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Ferentz optimistic on Iowa's vaccination rate
IOWA CITY – By the time the Iowa football season kicks off in September, coach Kirk Ferentz believes his team's COVID vaccination rate will have improved considerably.
Ferentz said Friday at the Hawkeyes' preseason media day that the number of players vaccinated for the coronavirus has grown steadily since he indicated last month at the Big Ten kickoff that Iowa's team vaccination rate stood at around 70 percent.
"That number has gone up," Ferentz said. "We continue to educate. That's our number one priority, to educate our players what the protocols are going to be and just what we all feel is probably in their best interest."
The Iowa coach, who has been vaccinated, repeated that he will not mandate that his players be vaccinated but he will continue to educate.
"When September rolls around, we expect to be a in pretty good position, but it remains our players choice what they choose to do," Ferentz said, encouraging fans to educate themselves about what the vaccination can do.
Ferentz said coaches have stressed to players that their individual choices can impact the entire team.
Vaccinated players are currently not subjected to contact tracing, which Ferentz said was at the root of multiple issues Iowa had within its team a year ago.
"What is different for us is we're all involved with a team activity," Ferentz said. "Whether you are a coach or a player, you only get limited opportunities. Players' windows are a lot smaller, pretty precious time and to give it up for something that is likely avoidable, that's a tough deal."
Back to work: A week into fall camp, quarterback Spencer Petras welcomes a return to a normal preseason routine.
"Especially after last year and how weird camp was, it's such a great feeling to be out here as part of a normal camp and putting the work in," Petras said. "I'm excited to be back at camp."
The junior quarterback has welcomed blending a number of newcomers into the mix as Iowa works toward a noon scrimmage Saturday that will be open to the public free of charge.
"It's been a great opportunity for our offense and it's been great for those guys coming in," Petras said. "It's been good for them to come in and compete right away. It's a lot of work, but it's been fun."
Still building: Iowa running back Tyler Goodson expects more of himself as a junior.
The Hawkeyes' rushing leader the past two years who was named the Big Ten running back of the year in 2020 sees room for growth in his game.
"When I watch tape, there are a lot of things that I can do better and that part of the work, it is never finished," Goodson said. "There are a lot of little ways to improve and getting those little things down, that's what I need to get done."
Significant start: Iowa's first two opponents, Indiana and Iowa State, are ranked in the top 15 in the preseason coaches poll, something Ferentz said creates a sense of urgency during fall camp.
"They're both really good teams and the margin for error is always tight. We traditionally have not always been as sharp early or at least where we need to be. Last year is a good example of that," Ferentz said, referencing the Hawkeyes' 0-2 start before winning its final six games.
"It's a good reminder for all of us that we don't have time to waste. Not every day is going to be perfect, but we need to be working hard."
Veteran group: It's been business as usual for Iowa's kicking unit.
A pair of sixth-year seniors, kicker Caleb Shudak and long snapper Austin Spiewak, are joined by holder Ryan Gersonde, a fifth-year senior, on one of the most veteran segments of the Hawkeye team.
"It's pretty unusual to have that much experience coming back," Shudak said. "But, it's something that can really help us out. We have to be close to being the most experienced part of the team. We've all been around forever."
Trading places: Freshman Griffin Liddle of Bettendorf has already made a move.
Listed as a defensive lineman on Iowa's preseason roster, Liddle is now working with the offensive linemen in fall camp.
Player Council named: Iowa announced its 16-player Player Council for the 2021 season on Friday, a group of six seniors and 10 juniors who will assist in formulating policies and team decisions.
Seniors Matt Hankins, Ivory Kelly-Martin, Jack Koerner, Kyler Schott, Caleb Shudak and Zach VanValkenburg were named to the group.
They were joined by juniors Dane Belton, Seth Benson, Jack Campbell, Joe Evans, Tyler Goodson, Sam LaPorta, Tyler Linderbaum, Kaevon Merriweather, Spencer Petras and Tyrone Tracy Jr.
Seats sold: Iowa officials announced Friday that around 5,000 seats remain for the Hawkeyes' Sept. 4 season opener against Indiana.
That number reflects an increase in season ticket sales at this point compared to the 2019 season and a sellout of Iowa's student season ticket allotment of 8,200.
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Write on the Money
Annual Student Writing Contest
Are you a Mid-State student who enjoys writing or has a strong example of a research paper or essay that you've already written? Submit your writing to compete for a $100 gift card and a chance to showcase your work!
