Enhance patient access to the ideal technology offered to increase their outcomes and top quality of life. So what technologies must we count on to see in the digital wellness market place in 2018? How are these technologies going to influence our lives this year? And what does all this imply for developers of healthcare technology? Study on for our medtech trend forecast for 2018.
Checking health signs such as blood pressure, temperature and mobility generally requires a number of tests and can be time-consuming. A chair created by Sharp is equipped with a number of sensors that can measure a user's important signs all at after and save the data to the cloud for physicians to reference. Sharp developed the chair for patients to use at house and is contemplating adding a videoconferencing technique so patients can go to with physicians remotely. Rather than folks who are ill going to the medical professional, our notion is for wholesome folks to believe about how to keep healthier, prepare for any emergencies and boost their day-to-day way of life," a spokesman said way back in 2013.
The scope of these emerging technologies is breathtaking. High-tech sensors quickly will monitor the at-property cardiac patient's heart each and every minute of each day. A new kind of chip, embedded in a pill will be activated at the precise moment it reaches a patient's stomach, and will confirm for the medical record that he's taking his drugs. Straight out of science fiction, new gizmos will emerge that can scan a physique for a host of symptoms with no poking or prodding and, in seconds, they will make a diagnosis.
Nanotechnology is fulfilling healthcare science's want for more precise treatment options that are much less invasive, much less expensive, and much less difficult to administer than classic methods. That translates into far better patient outcomes, lower healthcare fees, and wider access to healthcare services in beneath-resourced components of the planet.
Our curricular technologies team is made up of Brian Paterson and Brad Miller , who are here to aid students and faculty with computing, learning management computer software, presentations, database software, and other digital wants. EHRs provide invaluable data to clinical researchers, which advances medical expertise and the improvement of new treatments for common health difficulties. In the run-up to national wellness care reform, the very best new medical technologies are designed to make far more people better for less. Right here are a 5 to watch this year.
Not confident what college you want to attend but? has thousands of articles about every single imaginable degree, location of study and career path that can support you locate the school that is proper for you. According to a study from the the University of Michigan, the shift from paper to electronic wellness records reduces the price of outpatient care by three% These researchers estimated this as $five.14 in savings per patient every single month.
There are two primary places of healthcare technology: diagnosis and therapy. And there are literally hundreds of examples of medical technologies we could talk about in every single category. The majority 36 % of the respondents are hugely satisfied with Housing and education facilities.
In this lesson, find out what health-related technologies is and be able to list numerous examples of diagnosis and remedy healthcare technologies, both basic and complicated. Around 270 million Americans own a mobile phone , and even a lot more are connected on the internet. As with any market, healthcare has necessary to transform its communication processes to connect with individuals wherever they are.
We unite Penn's interdisciplinary strengths in Medicine, Engineering, and beyond to generate transformative technologies that address the world's pressing overall health care needs. The following year, our 2017 health sector trend report listed technologies that have been set to effect the industry the most: Electronic Wellness Records, 3D printing, connected wearables, mobile devices, and applications. For medtech developers, modifications in the regulatory atmosphere and information safety concerns were expected to be the top-ranking challenges of 2017.
An MRI scanner, for example, creates photos of the tissues inside your body – even the brain! We do this to check for organ abnormalities, to look for cancerous tissue, to spot torn ligaments, and a lot of other uses. It performs by surrounding a particular person with a large magnet. This aligns the magnetic particles in a person's body. When you take away the magnetic field, the speed at which they return to standard is diverse depending on the variety of tissue, and this can be detected and utilized to develop an image. Other technologies that are utilised to take images incorporate CT scanners, and x-ray machines.
Continuous technological developments in healthcare have saved numerous lives and enhanced the top quality of life for even a lot more. Not only has technologies changed experiences for individuals and their households, but it is also had a huge influence on medical processes and the practices of healthcare professionals.
We launched Field Service in 2002 and have been committed to supporting the development of the service sector ever since. What started off as 100 folks in a area discussing the future of service has turn into 500 senior-level service executives becoming inspired while learning and developing their firm as properly as their careers. At Field Service Medical, whether or not it really is an interactive roundtable discussion, case study presentation, panel, or one-on-a single Q&A, you will have the likelihood to get the options you are looking for. You will acquire targeted, niche studying possibilities throughout the conference. Assured.
We unite Penn's interdisciplinary strengths in Medicine, Engineering, and beyond to generate transformative technologies that address the world's pressing overall health care requirements. In the run-up to national wellness care reform, the best new health-related technologies are designed to make a lot more men and women much better for much less. Here are a five to watch this year. With the shift to EHRs (and the reality that even one research study can quantity to one hundred terabytes of information), healthcare facilities require to have expandable, price-effective, and secure storage options. This is where The Cloud comes in.
The scope of these emerging technologies is breathtaking. Higher-tech sensors soon will monitor the at-home cardiac patient's heart each minute of each and every day. A new type of chip, embedded in a pill will be activated at the precise moment it reaches a patient's stomach, and will confirm for the medical record that he's taking his drugs. Straight out of science fiction, new gizmos will emerge that can scan a body for a host of symptoms with out poking or prodding and, in seconds, they'll make a diagnosis.
Telehealth is improving allied healthcare jobs, including some of the top-paying roles in the field , such as health-related assistants. The implementation of these telemedicine choices signifies less crowded waiting rooms and easing the pressure on front desk teams.
This technology's prospective was significantly demonstrated when a Kaiser patient suffered a tear in the wall of his aorta, the primary artery major out of his heart. The clinical team printed his artery in three-D and really went via several various scenarios on how they could insert a stent to prevent additional rupture," Chai says. They utilized that in a team-based coaching atmosphere to see how they could confidently proceed with some of these unique procedures." Chai compares the method with a flight simulator in which a pilot masters the intricacies of the cockpit before getting into a genuine one. The innovation enables us to create a much more specialized, customized, precise therapy program," Chai explains. Ultimately, that improves the high quality and affordability of care." The patient, by the way, came by way of the process fine and is recovering.


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1862 June 21st
June 21 1862 Saturday
Battle of Simmon's Bluff, SC (CWSAC Limited Battle Union Victory)
Naval Assault on Vicksburg
Kentucky. Incident at Owen County.
Mississippi. Expedition to Hernando. Incident at Coldwater Station.
Mississippi. USS Bohio captured the sloop L Rebecca bound from Biloxi to Mobile.
Simmon's Bluff, South Carolina. The Union 55th Pennsylvania under Lieutenant A C Rhind landed from the gunboat USS Crusader and transport Planter near Simmon's Bluff on Wadmelaw Sound. The amphibious expedition aimed to cut the Charleston & Savannah Railroad. They surprised and burned an encampment of the 16th South Carolina Infantry (Colonel James McCullough). The Confederates scattered but the Union troops returned to their ships and abandoned their raid on the railroad. No casualties were reported. (CWSAC Limited Battle Union Victory)
Tennessee. Incidents at Battle Creek, Shellmound, Rankin's Ferry and Jasper.
Tennessee. Union Major-General Ulysses Simpson Grant departed for Memphis to set up headquarters for the Army of Tennessee, with command over the divisions of Major-General William Tecumseh Sherman and Major-General John Alexander McClernand.
Virginia. Incident at Fair Oaks Station.
USA: The Northern District of the Department of the South was discontinued
USA: The Southern District of the Department of the South was discontinued.
USA: The Western District of the Department of the South was discontinued.
USA: The Sub-District of Jackson was established in the District of West Tennessee, comprising the area around Jackson, City, Bolivar, Grand Junction, La Grange, Trenton and Humboldt, Tennessee.
USA: Major-General John Alexander McClernand assumed command of the Sub-District of Jackson.
McClernand, John Alexander | Kentucky-Illinois | Born 30 May 1812 Hardinsburg, Kentucky | Private USV 1832 Brigadier-General USV August 6 1861 to rank from 17 May 1861 Major-General USV March 22 1862 to rank from March 21 1862 Resigned USV November 30 1864 | Died 20 September 1890 | 21 June 1862 – 24 August 1862 Sub-District of Jackson, 21 December 1862 – 3 January 1863 XIII Corps Tennessee, 4 January 1863 – 11 January 1863 Army of the Mississippi, 31 January 1863 – 18 June 1863 XIII Corps Tennessee, 20 February 1864 – 14 March 1864 XIII Corps Gulf
CSA: The Department of North Carolina was extended to include the south bank of the James River in Virginia, including Drewry's Bluff.
Department No 1: Earl Van Dorn
Western Department: Pierre Gustave Toutant Beauregard interim Braxton Bragg awaited
Army of the West: Earl Van Dorn interim John Porter McCown temporary awaited
Brigadier-General USV Isaac Ferdinand Quinby 20 March 1862 to rank from 17 March 1862 Sub-District of Columbus
Colonel USV Thomas Maley Harris 20 May 1862 Cheat Mountain District
General ACSA Pierre Gustave Toutant Beauregard 31 August 1861 to rank from 21 July 1861 Western Department interim
General PACS Braxton Bragg March 7 1861 Western Department awaited & Army of Mississippi
Major-General PACS Earl Van Dorn 19 September 1861 Department No 1 & District of Southern Mississippi and Eastern Louisiana & Army of the West interim
Major-General PACS John Porter McCown 11 March 1862 to rank from 10 March 1862 Army of the West temporary awaited

redpajama_set_name: RedPajamaCommonCrawl


Baton Rouge, LA – Waitr, the popular restaurant on-demand platform, has announced the formation of the Waitr Restaurant Incubator Lab. The new initiative is designed to accelerate early stage growth of a startup restaurant, providing aspiring chefs the ability to create a brand and design a menu to make their businesses successful. The lab is located in Waitr's just-opened Baton Rouge office.
Waitr is seen as the industry's top food delivery service for customers and businesses, despite launching only three years ago. Founded in Lake Charles, Louisiana, Waitr has quickly grown to serve more than 150 cities across the Southeast, including Baton Rouge. It currently has 3,000 employees and partners with more than 4,000 restaurants nationwide.
