonged to the same Wolbachia strain that was found in other bedbug species. The variation of WSP and HCPA loci of this strain within bedbug species was either zero or very low (Table 2). The low diversity of WSP among the bedbug species allowed for visual inspection of alignment and let us conclude that no codon mismatch or recombination across the WSP sequence had occurred.
Cladograms based on mtDNA (right) and Wolbachia WSP and HCPA loci (left). Bayes posterior probability values are given at each node. For congruence test resuls see Table 1. All unique combinations of sequences of Wolbachia were used, along with mtDNA of corresponding samples. Labels after species name refer to collection site (Table S1). Highlighted are the different positions of a) Cimex adjunctus sample N1, b) C. vicarius and c) C. hemipterus.
The HCPA sequences reconstructed from the secondary signal of the five samples of C. hirundinis and C. pipistrelli pointed to a co-infection by another strain. The sequence and the relative strength of the signal was consistent across specimens of each species, all coming from different geographic locations. It always represented Wolbachia infections belonging to supergroups other than supergroup F according to a clustering with the outgroups (Fig. S1; for sample codes see Table S1). We therefore have solid reasons to believe that the secondary signal represented a single less abundant Wolbachia strain in each Cimex species. Wolbachia supergroups other than F have not been previously found to provide benefits to bedbugs and were, therefore, not tested for co-phylogeny with bedbugs.
Co-cladogenesis
Each dataset (Wolbachia WSP, HCPA, WSP + HCPA, Wolbachia biotin loci, bedbug mtDNA) produced the same topology and very similar posterior probability or bootstrap values across across all runs and priors, either in Bayesian or ML analyses (Figs 1, S1 and S2). The trees based on a particular dataset produced by Bayesian analyses were sometimes less resolved than the ML ones, but otherwise no conflicts in topology were observed. The only topology differences between trees based on different Wolbachia datasets were the varying positions of Cimex hemipterus and C. vicarius.
The bedbug and Wolbachia trees were clearly congruent (Fig. 1). Both Treemap and Icong co-cladogenesis tests unambiguously supported a close congruence of bedbug and Wolbachia phylogenies (Table 1). Among 1000 random trees, the Robinson-Foulds distance of the Wolbachia to the mtDNA trees was always lowest using either Bayesian or ML trees datasets based on any of the Wolbachia datasets.
Across all the six of bedbug mtDNA tree with the Wolbachia trees comparisons (three Wolbachia datasets, two phylogenetic methods), positions of only three taxa differed. Two of them, however, represented unstable topologies across different Wolbachia trees as well (see Discussion).
Biotin function
The Wolbachia biotin loci were successfully amplified in all but two specimens. In one P. borneensis specimen (C94) the BioH primers failed and in P. setosus (C9) neither locus was amplified despite repeated trials. It is not possible to determine whether the failure to amplify was caused by poor DNA quality, the absence of the biotin loci or primer mismatch due to sequence divergence. However, amplification of the HCPA locus using general primers failed as well in these two specimens.
Wolbachia in all samples within the clade consisting of Cimex hemipterus, C. pipistrelli group and the bird-related Cimex species showed at least one additional variant of the biotin loci. The secondary signal was present in Paracimex cf. chaeturus but only visible when the BioC reverse primer was used for sequencing. These variants contained deletions and the pattern of the distribution of the deletions along the genes was largely consistent across species (Table 2). The two deletions in biotin genes found in C. japonicus in a previous study33 were found in the secondary signal of biotin in our samples as well, along with two other deletions.
Biotin sequences drawn from the dominant signal showed low divergence from the presumed ancestral sequence and were similar among most of the bedbug species (Table 2, Fig. S2). No changes in codon structure were detected. In all Paracimex species and in C. hemipterus, BioC sequences contained frameshifts in all variants detected. The divergence from the common ancestor of the sequences drawn from the dominant signal was one order of magnitude higher than in the rest of the species.