This contest provides two categories for submission (Research Paper and Essay). Students can compete in one or both—the winner of each category will receive a $100 gift card from the location of their choice, donated by the Student Leadership Board.
Students are encouraged to submit existing writing that qualifies and adheres to the submission categories and guidelines, or write a new paper or essay just for the contest.
Qualifications for Entry
You must be currently enrolled as a student at Mid-State Technical College for the spring 2021 semester.
You may enter one paper per category.
Your paper must be original, unpublished nonfiction (no fiction or poetry).
Submission Categories
Research Paper: On any topic. Up to 2,500 words, double spaced, in APA style.
Essay: A descriptive, definition, narrative, personal, persuasive, opinion, comparison, classification/division, process analysis, or cause and effect essay. Up to 2,000 words, double spaced, in APA style.
Competing students will upload their writing submissions to Blackboard by 4:00 p.m. on Friday, March 5, 2021:
Log onto Blackboard and choose the Self-Enroll Courses tab on the top of the screen.
Choose the Special Events folder and choose Enroll in the drop-down menu.
Contest Timeline
Friday, March 5, 4:00 p.m. – Deadline for submission (contest entries due to Blackboard).
Friday, April 9, 4:00 p.m. – Contest winners notified.
This year's contest will be judged by acclaimed Wisconsin authors Nick White and Carrie Hagen.
Essay Category--Nick White, author of How to Survive a Summer; Research Paper Category--Carrie Hagen, author of We is Got Him
Nick White – Essay Judge
Originally from Mississippi, Nick White is the author of the novel How to Survive a Summer (Blue Rider/Penguin, 2017) and the story collection Sweet and Low. He is an assistant professor of English at The Ohio State University's MFA program in creative writing. His short stories, poems, and essays have appeared in a variety of places, including The Kenyon Review, Guernica, Catapult, The Hopkins Review, Indiana Review, The Literary Review, Lit Hub, and elsewhere. He is currently at work on a new novel.
Carrie Hagen – Research Paper Judge
Carrie Hagen is the author of We Is Got Him (Overlook, 2011), a narrative nonfiction account of the first recorded ransom kidnapping in American history. She lives and writes in Philadelphia, where she works in historic preservation and facilitates student writing workshops. A former high school English teacher and journalist, Hagen served as a research assistant to The Washington Post associate editor Steve Luxenberg on his book Annie's Ghosts (Hyperion, 2009) and as a consultant to Dr. David Kessler, former head of the FDA, on Captured: Unraveling the Mystery of Mental Suffering (HarperCollins, 2016). Other credits include delivering the keynote address at an FBI federal training in 2016 and participating as an expert commentator on the Travel Channel's Mysteries at the Museum. She holds an MFA in creative nonfiction from Goucher College.
Research category – Originality of ideas, clear expression of ideas, unified and cohesive structure, logically supported thesis, credibility of sources, synthesis of source material, and correct APA format.
Essay category – Originality of ideas, clear expression of ideas, unified and cohesive structure, well-supported thesis, and unique voice.
See the Writing Contest course in Blackboard for full details about the submission categories and guidelines, formatting requirements, etc.
See the Writing Contest course in Blackboard for more for detailed submission guidelines. If you still have questions, contact Jill Quinn, communication instructor, by phone at 715.422.5416 or email.

redpajama_set_name: RedPajamaCommonCrawl


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The New Zealand Investment Story
We promote the investment case of the asset class to local and offshore investors and encourage in their portfolio allocations to New Zealand Private Equity and Venture Capital.
New Zealand is strategically located in the Asia-Pacific region, with proximity and access to high-growth economies internationally.
Our workforce is talented, well-educated and competitively priced, with an aptitude for innovation and cutting edge technology, and a focus on excellence and quality.
We provide a politically and economically stable society, and a positive business environment that has one of the lowest levels of corruption in the world and is supported by a robust legal system that protects intellectual property.
New Zealand is recognised for a flair for original thinking and determined application, leading to the development of novel products and processes and unique resolutions for difficult problems.
We offer world-class research and development (R&D) capabilities, based on a collaborative relationship between research institutes, universities and the private sector, and supported by government initiatives and investment.
The New Zealand Private Equity Market
This report provides an overview of the New Zealand private equity market, with specific focus on the institutional funds management industry. It examines the key features of the market relevant to institutional investors – the size of the market, investment activity, the fund manager landscape and historical returns.