Waitr's new Baton Rouge office was formerly occupied by Baton Rouge meal kit company Indie Plate. Additionally, Waitr has acquired select assets from Indie Plate, including a full commercial kitchen to launch the restaurant incubator program. The employees of Indie Plate have all been hired by Waitr and will be integral in working on the program's development, as well as other elements of Waitr's core business.
"We're thrilled to be part of this new enterprise by Waitr," said Peru Sharma. "Everyone on our team has invested many hours in the food delivery business … and with this new venture, the staff is genuinely excited to be a hub for such an innovative idea.
Waitr has received acclaim for distinguishing itself with its distinct technology. The company has been widely lauded for its growth cornerstones, including its live monitoring of deliveries yielding faster door-to-door service, its unique ability to provide full-color photography of every menu item from all participating restaurants and its go-to market strategy.


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St. Bonaventure names Valento vice president for enrollment
Mar 17, 2015 | University president Sr. Margaret Carney, O.S.F., today announced the appointment of Bernard (Bernie) Valento as vice president for enrollment.
Buffalo Center to host grad school information session March 21
Mar 16, 2015 | A graduate school information session will be held from 2-4 p.m. Saturday, March 21, at St. Bonaventure University's Buffalo Center, located at Hilbert College, 5200 South Park Ave.
Nestlé Waters North America CEO Tim Brown to be keynote speaker at St. Bonaventure Commencement
Mar 12, 2015 | Tim Brown, president and chief executive officer of Nestlé Waters North America since 2013, will be the keynote speaker for St. Bonaventure University's 155th Commencement ceremony May 17.
Students urged to attend Fulbright meeting Thursday
Feb 25, 2015 | An informational meeting on the Fulbright U.S. Student Program will be held in Plassmann 108 at 11:30 a.m. Thursday, Feb. 26.The meeting is open to all students and will explain the application and selection process.
SBU's Slavny qualifies for ski nationals in Oregon
Feb 24, 2015 | St. Bonaventure University sophomores and alpine ski racers Gabby Slavny and Max Borgus competed in the USCSA National Championships Mideast Conference Tournament Feb. 21-22 at Bristol Mountain in Canandaigua.
St. Bonaventure's Homecoming Weekend kicks off Feb. 26
Feb 17, 2015 | St. Bonaventure's 41st annual Homecoming Weekend begins Thursday, Feb. 26, and concludes on Sunday, March 1.
Bonnies to rally this weekend with marrow drive to help alumnus stricken with cancer
Feb 04, 2015 | The Student Government Association at St. Bonaventure University will host a Bone Marrow Registry Drive in an effort to help alumnus John Hyland, Class of '89, who is in need of a bone marrow transplant.
Students invited to learn about career opportunities in nonprofit and volunteer organizations
Feb 04, 2015 | St. Bonaventure will host a series of programs in February aimed to offer students inspiration and information about career opportunities in non-profit and volunteer organizations.
Nominate a remarkable SBU individual or organization for a 2015 Doino award
Jan 29, 2015 | Show support of your St. Bonaventure family by nominating one of our remarkable individuals or organizations for recognition at the 2015 Fr. Joe Doino, O.F.M., Honors & Awards Program.
Winter Carnival on Jan. 31 to mark Thomas Merton's 100th birthday
Jan 22, 2015 | St. Bonaventure University will celebrate the 100th anniversary of Thomas Merton's birth with a winter carnival on Saturday, Jan. 31.
St. Bonaventure generated $144 million economic impact in 2013, new study reveals
Jan 16, 2015 | St. Bonaventure University generated a regional economic impact of more than $144 million in 2013, according to analysis released this week by the Commission on Independent Colleges and Universities.
Unique NSF grant program allows St. Bonaventure to offer 25 STEM scholarships for fall 2015
Jan 06, 2015 | The National Science Foundation has awarded St. Bonaventure University almost $600,000 to help recruit talented students interested in science, technology, engineering and math.
St. Bonaventure, JCC co-sponsoring financial aid workshop Jan. 5
Dec 17, 2014 | St. Bonaventure University will host a College Financial Aid Workshop at 6:30 p.m. Monday, Jan. 5, in the Doyle Hall Dining Room.
Zwierlein returning to St. Bonaventure as interim director of athletics
Dec 10, 2014 | Dr. Ron Zwierlein is coming back to the institution he helped revive a decade ago. The former St. Bonaventure University athletic director will assume the same position on an interim basis beginning Dec. 29. Zwierlein replaces Steve Watson, who has accepted the athletic director position at Loyola Chicago after eight years at St. Bonaventure.
Bronx man to be student speaker at SBU's December Recognition
Dec 09, 2014 | Journalism and theater double-major Frederick "Freddie" Alvarez of the Bronx was selected as the student speaker at St. Bonaventure University's December Recognition Ceremony Saturday, Dec. 13.
SBU's AMA chapter fundraiser Dec. 5 gets ugly
Dec 04, 2014 | The American Marketing Association chapter at St. Bonaventure University had to get crafty to fundraise this year.
SBU senior collecting winter coats to aid refugee families in Rochester
Dec 04, 2014 | Western New York is not a winter wonderland for those who aren't accustomed to the harsh weather conditions, so senior Rachael Herrmann is gathering coats of different sizes to donate to refugee families at Saint's Place in Rochester, N.Y.
Bonaventure students provide perspectives and advice for finals week
Dec 03, 2014 | St. Bonaventure University students weighed in on their pre-finals study habits, freshmen adding new perspectives and upperclassmen offering advice to underclassmen.
Holiday exhibitions to deck the halls of St. Bonaventure University's Quick Center
Nov 25, 2014 | Two exhibitions will entertain visitors to the Regina A. Quick Center for the Arts at St. Bonaventure University over the holidays.
St. Bonaventure now offering undergraduate degree in Professional and Creative Writing
Nov 24, 2014 | St. Bonaventure University has unveiled a new undergraduate major that combines the imaginative power of creative writing with the technical skill of professional communicators.


I ran across this video the other day. It was very entertaining, and it reminded me of what it was like to be a kid. Endless adventure. An innate sense of invention, innovation, and creativity. Come on adults, let's find that again!! Oh, and watch this video!


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The Free Library > Health > Health, general > Journal of Immunology Research > January 1, 2017
The Free Library > Date > 2017 > January > 1 > Journal of Immunology Research
Adoptive Cell Therapy of Induced Regulatory T Cells Expanded by Tolerogenic Dendritic Cells on Murine Autoimmune Arthritis.
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Rheumatoid arthritis (RA) is an autoimmune disease causing chronic inflammation of the synovial joints. The inflammatory processes occurring in RA result in hyperplasia of the synovial membrane and infiltration of monocytes, macrophages, T and B cells, mast cells, and dendritic cells (DCs) [1]. Pharmacological therapies for RA include analgesics and anti-inflammatory steroids, which halt the progression of RA but do not cure it. Currently, a curative treatment has yet to be found. Therefore, the development of novel antirheumatic therapies that specifically target aberrant immune processes, dampen inflammation, and promote tolerance is needed.
Recently, cellular therapy for autoimmune diseases has attracted much attention, and as the master regulators of all immune responses, regulatory T cells (Tregs) are the most promising candidates for cell therapy. Natural Tregs (nTregs) are primarily derived from the thymus, and induced Tregs (iTregs) are differentiated from [CD4.sup.+] [CD25.sup.-] [Foxp3.sup.-] T cells in the periphery or in vitro, both of which maintain immunological tolerance and may prevent a variety of autoimmune rheumatic diseases [2, 3]. According to previous reports, iTregs induced by TGF-[beta] in vitro, but not nTregs, retain Foxp3 expression and immunosuppressive activity in the inflammatory microenvironment [4]. In addition, iTregs have been shown to suppress bone erosion and other clinical measures of disease progression in the well-established collagen-induced arthritis (CIA) mouse model of human RA [5, 6], suggesting that iTregs may be therapeutically beneficial for RA [7].
However, culturing iTregs for a period of 5 days has been reported to result in high levels of cell death (detected using propidium iodide staining) [8]. As shown in the study by Kong et al., 3 x [10.sup.6] iTregs per mouse (20 [+ or -] 2g/mouse) were required to significantly inhibit established CIA [9]. The numbers of iTregs induced by TGF-[beta] alone during conventional iTreg culture are not sufficient to satisfy therapeutic demands. Furthermore, after induction by TGF-[beta], only approximately 60% of [CD4.sup.+] T cells express Foxp3 [8, 9], a percentage that is too low to meet the clinical requirements for relevant cell purity, although the remaining [CD4.sup.+] [Foxp3.sup.-]cells did not appear to have any pathogenic effects [10].
Fortunately, tolerogenic dendritic cells (tDCs), which typically present low levels of self-peptide-MHC complexes (signal 1) coupled with limited cell surface expression of costimulatory molecules (signal 2) and secretion of proinflammatory cytokines (signal 3), have been reported to potentially induce tolerance [11]. As the most potent antigen-presenting cells (APCs), DCs are regarded as key instigators or regulators of innate and adaptive immunity. Conventional DCs have the unique ability to activate or promote immune responses depending on their maturation status, whereas tDCs have the ability to induce activated T cell energy and apoptosis and generate/expand nTregs or iTregs in vivo or in vitro. Thus, tDCs are likely to act as both stimulators and inducers to further increase cell numbers and enhance Foxp3 expression in a mixed iTreg population [12].
In this study, we established a new polyclonal expansion method for the generation of iTregs. This method represents the first strategy for generating tDCs induced by IL- 10/TGF/3 in the Treg-induction/expansion system. Mature tDCs (mtDCs), which retained the tolerogenic functions of tDCs and had a stronger expansive ability than tDCs, were employed as the stimulator/inducer. We used mtDCs to successfully expand iTregs, while retaining their regulatory phenotype and potent suppressor functions. These mtDC-expanded iTregs ([iTreg.sub.mtDC]) were associated with a significant reduction in cytokine and CII-directed antibody secretion, polarization of the Treg/Th17 balance, and more effective inhibition of CIA than iTregs. Our findings suggest the potential use of [iTreg.sub.mtDC] as a therapy for autoimmune arthritis.