Our study revealed two patterns of Wolbachia infection in bedbugs that proved to be unique to arthropods and that have previously been thought to be typical for, and restricted to, nematodes. First, as predicted, we showed a 100% prevalence of Wolbachia within and among the sampled bedbug species. This contrasts with reports from other arthropods showing infection rates that are typically either below 10%, or above 90% within species19,20,50,51, although infections up to 100% have been observed even within presumably parasitic relationships52,53. A high prevalence is predicted in Wolbachia-nematode relationships, where Wolbachia are usually present in all individuals16,17 and where all genera or larger monophyletic clades are infected9.
Secondly, we show, by various methods, the predicted very close co-cladogenesis between bedbugs and Wolbachia, which is a unique finding among arthropods, but is the norm in filarial nematodes and their Wolbachia symbionts9,18. One potential case of co-speciation has been reported in arthropods, namely that between Wolbachia and three Nasonia wasp species27 although horizontal transfer of Wolbachia was not ruled out. Our results show significant congruence between Wolbachia and the bedbug phylogenies in 15 species, regardless of whether any of the Wolbachia loci or the concatenated dataset were used. Three cases of incongruence between bedbug and Wolbachia phylogenies were observed in each analysis (Fig. 1). While such incongruence may be attributable to limited molecular information used in our study, all three cases are also consistent with alternative hypotheses. In one case, the position of Cimex vicarius varied between the Wolbachia and bedbug phylogenies, as well as among different Wolbachia trees. The position of C. vicarius has been very unstable even in previous multilocus phylogeny reconstructions36, possibly due to a conflict between nuclear and mitochondrial molecular data. Wolbachia molecular information is therefore not fully comparable. In the second case, the position of C. hemipterus varied among Wolbachia trees as well. Non-functional biotin provision by Wolbachia and consequently less stringent symbiosis with a host and a different evolutionary divergence are attributable in this case. The third case likely represents a transfer of Wolbachia from C. brevis to the population of C. adjunctus (sample N1, Fig. 1), a possibility facilitated by the fact that the two species are phylogenetically closely related36 and live in geographic vicinity35.
Both results, the complete prevalence and the co-cladogenesis with the host, are predicted4 if, as has previously been found, the bedbug-Wolbachia relationship is based on mutualistic nutritional provision31. The fact that both predictions were upheld simultaneously may indicate that both phenomena are related and may be part of a syndrome characterising the transition from detrimental to beneficial Wolbachia. Two other characters are also correlated with the type of symbiosis and we propose they are part of a syndrome. Nematode-Wolbachia relationships are strictly vertically inherited, whereas horizontal transmission between arthropod species or populations are common5. Finally, nematode individuals or species are infected by single strains, while infections by multiple strains are common in arthropod species. However, despite the striking parallel between the bedbug and the nematode-associated obligate nutrient provision, co-cladogenesis was not entirely congruent between bedbugs and Wolbachia. Five samples, corresponding to two species (C. hirundinis and C. pipistrelli) showed evidence of an infection with two phylogenetically distant Wolbachia strains (Fig. S1). It is currently not clear whether both strains are involved in nutrition provision or represent a case of competition between two strains that would be resolved over time (with the predicted outcome of the mutualistic strain outcompeting the non-mutualistic one4;). The occurrence of two strains also suggests that horizontal transfer of Wolbachia between bedbug species may not be impossible although it is not currently clear how a horizontal transfer might have happened between C. adjunctus and C. brevis (Fig. 1). Sterile interspecific matings have been observed in several bedbug species, including between C. adjunctus and C. brevis35, though sexual transmission of Wolbachia has to our knowledge not previously been observed.
It may also be important to note that while strict co-speciation is considered ubiquitous for Wolbachia-nematode associations14,18, the supergroup F, to which the bedbug-associated Wolbachia belongs31, shows considerable breakdowns of co-speciation patterns with the nematode hosts17,54. In such context, the co-cladogenesis of Wolbachia with bedbugs that we showed may represent a case of the tightest co-phylogeny recorded for the Wolbachia supergroup F.