Doing Business in New Zealand
Doing Business in New Zealand Guide introduces you to New Zealand's business and trading environment, with particular focus on legal and regulatory matters.
The Investors Guide to the New Zealand Beverage Industry 2017
The New Zealand wine industry has achieved phenomenal growth, producing world renowned wine varieties.
Outside wine, New Zealand juice, beer and alcoholic spirits are all showing significant potential.
The Investors Guide to the New Zealand Meat Industry 2017
New Zealand is the global leader in lamb and deer meat exports and has a strong position in beef exports; other species are primarily focused on the domestic market, although in recent years chicken exports have been growing.
The Investors Guide to the New Zealand Diary Industry 2017
New Zealand is a global leader in dairy trade, gaining export market share in the past decade.
New Zealand is the eighth largest milk producing country in the world and accounts for 3 per cent of global milk production. New Zealand is a global leader in the exporting of powders, caseins, butter/dairy fats, and powders, with a strong position in cheese and lactose.
The Investors Guide to the New Zealand Processed Food Industry 2017
New Zealand has a rapidly growing processed foods sector, leveraging on available raw and unique materials.
New products, exports and firms continue to emerge, though many of these firms require significant capital to fund growth.
The Investors Guide to the New Zealand Produce Industry 2017
New Zealand is a major producer and exporter of kiwifruit and apples, accounting for over 70 per cent of the country's fresh produce export value.
New Zealand also produces a wide range of other fruit and nuts, with significant growth in avocados, cherries and blueberries. We are a mid-sized producer of vegetables for processing and export.
The Investors Guide to the New Zealand Seafood Industry 2017
New Zealand has the tenth longest coastline of any country in the world, access to a strong set of natural resources, and exports seafood to 112 countries.
New Zealand has an efficient, modern seafood industry and large and sustainably managed wild catch fishery. Aquaculture showing huge theoretical growth potential for New Zealand and is based around three species; mussels, salmon, and oysters.
New Zealand Space Economy: Its value, scope and structure
Executive Summary New Zealand is a unique example of a space economy almost entirely driven by commercial activity The Ministry…
Honey and Software Win NZ Private Capital Investment of the Year
24 October 2019 Honey and Software Win NZ Private Capital Investment of the Year Successful investments by two private equity…
The offshore perspective on investment into New Zealand. Simpson Grierson's latest report, produced in association with Mergermarket, is based on…
The Do's and Don'ts of Private Capital for Entrepreneurs
Here are four things you should do, and four you shouldn't, when it comes to private capital. Here's a hard-earned…
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Immune signatures for HIV-1 and HIV-2 induced CD4+T cell dysregulation in an Indian cohort
Sukeshani Salwe1,
Amitkumar Singh1,
Varsha Padwal1,
Shilpa Velhal1,
Vidya Nagar2,
Priya Patil2,
Alaka Deshpande2 &
Vainav Patel ORCID: orcid.org/0000-0001-6661-32381
HIV-2 infection is characterised by a longer asymptomatic phase and slower AIDS progression than HIV-1 infection. Identifying unique immune signatures associated with HIV-2 pathogenesis may thus provide therapeutically useful insight into the management of HIV infection. This study examined the dynamics of the CD4+T cell compartment, critical in disease progression, focussing on chronic HIV-2 and HIV-1 infected individuals at various stages of disease progression.
A total of 111 participants including untreated and treated HIV infected individuals and seronegative individuals were enrolled in this study. The relative proportion of CD4+T cell subsets, expressing CD25 (IL-2Rα) and CD127 (IL-7R), in HIV infected individuals and seronegative controls were assessed by multiparametric flow cytometry. Additionally, levels of immune activation and cytotoxic T lymphocytes in both the CD4+T and CD8+T cell compartments was evaluated.
Both treated and untreated, HIV-1 and HIV-2 infected individuals showed apparent dysregulation in CD4+ T cell subset frequency that was associated with disease progression. Furthermore, longitudinal sampling from a group of HIV-1 infected individuals on virologically effective ART showed no significant change in dysregulated CD4+T cell subset frequency. For both ART naïve and receiving groups associations with disease progression were strongest and significant with CD4+ T cell subset frequency compared to per cell expression of IL-2Rα and IL-7Rα. In untreated HIV-2 infected individuals, T cell activation was lower compared to ART naïve HIV-1 infected individuals and higher than seronegative individuals. Also, the level of Granzyme-B expressing circulating T cells was higher in both ART-naïve HIV-1 and HIV-2 infected individuals compared to seronegative controls.