2.1. Mice. Wild-type male DBA/1J (D1) mice (8 weeks old) were obtained from the Shanghai Laboratory Animal Center of the Chinese Academy of Science (SLACCAS, China). All mice were housed in a pathogen-free environment.
2.2. Ethics Statement. This study was conducted in strict accordance with the recommendations in the guidelines of the Institutional Animal Care and Use Committee of the Chinese Association for Laboratory Animal Sciences. The protocol was approved by the Committee on the Ethics of Animal Experiments of the Shanghai Blood Center (permit number: SBC-IRB-2013-07). All surgery was performed under diethyl ether anesthesia, and all efforts were made to minimize suffering.
2.3. Induction and Evaluation of CIA. CIA was induced in D1 mice via a subcutaneous injection of bovine type II collagen (CII, Chondrex, Redmond, WA, USA) emulsified with an equal volume of complete Freund's adjuvant (Difco, Detroit, MI, USA) on day 0. On day 21, mice received the next injection of 50 [micro]g of CII in incomplete Freund's adjuvant (Difco). The onset of CIA was confirmed on day 25. From day 21 to day 49 after the first immunization, mice were scored every two days for clinical evidence of arthritis of the limb joints by a macroscopic examination. Limb joint arthritis was scored using an established scoring system [13] as follows: no detectable arthritis, 0; erythema and mild swelling, 1; mild erythema and mild swelling involving the entire paw, 2; severe swelling and redness from the ankle to digits, 3; and maximal swelling and redness or obvious joint destruction associated with visible joint deformity or ankylosis, 4. The clinical scores for each mouse are presented as the sum of the scores for the four limbs, and the maximum score for each mouse was sixteen. Two independent observers without knowledge of the experimental protocol performed the scoring. During the 49-day observation period, the arthritis scores increased quickly and constantly and CIA developed in up to 90% of mice.
2.4. Preparation of DCs. DCs were prepared as described [13]. In brief, D1-derived bone marrow (BM) cells were collected and, after red cell lysis, seeded at a density of 5 x [10.sup.5] cells/ml in RPMI 1640 medium supplemented with 10% fetal bovine serum (Invitrogen). On day 0, murine GM-CSF (20ng/ml, PeproTech) was added to the cultures, following which the cultures were pulsed with fresh medium, GMCSF, murine IL-10 (15 ng/ml, PeproTech), and human TGF-[beta]1 (15ng/ml, PeproTech) on days 4 and 7. After 10 days in culture, nonadherent cells were collected and [CD11c.sup.+] cells were sorted as the tDCs. mtDCs were harvested after stimulation with 100 ng/ml lipopolysaccharide (LPS, Sigma-Aldrich) during the final 48 h of culture. Immature DCs (iDCs) were propagated in the presence of GM-CSF alone under the same conditions, and mature DCs (mDCs) were generated following exposure to LPS for 48 h.
All types of DCs were stained with anti-mouse CD11c-APC, CD86-FITC, CD80-PE, and IA-IE-PE mAbs (BD Biosciences Pharmingen) for phenotypic analysis by flow cytometry. Cytokine expression was determined using real-time quantitative PCR. Primer sequences (in the 5; to 3; orientation) were IL-12 p40, GGAAGCACGGCAGCAGAA TA and AACTTGAGGGAGAAGTAGGAATGG; TGF-[beta], TTGCTTCAGCTCCACAGAGA and TGGTTGTAGAGGG CAAGGAC; IL-10, CCAAGCCTTATCGGAAATGA and T TTTCACAGGGGAGAAATCG; and [beta]-actin, ATCCGTAA AGACCTCTATGC and ACACAGAGTACTTGCGCTCA. PCR results were normalized to the expression of the housekeeping gene [beta]-actin [14]. Data shown are a representative of six independent experiments.
2.5. Ex Vivo Generation of [iTreg.sub.mtDC]. iTregs were derived from [CD4.sup.+] [CD25.sup.-] T cells that were purified from the splenocytes of D1 mice using a [CD4.sup.+] [CD25.sup.+] regulatory T cell isolation kit (Miltenyi Biotec, Germany) and stimulated with an anti-CD3/CD28 mAb (50ng/ml, PeproTech) in the presence of TGF-[beta]1 (5ng/ml, PeproTech) and IL-2 (100ng/ml, PeproTech) for 5 days [8]. Then, [iTreg.sub.mtDC] or [iTreg.sub.mDC] were generated by expanding iTregs for 4 days using mtDCs or mDCs at a 5 : 1 ratio (5 T cells : 1 DC). The different types of iTregs were collected, counted, incubated with anti-mouse CD4-FITC and CD25-PE-Cy5 mAbs, fixed, permeabilized, and stained with antimouse [Foxp3.sup.-]PE or isotype control mAbs (BD Biosciences Pharmingen). Then, the coexpression of CD25 and Foxp3 was determined by flow cytometry.
2.6. Proliferation Assay. In the classical polyclonal proliferation assay, responder cells were [CD4.sup.+] T cells, which were isolated from splenocytes of D1 mice using a [CD4.sup.+] T Cell Isolation Kit (Miltenyi Biotec, Germany). The [CD4.sup.+] T cells were labeled with carboxyfluorescein succinimidyl ester (CFSE; Invitrogen, Germany) and stimulated with antiCD3/28 mAbs (50ng/ml, PeproTech). Then, different ratios of iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] (as suppressor cells) were added to the responder cultures. Four days later, the cells were harvested and analyzed by flow cytometry.
In addition, [CD4.sup.+] T cells were isolated from splenocytes of mice with CIA (on day 25 after the primary immunization) and used in the antigen-specific [CD4.sup.+] T cell proliferation assay. Mature DCs were induced as described above and loaded with CII, as stimulator APCs. CFSE-labeled responding [CD4.sup.+] T cells were activated following coculture with CII-loaded mDCs. Then, different ratios of iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] (as suppressor cells) were added to the responder cultures. Four days later, the cells were harvested and analyzed using flow cytometry.
2.7. Evaluation of CIA after [iTreg.sub.mtDC] Treatment. At the onset of CIA, 1 x [10.sup.6] iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] were adoptively transferred into CIA-recipient mice (n = 10 per group) via the tail vein. Control mice were treated with PBS alone. Mice were scored using an established scoring system from days 21 to 49 after the primary immunization [13].
2.8. Histology. The hind paws of iTreg-treated, [iTreg.sub.mtDC]-treated, and CIA mice were collected on day 49 after the primary immunization, and the tissues were stained with hematoxylin and eosin (H&E) and Safranin O. Two independent observers who were blinded to the experimental groups examined the paw sections using a four-point scale: normal, 0; inflammatory infiltrates and synovial hyperplasia, 1; pan nus formation and cartilage erosion, 2; and import cartilage erosion and bone destruction, 3. This global histological score reflected both synovitis (synovial proliferation and inflammatory cell infiltration) and joint destruction (bone and cartilage thickness, irregularity, and the presence of erosions) [13].
2.9. Analysis of Serum Anti-CII Antibody and Cytokine Levels Using CBA and ELISA. Sera were obtained from iTreg-treated, [iTreg.sub.mtDC]-treated, and CIA mice 49 days after the primary immunization and stored at -80[degrees]C. The serum levels of IFN-[gamma], TGF-[alpha], IL-17, IL-6, and IL-10 were determined using a mouse CBA Kit (BD Biosciences Pharmingen) and analyzed by flow cytometry. The concentration of TGF-[beta] was assessed using the mouse TGF-[beta] Platinum ELISA kit (eBioscience), and the concentrations of anti-CII antibodies were measured using a standard sandwich ELISA (Chondrex, Redmond, WA, USA) according to the manufacturer's instructions. Five samples from each group were analyzed.
2.10. Analysis of Treg/Th17 Subsets in Mice with CIA. T cells were isolated from the spleen and inguinal lymph nodes of the iTreg-treated, [iTreg.sub.mtDC]-treated, and CIA mice on day 49 after the primary immunization. These cells were incubated with an anti-mouse CD4-FITC mAb, fixed, permeabilized, stained with anti-mouse FoxP3-PE, IL-17-PE, or isotype control mAbs (BD Biosciences Pharmingen), and then assessed by flow cytometry.
2.11. Th17 Cell Differentiation. Naive [CD4.sup.+] T cells were sorted from the splenic cells of mice with CIA, stained with CFSE as described above, and stimulated with an anti-CD3/CD28 mAb (50ng/ml, PeproTech), IL-1[beta] (10ng/ml, PeproTech), IL-6 (25ng/ml, PeproTech), and TGF-[beta]1 (1ng/ml, PeproTech). Then, iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] were added to the culture at a 1: 1 ratio. After 4 days of coculture, the cells were harvested, incubated with an anti-mouse CD4-APC mAb, fixed, permeabilized, and stained with an anti-IL-17A-PE mAb (BD Biosciences Pharmingen). The production of IL-17A in the cell culture supernatant was measured using the mouse cytokine cytometric bead array (CBA, BD Biosciences Pharmingen) according to the manufacturer's instructions.
2.12. Statistical Analyses. Results were analyzed using GraphPad Prism 5.0 software (GraphPad Software, San Diego, CA, USA), and data are expressed as means [+ or -] standard errors of the means (SEM). Student's t-test was used to assess statistically significant differences between two paired groups, and an alpha value of P < 0.05 was considered statistically significant.