We found no evidence of an erosion of the protein coding structure in the Wolbachia biotin loci, including in the Wolbachia of C. japonicus. Dissimilar to the study of Nikoh et al.33, we conclude that the Wolbachia biotin synthesis is still in function in most Cimex species (except C. hemipterus, see below). The difference between the two studies may have arisen because of intraspecific variation of Wolbachia in C. japonicus or, alternatively, because our study may have used primers with different specificity than Nikoh et al.33 had.
We observed additional variants of the biotin loci in a monophylum consisting of all bird-associated species, the C. hemipterus and C. pipistrelli species group (i.e. including C. japonicus, see Table 2). These biotin variants contained frameshifts similar to those found in C. japonicus by Nikoh et al.33 and did not represent functional genes. Since functional variants were also present in most of these hosts, the presence of the deleterious ones does not suggest an arrest in biotin provision by Wolbachia to the bedbugs. The deleterious variants are unlikely to originate from co-infecting Wolbachia strains, because WSP and HCPA loci showed no evidence of a secondary infection in the respective specimens, except in the previously mentioned samples of C. hirundinis and C. pipistrelli. However, it is noteworthy that the positions of the deletions in the biotin sequence was very similar among species (Table 2), strongly suggesting a common origin of the deleterious variants within the species clade, though the actual location of the variants remains unknown.
We found deleterious and non-functional sequences exclusively in Cimex hemipterus and in two Paracimex species. The difference between these two groups and the rest of Cimex is further supported by the length of branches of these taxa on the HCPA and WSP trees (Fig. S1), compared to the length of branches in other Cimex species with functional biotin. While most bedbug species in our analysis use biotin provision by Wolbachia, the Wolbachia symbiosis in C. hemipterus and Paracimex spp. is likely evolutionarily different, perhaps dependent on other resources provisioned by Wolbachia, such as riboflavin. This idea may be tested by experimental using Wolbachia manipulations in C. hemipterus and Paracimex.
Regardless of the discussed details, our data provide clear evidence that the syndrome of transition from a host-detrimental to a host-beneficial relationship has evolved in convergence in both bedbugs and filarial nematodes. Both bedbugs and filarial nematode show 100% Wolbachia prevalence and strict co-speciation of Wolbachia and the host taxa. This is exceptional among arthropods, and bedbugs therefore offer a valuable example of evolution of symbiotic relationships. Given the recently proposed possibilities that nematodes that are harmful to humans may be controlled with antibiotics targeting their Wolbachia, it would be interesting to explore whether such a possibility exists in bedbugs, too.
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The authors are thankful for all the collectors that helped gathering the bedbug samples (for names see Table S1). We also thank Dr. Nusha Keyghobadi (Western University, Canada) for English language typographical and grammatical editing. The research was funded by an internal grant of University of Life Sciences, Faculty of Environmental Sciences, no 20164246 IGA FŽP (Ondřej Balvín), and by Meltzer Research Fund (Steffen Roth). Klaus Reinhardt was supported by the Zukunftskonzept of the TU Dresden, funded by the Excellence Initiative of the DFG.
Department of Ecology, Faculty of Environmental Sciences, Czech University of Life Sciences Prague, Kamýcká 129, CZ-165 21, Praha 6, Czech Republic
Ondřej Balvín
The Natural History Collections, University Museum of Bergen, P.O. Box 7800, N-5020, Bergen, Norway
Department of Biology, University of Western Ontario, 1151 Richmond Street, London, N6A 3K7, Ontario, Canada
Benoit Talbot
Technische Universität Dresden, Department of Biology, Applied Zoology, D-01069, Dresden, Germany
Klaus Reinhardt
Ondřej Balvín and Steffen Roth carried out the molecular lab work and participated in data analysis. All authors participated in the collection of material, in the design of the study and in writing the manuscript. All authors gave final approval for publication.
Correspondence to Ondřej Balvín.
Balvín, O., Roth, S., Talbot, B. et al. Co-speciation in bedbug Wolbachia parallel the pattern in nematode hosts. Sci Rep 8, 8797 (2018). https://doi.org/10.1038/s41598-018-25545-y
Accepted: 27 March 2018
DOI: https://doi.org/10.1038/s41598-018-25545-y
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