Dysregulation of IL-2 and IL-7 homeostasis persists in CD4+T cell subsets irrespective of presence or absence of viremia or antiretroviral therapy in HIV infection. Furthermore, we report for the first time on levels of circulating Granzyme-B expressing CD4+T and CD8+T cells in chronic HIV-2 infection. Lower immune activation in these individuals indicates that persistent immune activation driven CD4+T cell depletion, as observed in untreated HIV-1 infected individuals, may not be as severe and provides evidence for a disparate pathogenesis mechanism. Our work also supports novel immunomodulatory therapeutic strategies for both HIV-1 and HIV-2 infection.
An estimated total of 36.7 million [34.0 million–39.8 million] individuals worldwide, are people living with human immunodeficiency virus (PLHIV) [1].Of these, around 2.1 million [1.71 million–2.64 million] are estimated to live in India [2].The HIV epidemic in India largely involves sexual transmission of HIV-1 Clade C viruses [3]. Furthermore, HIV-2 single infection and dual (HIV-1 along with HIV-2) infection is clearly present and persistent in the HIV infected population in India [4, 5]. Both HIV-1 and HIV-2 share the same modes of transmission, namely sexual contact, blood-borne exposure (blood transfusion, shared needles) and perinatal transmission. Also, both infections are associated with a progressive decline of CD4+T cells and loss of immune function, which manifests clinically as an increased susceptibility to opportunistic infections. However, the pathogenesis associated with these two viruses is reported to be disparate. HIV-2 infection has been associated with a significantly longer asymptomatic stage, lower plasma viral load, slower decline in CD4+T-cell count, and lower mortality rate attributable to AIDS compared to HIV-1 [6,7,8]. In addition, a well-preserved and polyfunctional HIV-specific memory CD4+T cell response has been shown to be a hall mark of HIV-2 infection [9, 10].
Thus delineating unique immune signatures associated with HIV-2 pathogenesis may provide therapeutically useful insight into the management of HIV-1 infection and AIDS. The CD4+T cell compartment is critical in disease progression as it serves both, as a target and also as a marker of disease progression. Also, CD4+ T cell help is an important component of immune responses, both cellular and humoral. In this study, we examined the dynamics of the CD4+ T cell compartment in terms of various important subsets, based on homeostatic markers, focussing on chronic HIV-1 and HIV-2 infected individuals at various stages of disease progression.
To address homeostatic turnover, CD4+T cell subsets have been defined with respect to expression of Interleukin-2 (IL-2) and Interleukin-7 (IL-7) receptors [11]. The receptors for IL-2 and IL-7 share the common γ-chain in combination with unique high-affinity receptors: IL-2Rα (CD25) or IL-2Rβ (CD122) in the case of IL-2 and IL-7Rα (CD127) in that for IL-7 [12]. IL-2 and IL-7 function through their receptors as master regulators of T cell development, naïve and memory T cell homeostasis, proliferation, and differentiation [12,13,14]. CD4+ T cells can be categorized into three subsets on the basis of expression of IL2Rα (CD25) and IL7R (CD127): CD127+CD25low/−, CD127−CD25−, and CD25highCD127low as previously reported [11]. This report characterized these phenotypes for the state of differentiation (i.e., naive vs. central memory vs. effector memory) using CD45RA and CD62L in HIV-1 and seronegative individuals. It was observed that CD127+CD25low/− subset corresponded to both CD45RA+CD62L+ naive cells and CD45RA− CD62L+ central memory cells. The CD127− CD25− subset consisted of mainly CD45RA−CD62L− effector memory cells. The CD25highCD127low phenotype of Regulatory T cells (Treg) has been defined by various study groups [15, 16] and was validated to correspond to CD4+CD25+Foxp3+ Treg cells also by us (Additional file 1: Figure S1). As, the dynamics of the CD4+T cell compartment may differ based on the presence or absence of active viral replication, we have evaluated the relative proportions of three subsets of CD4+ T cells that are defined based on expression of CD127 and CD25 under anti-retroviral therapy (ART) naïve and therapy receiving conditions in both HIV-1 and HIV-2 infected individuals. Furthermore, some HIV-1 infected individuals were followed longitudinally to assess the effect of long term ART on the dynamics of CD4+T cell compartment.