3.1. BM-Derived DCs Induced by IL-10 and TGF-[beta]1 Maintain the Tolerogenic Surface Phenotype and Express Substantial Levels of "Immunosuppressive" Cytokines. According to previous reports, tDCs derived from mouse BM are induced to differentiate with GM-CSF, IL-10, and TGF-[beta]1, which does not affect DC development from replicating BM progenitors [15]. In the present study, a large proportion of induced BM cells were positive for CD11c (>80%), indicating that these cells exhibited a DC phenotype. MHC molecules (IA-IE) and costimulatory molecules (CD86 and CD80) were expressed at low levels on tDCs. Following stimulation with LPS for 48 h, the mean fluorescence intensity (MFI) of each of these molecules was consistently lower on the mature tDCs (mtDCs) than on the mature DCs (mDCs) (Figure 1(a)). Thus, tDCs were comparatively resistant to maturation in response to LPS stimulation and retained the tolerogenic surface phenotype. Additionally, cytokine production by DCs was assessed using real-time PCR. The tDCs displayed higher levels of "immunosuppressive" cytokines, such as IL-10 and TGF-[beta], than iDCs. These cytokines were still expressed at high levels after tDCs were stimulated with LPS, although the expression of the TGF-[beta] mRNA in mtDCs was lower than, but not significantly different from, that in tDCs. However, IL-12 p40 production by tDCs was negligible, even upon stimulation with LPS (Figure 1(b)). There was no significant difference on induction efficiency and cell yields between iDCs, mDCs, tDCs, and mtDCs, though the frequency of [CD11c.sup.+] cells were increased after LPS stimulation.
Based on these data, tDCs induced by GM-CSF, IL-10, and TGF-[beta]1 maintained a tolerogenic surface phenotype, even after LPS stimulation, and expressed substantial levels of "immunosuppressive" cytokines.
3.2. iTregs Generated after mtDC-Mediated Expansion Retain Foxp3 Expression and Potently Suppress Polyclonal [CD4.sup.+] T Cell Proliferation In Vitro. Our iTreg culture strategy consisted of two expansion cycles: (1) the induction of iTregs and (2) the expansion of iTregs by mtDCs (Figure 2(a)). First, [CD4.sup.+] CD25- T cells that were purified from splenocytes of D1 mice were stimulated with an anti-CD3/CD28 mAb (50ng/ml, PeproTech) in the presence of TGF-[beta]1. After 5 days, these cells, "iTregs," were harvested. Then, iTregs were cocultured with mtDCs at a 5: 1 ratio. After 4 days, the cells, termed "[iTreg.sub.mtDC]," were collected, and a significant increase in the total cell number was observed. According to the FACS results, the intracellular expression of Foxp3 in [iTreg.sub.mtDC] was markedly higher (82.0 [+ or -] 3.6%) than that in iTregs (54.9 [+ or -] 2.5%). Using our new culture method, we obtained 3.52-4.3 x [10.sup.6] [Foxp3.sup.+] cells from the 2 x [10.sup.6] [CD4.sup.+] [CD25.sup.-] T cells that were originally plated and included approximately 0.02 x 106 [Foxp3.sup.+] cells (Figure 2(b)). In other words, the [Foxp3.sup.+] cells had expanded approximately 150-fold compared with original cells. However, the [iTreg.sub.mDC], which were expanded by mDCs, appeared to expand more rapidly but generated a lower percentage of cells that expressed Foxp3 (37.4 [+ or -] 4.1%) than the [iTreg.sub.mtDC].
iTregs exhibit immunosuppressive activity. Therefore, the classical polyclonal effector T cell proliferation assay was used as a functional readout to show the inhibitory capacity of iTregs expanded by mtDCs. In this assay, [CD4.sup.+] T cells (responders) were isolated from splenocytes of D1 mice and stimulated with anti-CD3/28 mAbs. As shown in Figure 2(c), the responder cells underwent vigorous proliferation in the absence of suppressors, generating large numbers of dividing T cells in culture. However, this vigorous proliferation was suppressed by the presence of iTregs, [iTreg.sub.mtDC], or [iTreg.sub.mDC] (suppressors). Significantly, [iTreg.sub.mtDC] inhibited [CD4.sup.+] T cell proliferation more effectively than iTregs or [iTreg.sub.mDC] at all the tested suppress-or-to-responder (S:R) cell ratios. Even the lowest dose of [iTreg.sub.mtDC] (an S: R ratio of 1: 8) resulted in the effective inhibition of [CD4.sup.+] T cell expansion, whereas both iTregs and [iTreg.sub.mDC] showed weaker suppression at the same S : R ratio.
Based on these results, iTregs were expanded by mtDCs and maintained the stronger regulatory phenotype and more effective inhibitory potency.
3.3. In CIA Mice, [iTreg.sub.mtDC] Exhibited More Potent Antiarthritic Activity than iTregs. As shown in previous studies, iTregs exert protective effects on established arthritis [4, 9]; therefore, we determined the capacity of [iTreg.sub.mtDC] to improve established CIA. Mice with CIA were injected with the same dose (1 x [10.sup.6] cells/animal) of either iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC], and then, the arthritic index and histopathology were examined to assess the antiarthritic activities of these three types of iTregs.
According to the arthritic scores, all types of iTregs markedly decreased the incidence of CIA and reduced the severity of arthritis compared with that of the untreated control group. More importantly, the injection of [iTreg.sub.mtDC] almost completely inhibited the progression of arthritis during the first 10 days, and subsequently, the arthritic severity never reached the level observed in the CIA control group. When compared with the therapeutic effects of iTregs and [iTreg.sub.mDC] at any time point, treatment with the same doses of [iTreg.sub.mtDC] inhibited the development of CIA to a greater extent, which was substantiated by both the clinical scores and the incidence of arthritis (Figures 3(a) and 3(b)). During the 4-week observation period, [iTreg.sub.mtDC]-treated mice had the mildest arthritic symptoms and the lowest arthritis scores compared with those mice treated with iTregs or [iTreg.sub.mDC].
Next, histopathological specimens of the joints of mice with CIA treated with [iTreg.sub.mtDC] showed the least significant cartilage destruction and least inflammatory cell infiltration compared with those of control CIA mice, although the iTreg or [iTreg.sub.mDC] treatment also reduced the joint damage and inflammation to some extent (Figure 3(c)). The results of the histological examinations were consistent with the resulting clinical scores (Figure 3(d)).
Thus, these in vivo observations confirmed that treatment of CIA mice with [iTreg.sub.mtDC] inhibits the progression of CIA more effectively than the iTreg or [iTreg.sub.mDC] treatment.
3.4. [iTreg.sub.mtDC]-Mediated Inhibition of CIA Was Associated with Modulated Cytokine and Anti-CII Antibody Secretion and the Polarization of the Treg/Th17 Balance. The amelioration of CIA in mice following the iTreg treatment has been attributed to the immune-modulating properties of iTregs [4]. Therefore, we sought to characterize the mechanism by which [iTreg.sub.mtDC] modulates the disease.
First, the secretion of IFN-[gamma], TGF-[alpha], IL-17, IL-6, IL-10, and TGF-[beta] in the sera of CIA mice treated with various iTregs was analyzed on day 49 after the first immunization. The production of IFN-[gamma] and IL-10 significantly increased in [iTreg.sub.mtDC]-treated mice compared with those productions in iTreg- or [iTreg.sub.mDC]-treated mice. In addition, the levels of TNF, IL-17, and IL-6 were significantly reduced and TGF-[beta] was markedly elevated in iTreg-treated, [iTreg.sub.mDC]-treated, or [iTreg.sub.mtDC]-treated mice compared with CIA mice, though there were no significant differences on the levels of those cytokines between the groups treated with different iTregs (Figure 4(a)). Clearly, the attenuation of CIA in mice following adoptive transfer of iTregs was a consequence of their immune modulatory effect on the secretion of various cytokines, and [iTreg.sub.mtDC] showed the strongest modulating effect.
Additionally, the importance of antibodies in the development of CIA pathology is also well described [16]. The serum levels of total anti-CII-specific immunoglobulin (anti-CII IgG) in [iTreg.sub.mtDC]-treated mice were significantly lower than the levels observed in mice with CIA on day 49 following the first immunization, whereas treatment with either iTregs or [iTreg.sub.mDC] failed to obviously reduce anti-CII IgG levels. And there was no significant difference on the levels of anti-CII IgG between [iTreg.sub.mtDC]-treated mice and iTregs- or [iTreg.sub.mDC]-treated mice (Figure 4(b)). Thus, [iTreg.sub.mtDC] inhibited anti-CII-specific antibody responses more powerfully than iTregs or [iTreg.sub.mDC], which contributed to the suppression of the development of CIA.
Furthermore, the Treg/Th17 ratio is considered an important indicator of the severity of CIA. Thus, we determined the percentages of Th17 ([CD4.sup.+] [IL-17.sup.+]) and Treg ([CD4.sup.+] [Foxp3.sup.+]) cells among [CD4.sup.+] T cells isolated from the spleen or inguinal lymph nodes of CIA mice that were treated with various iTregs on day 49 after the first immunization. As shown in Figure 4(c), the Foxp3: IL-17 ratio increased due to an increase in the frequency of Treg cells and a decrease in the frequency of Th17 cells following all types of iTreg treatment. Obviously, the highest Foxp3: IL-17 ratio was detected in [iTreg.sub.mtDC]-treated mice compared with that in either iTreg- or [iTreg.sub.mDC]-treated mice. Thus, [iTreg.sub.mtDC] exerted the most significant modulatory effects on the polarization of the Treg/Th17 balance in vivo.
Taken together, compared to iTregs and [iTreg.sub.mDC], the adoptive transfer of [iTreg.sub.mtDC] exerted the most obvious suppression of cytokine secretion, reduction of CII-directed antibodies, and the most significant polarization of the Treg/Th17 balance.