Systemic chronic immune activation is considered as the driving force of CD4+T cell depletion and resulting disease progression following HIV infection [17, 18], making it an important cellular parameter to evaluate in the context of HIV-1 and HIV-2 infection. Also, cytotoxic T cells (CTL) expressing the effector molecule Granzyme B (GrzB) are central in conferring host cell immunity against viral pathogens through recognition and killing of infected cells [19,20,21,22]. Thus, this study was designed, to examine levels of activation as well as Granzyme B expression in the both CD4+T and CD8+ T cells in chronic HIV infection and to delineate any unique immune signatures specific to either HIV-1 or HIV-2 infection.
A total 111 individuals were recruited for this cross sectional study including 33 seronegative, 24 antiretroviral therapy (ART) naïve HIV-1infected individuals, 16 ART-naïve HIV-2 infected individuals,19 ART-receiving HIV-1 and 19 ART- receiving HIV-2. All the participants were recruited from ART Centres, in Maharashtra with approval from the NIRRH Institutional Clinical Ethics Committee (Project no.:225/2012) and informed consent was obtained from all of them. The clinical features of the enrolled participants (age, viral load, absolute CD4 count and status of antiretroviral therapy) are shown in Table 1. Total nucleic acid from blood was isolated using the MagNa pure Compact Nucleic Acid Automated System (Roche Diagnostic, Germany) and plasma viral load of HIV-1 infected individuals was estimated using Cobas TaqMan Real Time PCR (Roche Molecular Systems, Piscataway, NJ) with a limit of detection of 34 RNA copies/ml. Due to the non-availability of a standard, reproducible HIV-2 viral load estimation assay and thus the inability to compare viral loads from HIV-1 infected individuals, we were unable to estimate the viral load of HIV-2 infected individuals.
Table 1 Clinical characteristics of participants
Immunophenotypic analysis of T cell subsets
For immunophenotypic staining peripheral blood collected in EDTA vacutainer were stained with appropriate fluorochrome-conjugated surface antibodies, including anti-CD3 (Clone:SK7), anti-CD4 (Clone:RPA-T4), anti-CD8 (Clone:SK1), anti-CD25 (Clone:M-A251), anti-CD127 (Clone: HIL-7R-M21), anti-HLADR (Clone:L243), anti-CD38 (Clone:HIT2), anti-CD45RA (Clone:HI100) and anti-granzyme (Clone:GB11); purchased from either BD Biosciences or Biolegend. Intracellular staining for Granzyme was performed according to manufacturer's instructions (BD Cytofix/Cytoperm™ Plus, Catalog No.-554,715) after surface staining with specific surface marker antibodies. The samples were processed on the same day of sampling for ex-vivo staining and ICCS Assay for Granzyme detection. Flow cytometric acquisition and analysis were performed on at least 50,000 acquired events (gated on lymphocytes) on a BD ACCURI C6 flow cytometer (BD Biosciences). The 670LP and 675/25 filters were used to measure the fluorescence corresponding to anti-CD25 and anti-CD127 antibodies respectively. The stochastic % standard deviation (SD) of MFI for 670LP and 675/25 filter was calculated using Spherotech 6-Peak (Cat No-653145, BD Biosciences) and 8-peak (Cat No-653144, BD Biosciences) Validation Beads and was found to be 3.78 and 2.43% respectively for the period during which study samples were acquired. Data Analysis was performed using FlowJo (Tree Star Inc., Ashland, Oregon, USA).
Statistical analysis was performed using GraphPad Prism version 5.00 (GraphPad Software, San Diego, California, USA). The data are presented as scatter plots, with bars indicating median values and groups were compared using unpaired t-test with Welch's correction 95% confidence interval. The prospective data was analysed using Repeated measures ANOVA and non-parametric paired T test (Wilcoxon matched). Non parametric Spearman's correlation coefficient was used to assess the correlation between two variables. P values less than 0.05 were considered significant.
Distribution of CD4+T cell subsets defined on the basis of expression of CD127 (IL-7R) and CD25 (IL-2Rα) in both HIV-1 and HIV-2 infected ART-naïve individuals
When the relative proportions of these CD4+T cell subsets were examined in ART-naïve HIV-1 and HIV-2 infected individuals, we observed a significant increase in the frequency of the Tregs (CD25highCD127low) subset (P = < 0.0001 for HIV-1; P = < 0.0001 for HIV-2) and effector memory (CD127−CD25−) subset (P = < 0.0001 for HIV-1; P = < 0.0001 for HIV-2), and a decline in the fraction of naive/central memory (CD127+CD25low/−) T cell subset (P = < 0.0001 for HIV-1; P = < 0.0001 for HIV-2) in both HIV-1 and HIV-2 infected individuals as compared to seronegative controls. Also, the frequency of these CD4+T cell subsets was found to be similar in both ART-naïve HIV-1 and HIV-2 infected individuals (Fig. 1).