3.5. [iTreg.sub.mtDC] Inhibit the Antigen-Specific Proliferation of CIA-[CD4.sup.+] T Cells and Suppress Th17 Cell Differentiation In Vitro More Effectively than iTregs. We utilized an in vitro antigen-specific proliferation assay to simulate the in vivo response as a functional readout and to demonstrate the ability of [iTreg.sub.mtDC] to suppress the antigen-specific proliferation of CIA-[CD4.sup.+] T cells. In this assay, [CD4.sup.+] T cells (responders) were isolated from splenocytes of CIA mice and stimulated with CII-loaded mDCs (stimulators). As shown in Figure 5(a), a large number of CIA[CD4.sup.+] T cells underwent antigen-specific amplification after expansion by CII-loaded mDCs. However, no matter if iTregs, [iTreg.sub.mtDC], or [iTreg.sub.mDC] (suppressors) were added to the proliferation assay, this strong proliferation was obviously inhibited. At all tested S : R ratios, [iTreg.sub.mtDC] inhibited antigen-specific [CD4.sup.+] T cell expansion more effectively than iTregs. Notably, a significant difference between the abilities of [iTreg.sub.mtDC] and [iTreg.sub.mDC] to inhibit the antigen-specific [CD4.sup.+] T cell expansion was only observed when they were added at a low dose (at an S: R ratio of 1: 8); iTregs showed weaker suppression at the same S : R ratio.
Given the close relationship between Tregs and Th17 cells [17] and our observations that the adoptive transfer of iTregs decreased the number of Th17 cells and subsequent polarization of the Treg/Th17 balance in the CIA mice, we next investigated whether [iTreg.sub.mtDC] could maintain suppression of Th17 differentiation in vitro, which simulated the in vivo response. In the Th17 induction system, Th17 cells were differentiated from CIA-[CD4.sup.+] T cells by stimulation with IL-6, IL-1[beta], and TGF-[beta] for 4 days. The 1:1 ratio of [iTreg.sub.mtDC] to CIA-[CD4.sup.+] T cells markedly reduced the fraction of cells that expressed intracellular IL-17A from 28.12 [+ or -] 4.79% to 4.83 [+ or -] 1.53%. The addition of the same ratio of iTregs and [iTreg.sub.mDC] to [CD4.sup.+] T cells achieved a weaker suppression of intracellular IL-17A expression (Figure 5(b)). Meanwhile, the level of soluble IL-17A, the key cytokine which Th17 secreted, detected in the supernatant also decreased significantly upon the addition of [iTreg.sub.mtDC] (Figure 5(c)).
Based on these results, [iTreg.sub.mtDC] were the most effective at inhibiting antigen-specific CIA-[CD4.sup.+] T cell proliferation, suppressed Th17 cell differentiation to a greater extent and decreased the secretion of IL-17 in vitro compared with iTregs or [iTreg.sub.mDC], providing strong evidence that adoptive transfer of [iTreg.sub.mtDC] reflected more potent antiarthritic activity in the CIA mice.
[CD4.sup.+] CD25+ [Foxp3.sup.+] regulatory T cells play a crucial role in maintaining immune tolerance. As shown in multiple studies, nTregs prevent the appearance and development of autoimmune diseases in many animal models [18, 19]. However, nTregs are unable to treat established autoimmune diseases because of their instability in an inflammatory milieu [20-22]. In addition, the widespread use of nTreg-based therapy is hindered by the low frequency and lack of reliable surface markers for purification [23, 24]. In 2002, Zheng et al. were the first to report that TGF-[beta] induces the differentiation of [CD4.sup.+] CD25- cells into [CD4.sup.+] [CD25.sup.+] Tregs (iTregs) in vitro [25]. Over the past decade, a number of studies on iTregs and their relationship to the regulation of immunity and the outcomes of autoimmune diseases have shown that adoptive transfer of [Foxp3.sup.+] iTregs ameliorates autoimmune disease in mouse models of lupus, gastritis, diabetes, and RA [4, 26-28], although the capacity to apply this therapy in the clinic remains controversial [29, 30]. In this study, we established a new method for the polyclonal expansion of iTregs using mtDCs and assessed the suppressive capacity of [iTreg.sub.mtDC] in vitro and in CIA mice. According to our results, [iTreg.sub.mtDC] corrected defects in the number and function of iTregs in vivo and in vitro and thus possessed the potential to be a powerful tool in the treatment of autoimmune diseases.
In the classical iTreg culture method, iTreg differentiation is induced by an anti-CD3/CD28 antibody and TGF-[beta]. This method produces few cells due to a high cell death rate, and the number of cells obtained is not sufficient to meet therapeutic demands. We modified the iTreg-induction/culture protocol to produce more cells and to promote the clinical application of iTreg-based therapy. In the conventional polyclonal expansion method, an anti-CD3/28 antibody or mature DCs are used as stimulators, and the addition of high doses of anti-CD3/28 antibody led to severe cell apoptosis caused by excessive activation, as the anti-CD3/28 antibody preferentially amplified [CD4.sup.+] effector cells and not iTregs. Mature DCs had a similar effect. Fortunately, semimature DCs [31] or DCs with a tolerogenic phenotype [32], such as the low expression of MHC or/and costimulatory molecules, had the ability to expand or generate Tregs de novo and promote the apoptotic death of effector T cells in some studies. Tolerogenic DCs were the optimal candidate to amplify the [Foxp3.sup.+] iTregs, particularly in a mixed population of cells.
DCs have the unique ability to activate or suppress immune responses depending on their maturation status, phenotype, and tissue of origin. Due to their inherent plasticity, DCs are regarded as key instigators or regulators of innate and adaptive immunity. It is commonly held that conventional DCs have the unique ability to activate or suppress immune responses depending on their maturation status, whereas tolerogenic DCs exhibit the distinct functional properties in promoting tolerance. tDCs describe a broad range of immunoregulatory DCs that are usually immature [33], exhibit a plasmacytoid morphology [34], or are alternatively activated [15]. And on the basis of traditional definition, Jiang et al. [35] further hierarchically arranged two classes of maturation programs for DCs. They found that DCs matured by E-cadherin contained the phenotypic hallmarks of conventional mature DCs but without the secretion of inflammatory cytokines. And these E-cadherin-matured DCs could induce peripheral T cell tolerance in vivo and protect against EAE. Thus, alterations in adhesion could be seen as producing "phenotypically mature" immunogenic DCs, whereas microbial stimuli yield "functionally mature" immunogenic DCs under conditions of inflammation. Also, Vander Lugt et al. [36] provided a gene regulatory framework and indicated that the presence of DCs in peripheral tissues appeared phenotypically mature and had a critical role in peripheral tolerance, which illuminated the molecular underpinnings of DC maturation and function. Meanwhile, Guilliams et al. [37] provided a universal toolbox for the automated identification of DCs using unsupervised analysis of flow cytometry, paving the way toward the faithful identification of DC subsets to grasp the fascinating functional heterogeneity of DCs.
In this study, DCs exposed to IL-10 and TGF-[beta] were resistant to maturation in response to LPS stimulation. The MFI of IA-IE, CD80, and CD86 expression on mtDCs was lower than the MFI on mDCs. Moreover, mtDCs secreted higher levels of IL-10 and TGF-[beta] than mDCs, whereas IL12 p40 production by mtDCs was negligible. Considering the limited cell expansion capability of tDCs, mtDCs matured by LPS were employed in this new method. mtDCs also displayed the tolerogenic surface phenotype and secreted a large number of "immunosuppressive" cytokines, whose tolerogenic characteristics were not significantly different from those of tDCs. More importantly, mtDCs effectively expand T cells, but tDCs always show a poor capacity to activate T cells. Due to their stable tolerogenic functions and improved ability to expand iTregs, mtDCs were employed as the best stimulator/inducer in this study. As a positive control, conventional mDCs were also added to expand iTregs in parallel experiments. As expected, fewer cells expanded by mDCs ([iTreg.sub.mDC]) expressed Foxp3 (37.4 [+ or -] 4.1%) than cells before amplification. Interestingly, these [iTreg.sub.mDC] suppressed effector [CD4.sup.+] T proliferation to a greater extent than iTregs in vitro, possibly because iTregs have enhanced functional activities after activation. Similarly, activated nTregs exert a stronger suppressive effect than silent nTregs [20]. This reason may be why [iTreg.sub.mtDC], which are activated further by mtDCs, exerted more potent inhibitory effects than iTregs induced by TGF-[beta] alone.
Noticeably, although more than 80% of the cells were [Foxp3.sup.+] after the combined induction/expansion with TGF-and mtDCs, approximately 10% [CD4.sup.+] [Foxp3.sup.-] cells and less than 5% [CD11c.sup.+] mtDCs were observed. So [iTreg.sub.mtDC] were the mixture of [CD4.sup.+] [Foxp3.sup.+] cells, [CD4.sup.+] [Foxp3.sup.-] cells, and mtDCs. Lan et al. compared the suppressive effects of a mixed population of iTregs and [Foxp3.sup.+] iTregs and showed that these activated [CD4.sup.+] [Foxp3.sup.-] cells did not develop the ability to exert pathogenic effects after TGF-[beta] treatment [10]. Thus, the purified [Foxp3.sup.+] Treg population may not need to be sorted. Meanwhile, established murine arthritis is significantly inhibited by a tDC infusion [38, 39]. As shown in our previous study, mtDCs exerted suppressive effects in vitro and on the CIA mouse model after IL-10 and TGF-fi polarization [13]. In this study, we did not isolate [iTreg.sub.mtDC] from mtDCs; therefore, we cannot exclude the possibility that the very limited number of mtDCs infused with [iTreg.sub.mtDC] may have contributed to the suppression of CIA.