Identification of dysregulation in CD4+T cell subsets based on the expression of CD127 (IL-7R) and CD25 (IL-2Rα). a Gating strategy for defining subsets of CD4+ T cells using CD127 and CD25. Cells were gated based on characteristic light scatter properties FSC against SSC, followed by gating on CD4+ T cells. Thereafter based on expression of CD127 and CD25, CD4+T cells were further demarcated as naive/memory (CD127+CD25low/−), effector (CD127−CD25−) and Tregs (CD25highCD127low). b Comparison of frequency of CD4+ T cells subsets in ART-naïve HIV-1 (n = 21), HIV-2 (n = 16) infected individuals and seronegative individuals (n = 33). Statistical significance was evaluated by unpaired t test; *, p < 0.05; **, p < 0.01; and ***, p < 0.001
To assess the association of these CD4+T cell subsets with disease progression under ART-naïve conditions in both HIV-1 and HIV-2 infected individuals, we correlated the frequency of these subsets with absolute CD4 count. Tregs (CD25highCD127low) subsets and effector memory (CD127−CD25-) cells were found to be negatively correlated and naive/memory (CD127+CD25low/−) T cell subset was found to be positively correlated with absolute CD4 count in both HIV-1 and HIV-2 study groups (Table 2). In addition, the degree of expression per-cell was estimated using median fluorescence intensity (MFI) for both CD25 and CD127 on CD4+T cell subsets and compared across groups. No significant difference was observed in the MFI of either of these markers in CD4+T cell subsets between HIV infected individuals (MFI of CD25: P = 0.1537 for HIV-1; P = 0.1380 for HIV-2, MFI of CD127: P = 0.1345 for HIV-1; P = 0.1012 for HIV-2) and seronegative controls (Fig. 4b). Furthermore, when the MFI for both markers was correlated with absolute CD4 count, no significant correlation with disease progression was observed (Table 2). To address any effect of unequal sample size between seronegative group and HIV infected groups on statistical analysis, a bootstrapping analysis was carried out (Additional file 2: Figure S2) which showed the same observations as reported above. Also to address any effect of unmatched ages of participant groups on statistical analysis, age matching was performed (Additional file 3: Figure S3).This had no impact on reported observations.
Table 2 Association of CD4 T cell subsets, MFI of CD25 and CD127, Level of immune activation and level of cytotoxicity with absolute CD4 count across the groups
Immune activation, under ART-naïve conditions, in both HIV-1 and HIV-2 infection
In our study, the frequency of activated (CD38+HLA-DR+) cells in both CD4+T (P = 0.0027 for HIV-1; P = 0.0119 for HIV-2) and CD8+T cell (P = < 0.0001 for HIV-1; P = 0.0071 for HIV-2) compartments was found to be elevated in both HIV-1 and HIV-2 infected groups as compared to seronegative controls. We also observed elevated levels of activation (CD38+HLA-DR+ cells) in HIV-1 infected individuals as compared to HIV-2 infected individuals in both CD4+T (P = 0.0117) and CD8+T cell (P = 0.0022) compartments (Fig. 2a). The elevated level of activation (CD38+HLA-DR+ cells) in the CD4+T and CD8+T cell compartment negatively correlated with absolute CD4 count in HIV-1 infected individuals, but not with that in HIV-2 infected participants (Table 2).
Level of Immune activation in HIV-1 and HIV-2 chronic infection. a Comparison of expression of immune activation markers (CD38&HLA-DR) on CD4+T and CD8+T cells among ART-naïve HIV-1 (n = 11), HIV-2 (n = 13) infected individuals and seronegative individuals (n = 11). Statistical significance was evaluated by unpaired t test; *, p < 0.05; **, p < 0.01; and ***, p < 0.001. The frequencies of activated cells are those observed within CD4+T and CD8+T cell gates as outlined in Additional file 4 Figure S4. b Correlation analysis between frequency of CD4+ and CD8+ activated T cells and Tregs cells in both ART-naïve HIV-1(n = 11) and HIV-2 (n = 13) infected individuals. Spearman's correlation test was used to determine the correlation coefficient (r) and the significance (p < 0.05).