Multiple mechanisms are involved in the inhibitory activities of iTregs in vivo. Our study has confirmed that different types of iTregs suppressed CIA in mice through similar mechanisms, including modulation of cytokine secretion, prolonged inhibition of anti-CII IgG antibodies, and polarization of the Treg/Th17 balance. Following treatment with [iTreg.sub.mtDC], the serum levels of TGF-[alpha], IL-17, and IL-6 were significantly reduced, whereas the levels of IFN-[gamma], IL-10, and TGF-[beta] were markedly elevated in mice with CIA compared with the levels in mice treated with iTreg or [iTreg.sub.mDC]. In addition, [iTreg.sub.mtDC] inhibit anti-CII-specific antibody responses more powerfully than iTregs or [iTreg.sub.mDC], which contribute to the suppression of the development of CIA. Interestingly, the levels of IFN-[gamma], which is mainly produced by NK cells and Th1 cells and induces inflammation, were increased after the adoptive transfer of iTregs. In fact, an increasing number of studies have recently confirmed that IFN-[gamma] acts as a disease-limiting factor in CIA and not a pathogenic factor [40, 41], though several studies have recently revealed a role for IFN-a in the pathogenesis of a subset of RA patients [42]. The protective mechanism of IFN-[gamma] is to inhibit the differentiation of monocytes/macrophages into osteoclasts and suppress IL-1fi-mediated MMP1 and MMP3 production by synovial fibroblasts in antigen-induced arthritis, thereby limiting cartilage degradation. More importantly, IFN-[gamma] facilitates the development of Tregs from precursor T cells and increases their suppressive effects on CIA. Conversely, IFN-[gamma] inhibits Th17 cell development and suppresses their effector functions. Therefore, the high level of IFN-[gamma] may reduce inflammation and osteoclast differentiation in CIA mice. Additionally, another therapeutic benefit of iTregs is attributed to the fact that they express low levels of the IL-6 receptor and subsequent STAT-3 phosphorylation [43], express high levels of the Bcl-2 gene, and resist apoptosis [25], and, most importantly, iTregs shift the Treg/Th17 balance to a Treg-predominant phenotype [9]. Thus, a tolerant state was established following the iTreg treatment in vivo, which promoted Treg cell differentiation and proliferation while limiting the differentiation of Th17 cells in vivo. As expected, compared with iTregs or [iTreg.sub.mDC], [iTreg.sub.mtDC] showed more significant polarization of the Treg/Th17 balance in the CIA mice. Based on our in vitro experiments, [iTreg.sub.mtDC] were the most effective cell type at reducing IL-17A secretion from [CD4.sup.+] T cells and suppressing Th17 cell differentiation. Meanwhile, [iTreg.sub.mtDC] more effectively inhibited antigen-specific [CD4.sup.+] T cell expansion than iTregs at all tested S: R ratios, whereas a significant difference in the inhibition of antigen-specific [CD4.sup.+] T cell expansion between [iTreg.sub.mtDC] and [iTreg.sub.mDC] was only observed at a low S: R ratio (1 : 8). Thus, [iTreg.sub.mtDC] promoted a more potent tolerogenic microenvironment, a feature that was evidenced by the effectively reduced presentation of CIA symptoms and inhibition of CIA progression. Because [iTreg.sub.mtDC] had the strongest antiarthritic effects, we were able to adoptively transfer low doses of [iTreg.sub.mtDC] (1 x [10.sup.6] cells/CIA mouse) compared with the higher doses of iTregs ([greater than or equal to] 3 x [10.sup.6] cells/CIA mouse) required to achieve the same inhibition of established arthritis. Therefore, under the same culture condition, [iTreg.sub.mtDC] meet the requirements for clinical application more readily than other iTregs, in terms of both the [Foxp3.sup.+] cell purity and the number of cells produced.
In general, antigen-specific iTregs are believed to have a stronger suppressive ability than nonspecific iTregs [9]. However, in autoimmune diseases, such as RA and lupus, the specific antigens responsible for the disease processes remain undefined. Polyclonal [iTreg.sub.mtDC] suppressed T cell proliferation and CIA, suggesting that manipulation of polyclonal [iTreg.sub.mtDC] may represent a therapeutic tool with which to combat autoimmune diseases in which specific antigens have not been defined. Moreover, once the specific pathogenic antigens of RA are identified, mtDCs, potent APCs, could acquire and present specific antigens to iTregs, subsequently activating and propagating the antigen-specific iTregs. Thus, we believe that the involvement of mtDCs in the iTreg culture system will greatly improve future iTreg-induction/expansion methods.
In summary, we established a new iTreg polyclonal expansion method. Using this method, iTregs were efficiently expanded by tolerogenic DCs ex vivo to reach clinically relevant cell numbers while retaining and even strengthening their regulatory phenotypes and potent suppressor functions. In our study, these [iTreg.sub.mtDC] prevented established CIA by modulating cytokine secretion, anti-CII antibodies, and the polarization of the Treg/Th17 balance. Based on these results, we confidently postulate that human mtDCs are a suitable stimulator to expand human Tregs to achieve clinically relevant cell numbers and enhance their inhibitory activity in vitro and in vivo, although some differences have been observed between human Tregs and mouse Tregs. Moreover, mtDCs are also potent APCs. mtDCs could acquire/present specific antigens to Tregs and then activate/propagate the antigen-specific Tregs, which are thought to have a stronger suppressive ability and better therapeutic effects than nonspecific iTregs. Thus, we have sufficient reasons to believe that the use of mtDCs in the iTreg culture system may contribute to improve future Treg-induction/expansion methods, establish a large-scale in vitro cell culture system for clinical applications, and enhance the therapeutic effects and extensive use of Treg-based cell therapy in the forthcoming decade.
https://doi.org/10.1155/2017/7573154
The authors declare no financial interest or commercial conflict of interest.
Jie Yang and Lidong Liu contributed equally to this work. Acknowledgments
This work was supported by grants from Shanghai Public Health Construction Projects Foundation (15GWZK0501), Shanghai Natural Science Foundation (13ZR1452100, 15ZR1438300), the Foundation of Shanghai Municipal Commission of Health and Family Planning (201440439, 201640096, 20154Y0192, and 20154Y0079), and China National Natural Science Foundation (81302571). The authors also thank the Department of Animal Science in Shanghai Jiao Tong University School of Medicine for laboratory animal husbandry.
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[27] M. C. Fantini, C. Becker, I. Tubbe et al., "Transforming growth factor beta induced FoxP3+ regulatory T cells suppress Th1 mediated experimental colitis," Gut, vol. 55, pp. 671-680, 2006.
[28] E. Godebu, D. Summers-Torres, M. M. Lin, B. J. Baaten, and L. M. Bradley, "Polyclonal adaptive regulatory CD4 cells that can reverse type I diabetes become oligoclonal long-term protective memory cells," Journal of Immunology, vol. 181, pp. 1798-1805, 2008.
[29] J. Andersson, D. Q. Tran, M. Pesu et al., "CD4+ FoxP3+ regulatory T cells confer infectious tolerance in a TGF-betadependent manner," The Journal of Experimental Medicine, vol. 205, pp. 1975-1981, 2008.
[30] S. Floess, J. Freyer, C. Siewert et al., "Epigenetic control of the foxp3 locus in regulatory T cells," PLoS Biology, vol. 5, no. article e38, 2007.
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[32] J. Yang, Y. Yang, H. Fan, and H. Zou, "Tolerogenic splenic IDO (+) dendritic cells from the mice treated with inducedTreg cells suppress collagen-induced arthritis," Journal of Immunology Research, vol. 2014, Article ID 831054, 2014.
[33] B. M. Fu, X. S. He, S. Yu et al., "A tolerogenic semimature dendritic cells induce effector T-cell hyporesponsiveness by activation of antigen-specific CD4+ CD25+ T regulatory cells that promotes skin allograft survival in mice," Cellular Immunology, vol. 261, pp. 69-76, 2010.
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[42] J. Rodriguez-Carrio, P. Lopez, and A. Suarez, "Type I IFNs as biomarkers in rheumatoid arthritis: towards disease profiling and personalized
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[43] S. G. Zheng, J. Wang, and D. A. Horwitz, "Cutting edge: [Foxp3.sup.+]CD4+CD25+ regulatory T cells induced by IL-2 and TGF-beta are resistant to Th17 conversion by IL-6," Journal of Immunology, vol. 180, pp. 7112-7116, 2008.
Jie Yang, (1) Lidong Liu, (1) Yiming Yang, (1) Ning Kong, (2) Xueyu Jiang, (1) Juan Sun, (1) and Rufeng Xie (1)
(1) Blood Engineering Laboratory, Shanghai Blood Center, Shanghai 200051, China
(2) Division of Rheumatology, Huashan Hospital, Fudan University, Shanghai 200040, China
Correspondence should be addressed to Jie Yang; allareblue@gmail.com
Received 15 January 2017; Revised 1 April 2017; Accepted 27 April 2017; Published 18 June 2017
Caption: Figure 2: iTregs expanded in vitro by mtDCs retained Foxp3 expression and efficiently inhibited [CD4.sup.+] T cell proliferation. (a) Schematic overview of the induction/expansion strategy for iTregs. The two expansion cycles were established as described in the Materials and Methods. The differentiation of iTregs was first induced with an anti-CD3/CD28 mAb, TGF-[beta], and IL-2 for 5 days in vitro. Then, [iTreg.sub.mtDC] or [iTreg.sub.mDC] were generated by expanding iTregs for 4 days using mtDCs or mDCs, respectively. The expression of the transcription factor Foxp3 and CD25 in cells was determined in each cycle using flow cytometry. Data are representative of three independent experiments. (b) Expansion of different types of iTregs was determined by counting the cells, and Foxp3 expression levels were determined by flow cytometry. All data are presented as means [+ or -] SEM (n = 5), and *P < 0.05, **P < 0.01, and ***P < 0.001 for the comparisons of the indicated groups using the unpaired t-test. (c) The ability of iTregs to inhibit the proliferation of [CD4.sup.+] T cells was assessed using the proliferation assay. In this assay, [CD4.sup.+] T cells (responders) were isolated from splenocytes of D1 mice, stained with CFSE, and stimulated with an anti-CD3/CD28 mAb and IL-2. Then, different ratios of iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] (suppressor cells) were added to the responder cultures. Four days later, the cells were harvested and analyzed by flow cytometry. Progressive dilution of CFSE in responder cells was used as a readout of proliferation in the presence or absence of different types of iTregs. The results are presented as means [+ or -] SEM (n = 3). *P < 0.05 and **P < 0.01 compared with the indicated groups using the unpaired t-test. Caption: Figure 3: [iTreg.sub.mtDC] exhibited a more potent antiarthritic activity in CIA mice than iTregs. Following the onset of experimentally induced CIA (on day 25 after the primary immunization), recipient mice received adoptively transferred iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] (1 x [10.sup.6] cells/ animal). Mice were scored for clinical signs of arthritis in the limb joints by macroscopic examination three times per week. Limb joint arthritis was assessed using an established scoring system. The mice were sacrificed on the 49th day after the first immunization with CII. (a) Arthritic scores and incidence for each group following the adoptive transfer of different types of iTregs (n = 10) during the observation period are shown. *P <0.05, **P <0.01, and ***P <0.001 for the comparison of the CIA group and the CIA + [iTreg.sub.mtDC] group; #P <0.05 for the comparison of the CIA + iTreg group and the CIA + [iTreg.sub.mtDC] group; and $P <0.05 for the comparison of the CIA + [iTreg.sub.mDC] group and the CIA + [iTreg.sub.mtDC] group using the unpaired t-test. (b) Hind paw specimens were collected from recipient mice treated with different types of iTregs on the 49th day after the first immunization with CII and were stained with H&E (top, synovial joint inflammation) and Safranin O (bottom, cartilage erosion). All specimens were shown at x200. (c) Histopathological scores for inflammation and destruction in each group are expressed as the means [+ or -] SEM of four individual experiments. **P <0.01 for the comparisons of the indicated groups using the unpaired t-test.
Caption: Figure 4: In the CIA mouse model, [iTreg.sub.mtDC] modulated the secretion of cytokines and anti-CII antibodies and polarized the Treg/Th17 balance. Recipient mice that received adoptively transferred iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] were sacrificed on the 49th day after the first immunization with CII, and serum was collected from each group of mice. (a) Serum levels of IFN-[gamma], TGF-[alpha], IL-17, IL-6, and IL-10 were measured using CBA assays, and TGF-[beta] secretion was measured using an ELISA. Data are reported as means [+ or -] SEM (n = 5). *P < 0.05, **P < 0.01, and ***P < 0.001 for the comparisons of the indicated groups using the unpaired t-test. (b) Serum levels of total CII-specific immunoglobulin were determined using an ELISA. The results from five independent replicate experiments were pooled. Data are reported as means [+ or -] SEM (n = 5). *P < 0.05 and **P < 0.01 for the comparisons of the indicated groups using the unpaired t-test. (c) [CD4.sup.+] T cells from the spleen and inguinal lymph nodes of CIA mice treated with or without different types of iTregs were collected, permeabilized, and stained with an anti-Foxp3 mAb and an anti-IL-17 mAb to detect intracellular expression. FACS flow cytometry was used to measure the percentage of positively stained cells, and the ratios of Treg/Th17 cells are expressed as the means [+ or -] SEM of five independent experiments. * P < 0.05 and **P < 0.01 for the comparisons of the indicated groups using the unpaired t-test.
Caption: Figure 5: [iTreg.sub.mtDC] exerted a more potent immunosuppressive effect on antigen-specific [CD4.sup.+] T cell proliferation and suppressed Th17 cell differentiation more effectively. (a) The ability of [iTreg.sub.mtDC] to inhibit the proliferation of antigen-specific [CD4.sup.+] T cells was assessed using the proliferation assay. In this assay, [CD4.sup.+] T cells (responders) were isolated from splenocytes of CIA mice (on day 25 after the primary immunization), stained with CFSE, and stimulated with CII-loaded mDCs (stimulators) derived from the BM of normal D1 mice. Then, different ratios of iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] (suppressor cells) were added to the responder cultures. Four days later, the cells were harvested and analyzed by flow cytometry. The results are shown as means [+ or -] SEM (n = 3). *P < 0.05 and **P < 0.01 for the comparisons of the indicated groups using the unpaired t-test. (b) The suppression of Th17 cell differentiation by coculture with [iTreg.sub.mtDC] was assessed using the standard Th17 cell differentiation system. As described in the Materials and Methods, naive [CD4.sup.+] T cells were isolated from the splenocytes of CIA mice, stained with CFSE, and stimulated with an anti-CD3/CD28 mAb, IL-1[beta], IL-6, and TGF-[beta]1. Then, a 1: 1 ratio of iTregs, [iTreg.sub.mDC], or [iTreg.sub.mtDC] was added to this differentiation system. After 4 days of cocultivation, the percentages of CFSE+ [CD4.sup.+] IL17A+ cells in the different groups were determined by flow cytometry as the percentage of induced Th17 cells. Data are reported as means [+ or -] SEM (n = 3), and *P < 0.05, **P < 0.01, and ***P < 0.001 represent the comparisons of the indicated groups using unpaired t-test. (c) After 4 days of cocultivation, supernatants were collected from the groups mentioned in (b). The concentration of soluble IL-17A in the coculture supernatant was determined using CBA. Data are reported as means [+ or -] SEM (n = 3), and *P < 0.05, **P < 0.01, and ***P < 0.001 represent the comparisons of the indicated groups using the unpaired t-test.
Figure 1: Characteristic profile of tolerogenic DCs derived from D1
mice.iDCs, mDCs, tDCs, and mtDCs were induced to differentiate in
vitro, as described in the Materials and Methods, after which
CD11c+ cells were harvested. (a) These four types of DC were
stained with CD86, CD80, IA-IE (thick lines), or isotype-matched
mAbs (thin lines), and the expression of these markers was analyzed
by FACS using flow cytometry. The frequency of positively stained
cells and MFI of a representative of 10 independent experiments are
shown in the FACS profile. The mean fluorescence intensity is
reported as the mean [+ or -] SEM (n = 10). **P < 0.01 for the
comparison of mDCs with mtDCs using unpaired t-tests. (b) IL-12 p40,
IL-10, and TGF-[beta] expression in DCs was determined by real-time
PCR. All results were normalized to the expression of the housekeeping
gene [beta]-actin and are expressed as the means [+ or -] SEM of six
independent experiments. *P < 0.05 and **P < 0.01 for the
comparisons of the indicated groups using the unpaired t-test.
The mean fluorescence intensity
iDC                      mDC
CD86      336.79[+ or -] 82.55    2474.36[+ or -] 613.39
CD80      340.81[+ or -] 72.05    1445.34[+ or -] 86.32
IA-IE     365.05[+ or -] 96.83    3736.40[+ or -] 910.87
tDC                       mtDC
CD86      246.01[+ or -] 134.23     362.88[+ or -] 86.93 **
CD80      214.98[+ or -] 55.72      277.09[+ or -] 43.73 **
IA-IE     245.79[+ or -] 55.99      336.89[+ or -] 52.27 **
COPYRIGHT 2017 Hindawi Limited
Yang, Jie; Liu, Lidong; Yang, Yiming; Kong, Ning; Jiang, Xueyu; Sun, Juan; Xie, Rufeng
Journal of Immunology Research
Natural Immunomodulators.
CD80 Expressed by [CD8.sup.+] T Cells Contributes to PD-Ll-Induced Apoptosis of Activated [CD8.sup.+] T Cells.
Health aspects
Bone morphogenetic proteins
Transforming growth factors


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text:
Sarah Best
Call Sarah +61 2 8915 1007
Call Sarah +61 488 137 042
"Competition and consumer law can be complex and knowing the ACCC's approach on matters is all important in an ever-changing regulatory landscape. I partner with clients to help them understand these laws so they can make smart, strategic decisions for their businesses."
Sarah has worked in the competition and consumer law space for over 20 years, advising various fast moving consumer goods companies over this time on a wide variety of legal issues. Sarah is a solutions-oriented lawyer who enjoys working with clients to provide them with practical legal advice so they can run their businesses within the limits of the law. She values building connections with people and really understanding her clients' businesses.
Sarah advises on anti-competitive conduct and the impact of competition law on commercial activities like distribution and supply chain arrangements, exclusivity provisions, joint venture/collaboration agreements and intellectual property licensing arrangements.
Sarah also advises on merger clearances and provides related competition law structuring advice, setting up information sharing protocols to manage the flow of information between parties to minimise the risks of gun jumping prior to the close of deals. She has successfully obtained merger clearances for various large domestic transactions across a number of different consumer goods industries.
Sarah advises suppliers including retailers and online platforms on consumer law issues including misleading and deceptive conduct, consumer guarantees, unfair contract terms, advertising and marketing sign-offs and general compliance with the Australian Consumer Law. She also advises on product recalls, both working with clients to help them manage the legal issues that arise in this context and liaising with the responsible regulator.
Sarah represents clients in dealings with the ACCC including investigations and enforcement actions and her strong advocacy and solutions-based approach has led to great outcomes for clients in adversarial and cooperative contexts.
Bega Cheese — obtained merger clearance from the ACCC and provided competition law advice in relation to various transactions including its 2020 acquisition of Lion Dairy & Drinks, owners of the Dairy Farmers, Yoplait, Big M, Masters and Dare and Farmers Union iced coffee brands; its 2018 acquisition of Saputo's Koroit processing facility; and its 2017 acquisition of various assets from Mondelez Australia including the Vegemite and Kraft peanut butter brands. These were significant projects for Bega Cheese because of their size and the complex legal and commercial issues they raised as well as the commercial importance of securing fast and uncontested merger clearance. They also involved the provision of competition law structuring advice and the setting up of information sharing protocols to manage the flow of information between the parties prior to completion.
Bega Cheese Limited/Dairy Inquiry — advised Bega Cheese in responding to a section 95ZK notice issued by the ACCC as part of the Federal Government's enquiry into the dairy industry in Australia. This was a significant project because of the enormous amount of documentation required to be reviewed and produced to respond to the ACCC's notice, requiring the use a mix of human and technological solutions.
The Coca-Cola Company — advised TCCC in relation to the competition law issues associated with a number of transactions including its 2018 acquisition of a 45% interest in the Made Group of Companies (owners of Cocobella, Rokeby Farms and Impressed Juice brand of products) together with its bottler, Coca-Cola Amatil.
Sun Bum — advised Sun Bum in relation to the product recall of one of its sunscreen products in 2022, which involved working with the client's US based recall team and regulatory consultant and liaising with the Therapeutic Goods Administration on all aspects of the recall. This lead to the successful closing-out of the recall within three months.
Decathlon — represented Decathlon in relation to a section 155 notice issued to it by the ACCC, the subsequent ACCC investigation and then proceedings which were commenced by the ACCC against Decathlon arising out of breaches of various mandatory safety standards.
Competition and consumer law review of customer sales agreements of funeral services client — provided competition and consumer law advice on the bundling and pricing provisions in this client's customer sales agreements. This involved a complex assessment of markets for the supply of various goods and services, taking into account the current status quo in 2020 and potential structural reform being slated by the State Government.
Merger and structuring advice for FMCG client — provided competition law advice to a key offshore FMCG client in relation to their majority investment in a local manufacturing company. This involved an in-depth analysis of our client's Australian business and the acquisition target and then setting up protocols and guidelines to be followed locally to ensure compliance with competition law.
Competition law review of commercial supply arrangement for FMCG client — completed an in-depth competition law review for a key client in relation to an exclusivity restraint and its rollover in a core commercial supply agreement with a retailer. This involved an assessment of the purpose and effect of the restraint on competition in various FMCG markets which amongst other things required an analysis of market scan data and consumer behaviour data.
Advice on global buying consortium — advised an FMCG client on a local subsidiary's proposed involvement in a global buying consortium. This involved a complex analysis of various legal and practical issues in order for the competition law risks associated with the proposal to be assessed.
Cartel conduct and resale price maintenance audit for key client — conducted comprehensive and in-depth cartel conduct and resale price maintenance audits for a key client of the firm over multiple years as part of the client's compliance initiatives and in order to assist the client in determining the level of competition law risk exposure within their sales team. These audits involved the review of an enormous number of emails and documentation, requiring the use of a mix of human and technological solutions.
Indigenous Business Australia — advised on a suite of commercial funding agreements with a financial institution and the Federal Government and associated competition law structuring requirements. This involved an analysis of back-to-back contractual arrangements and advising on protocols and guidelines to be followed to ensure compliance with competition law.
Reebok Australia — acted as Compliance Auditor to this client to undertake an annual review of an Australian Consumer Law Compliance Program Reebok Australia as ordered by the Federal Court of Australia arising out of proceedings brought against it by the ACCC for false and misleading advertising.
Colgate-Palmolive and Hill's Pet Nutrition — reviewed various different types of commercial contracts for this client over the years, advising on both general contract law issues as well as consumer law and privacy issues. I also provide strategic competition law advice from time to time in relation to these clients' various supply channels and distribution arrangements and pricing policies.
Member, Competition Law Discussion Group
Member, Law Society of NSW
Member, Law Council of Australia, Business Law Section
Bachelor of Arts/Bachelor of Laws (Honours), The University of Sydney
Addisons' top ten tips for managing a product recall
ACCC slates game-changing package of merger reforms – let the debate begin
Insights: 11 October 2021
My Sarah Best


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Q: .exe generated in cygwin 1.7 is not running on cygwin 1.52 A year back i have developed a small script in perl for a customer and developed its .exe using pp .
we delivered .exe file along with basic cygwin(1.52) install files to run that .exe to customer.
Now we got an enhancement in the script .we lost all dev environment for cygwin .Again we installed fresh cygwin with 1.7 version ,coded and generated .exe file.this file is not running in the customer environment who have cygwin 1.52.Just it will be balnk after executing the .exe
we cannot ask customer to upgarde the test environment.
what is other way to make it run .exe with cygwin dll 1.7 on cygwin 1.52.
Any help would be higly appreciated.
Rgds,
sowm

A: The developers of Cygwin are strict about upward compatibility, but they don't try to provide backward compatibility between major releases, like between 1.5 and 1.7.  This means you can build a program on 1.7 that runs on 1.5 only as long as you avoid calling functions that were added to the Cygwin DLL in 1.7.
Most likely the reason your code calls 1.7-only functions is that it is using libraries that auto-discover platform features.  There could be other reasons, but without any details about what exactly is failing, it's difficult to guess.
If the problem is due to third-party libraries, as I'm guessing, it may be practical to spend time to figure out how to make them revert to the common functionality provided by both 1.5 and 1.7.  For instance, with an autoconf-based system, you can hand-edit the config.h file the configure script produces to turn off use of some discovered features.  This in turn means building all those libraries from source yourself, rather than downloading binary versions from the Cygwin project repository and using them directly.
It may be easier to pull a Cygwin 1.5 environment out of the Cygwin Time Machine.
By the way, you are aware that distributing Cygwin and executables built with it requires that those executables comply with the GPL, or that you buy a Cygwin commercial distribution license, right?  If not see the FAQ.

A: Install 1.5 with http://cygwin.com/setup-legacy.exe

redpajama_set_name: RedPajamaStackExchange


Some teams are better with faster delivery, others with higher quality, and others with higher morale. The list of things that might make at team better is inexhaustible. But what if you wanted it all, not only for your team, but also for your whole organization?
A new book, Company-wide Agility with Beyond Budgeting, Open Space & Sociocracy: Survive & Thrive on Disruption, by Jutta Eckstein and John Buck, looks at how to extend agile beyond the traditional software development group. While every organization could benefit from the ideas presented by Eckstein and Buck, their target audience is those who wish to "achieve revolutionary benefit across the board rather than only in the engineering department."


Listening to what people have to say about movies, restaurants, food, an experience that helps in improving the returns they provide them.
Be it a service improvement or the actual product improvement, feedbacks are valuable to both the customer and the provider. They have the same effect and importance in the employment industry as well.
The importance of the feedback is grouped from who/where it came from and to who it is valuable to. Employee feedback is valuable to the managers and the organization. The intention of feedback is not just gathering data and showing reports that a certain of data has been gathered from the employees over time.
But it is the whole process which helps to understand the employee's pulse, look into the concerns and neglected areas, cue for appreciations, to provide a platform to voice their opinions, and finally, a direction for the management to take steps in and to work on.
Capturing the pulse is not easy with annual feedbacks, the emotion and connectivity may change over time and by the time the feedback is collected the employee may feel no longer the need to voice it out. It is a recurring and timely action that the management needs to look at.
The feedback can help in improving the fundamental areas of the company like the team building, performance, employee-manager dynamics, and employee experience. Let's discuss each of this in a deeper context.
The performance of an employee depends on productivity and satisfaction. There is a direct dependency on the two. When the environment is motivational enough and there is support from the team, the employee will have better satisfaction in terms of his work which in return will promise a better performance rate. Also, pushing the individual to work on a field of disinterest is also a productivity-draining factor.
All this can be solved when the right questions are asked to the employee and they get a platform to voice all of their thoughts.
Say there is a question asking 'What would you like to work on from the next quarter' can provide an opportunity for Ben, a Frontend engineer, to say that he has interest in UI/UX design and that he would love to give it a try.
In the next few days, there can an opportunity for Ben to work on UI/UX as a part-time task. With a few days, the team can again have a discussion on which to pursue based on his best performance. Here, Ben gets to improve his performance and the company earn themselves a great designer. Even if it doesn't work, Ben would have had the opportunity to try his heart making him believe more in the company and go back working with a contented heart. His performance will improve no matter what way as he will strive to do his best.
A Study says most employees leave a job because of their managers or bosses. Generations have tried to make the employee-manager relationship better. The best and easiest way is to show the employees that the managers, founders and the management cares and value the employees. By showing, it doesn't necessarily mean showering them with money but by little actions and steps.
Acknowledgment is a simple yet powerful step. Thanking an employee for doing a good job, motivating an employee when he/she feels low and taking measure to fix the issue the employee has been facing and working with them to make their experience with organization more conducive are ways to communicate that you value them.
Acknowledge that you have received their feedback and then show them that their feedbacks are valuable insights that is making their organization better. When they see that they are being heard, the feedbacks will become more meaningful and real issues are discussed. It is just a matter of practice.
The right use of feedbacks allow for better relationships built on trust and united goals, and the organizations benefit from a value-driven workforce that is more engaged and bring about exemplar results.
For example, Jack has been working on a project for days now and has been handling it single handedly. He feels drained and exhausted and there is still 10 more days to complete the project. But when he walks in to his desk on the next day, he finds a note that says, 'Hey rockstar, good going with the project. Thank you for being awesome'.
Now he works with a smile on his way because he is acknowledged for his effort.
And 4 days later he receives a message saying, 'You need a break. Here is an energy booster' and there is his favorite drink/snack with it. On the day of completion, there is a small gathering where the manager appreciates Jack in front of the team and everyone in the team cheers him up. This boosts his self-esteem and reinforces his belief in his importance and his contribution to the growth of his organization.
A collection of small gestures is what we spoke about and they hardly take 10 mins of a leader's time but the results from these gestures are exponential and well worth it.
Feedbacks help a manager in becoming a better leader. We have all seen and heard about the quote that 'a boss succeeds and good leaders lead people to success'. How will you lead your team to success if you don't know your team?
Feedbacks help in understanding the pulse of your team and in understanding the emotions of your team. Over a time the relationship builds between a leader and his/her team and in return, the leader starts working for the success of them and not him/her as an individual.
Also, some feedback may be about the management functions. Which can have two effects, can be turned against the employee who gave the feedback or can be used to improve the management and the manager themselves. This adds more value to the feedback and makes you a better leader.
Say, there is feedback from a set of employees saying the hiring process is old school and it needs a simple and effective plan in action. The managers can be little apprehensive about changing the process as it has been followed for years now but to keep up with the current generation and testing their skills in a right way, it makes sense to change the process step by progressive step.
On coming up with an effective plan, the manager will lead the hiring team into success as the candidates can be evaluated better and talented hires is an organisation's growth. Here the managers leads the team and the company to success.
These are just some of the core important factors that reflect the importance of feedback but they are not it. There is many more to it.
The voice of the employee adds value at a different level and the results, when implemented right, can prove that feedback is valuable and sensible.