Recent studies have suggested that Tregs, might play a role in the dynamics of immune activation during HIV-1 infection [23,24,25]. However no study has assessed relationship between Tregs and the activation (CD38+HLA-DR+ cells) status of CD4+ or CD8+T cells in HIV-2 infection. On performing this analysis, we observed a positive correlation (though not significant) between the proportions of activated CD4+T cells and Tregs in both HIV-1 (r = 0.5030, P = 0.1383) and HIV-2 infected (r = 0.4594, P = 1142) study groups. Interestingly, a clear and significant positive correlation was observed between activated CD8+T cells and Tregs among HIV-1 infected individuals (r = 0.6364, P = 0.0479) and not in those infected with HIV-2 (r = 0.4704, P = 0.1047) (Fig. 2b).
Granzyme-B expression in CD4+ and CD8+T cells in ART-naïve infected individuals
We observed that, the level of GrzB expression in CD4+T (P = 0.0138 for HIV-1; P = 0.0355 for HIV-2) and CD8+T cells (P = < 0.0001 for HIV-1; P = 0.0003 for HIV-2) was substantially higher in both HIV-1 and HIV-2 infected individuals compared to seronegative controls. However, the level of GrzB expression in CD4+T (P = 0.8150) and CD8+T cells (P = 0.0673) was found to be similar in both HIV-1 and HIV-2 infected individuals (Fig. 3a).
Expression of Granzyme B in CD4+T and CD8+T cells. a Comparison level of GrzB from CD4+ T and CD8+T cells among ART-naïve HIV-1(n = 14), HIV-2 (n = 13) infected individuals and seronegative individuals (n = 14). Statistical significance was evaluated by unpaired t test; *, p < 0.05; **, p < 0.01; and ***, p < 0.001. Gating strategy outlined in Additional file 4: Figure S4. b Correlation analysis between level of GrzB and frequency of Tregs cells in both ART-naïve HIV-1 (n = 10) and HIV-2 (n = 13) infected individuals.Spearman's correlation test was used to determine the correlation coefficient (r) and the significance (p < 0.05)
To investigate whether levels of total circulating cytotoxic T cells were associated with disease progression, the correlation between levels of GrzB expressing cells within the CD4+T and CD8+T cell subsets with absolute CD4 count was carried out. As shown in Table 2, a significant positive correlation was observed between levels of GrzB expressing CD4+T cells and absolute CD4 count in HIV-1 infected individuals. Additionally, a significant negative correlation between levels of GrzB expressing CD8+T cell and absolute CD4 count in HIV-1infected individuals was observed. However this correlation was not observed in HIV-2 infected individuals. Furthermore, when we assessed the association of Tregs and cytoxicity, as expected, a significant negative correlation (r = − 0.6636, P = 0.0260) was detected between frequency of GrzB producing CD4+T cells and frequency of Tregs in HIV-1 infected study group (Fig. 3b). Also, positive correlation was detected between frequency of GrzB producing CD8+T cells and frequency of Tregs in the HIV-1 infected study group (r = 0.5831, P = 0.0597). However, no significant association was detected between the level of cytotoxicity and Tregs in HIV-2 infected individuals (r = 0.0962, P = 0.7543 for GrzB+CD4+T cells; r = 0.2779 P = 0.3580 GrzB+CD8+T cells).
Effect of antiretroviral therapy on CD4+T cell subset defined on the basis of expression of CD25 and CD127
We sought to determine the frequency of CD4+T cell subsets in the whole blood of individuals who were on extended (> 1 year), successful (HIV-1 viral load < 34copies/ml) uninterrupted ART in comparison to ART-naïve individuals and seronegative individuals.
We observed no alteration in the frequency of the Tregs (CD25highCD127low) subset, effector memory (CD127−CD25−) subset and naive/memory (CD127+CD25low/−) T cell subset between ART- receiving and ART-naïve HIV-1 and HIV-2 infected individuals. However, we observed significantly increased levels, similar to those observed in ART-naïve infected individuals, in the frequency of the Tregs (CD25highCD127low) subset (P = < 0.0001 for HIV-1; P = < 0.0001 for HIV-2), effector memory (CD127−CD25−) subset (P = < 0.0001 for HIV-1; P = 0.0002 for HIV-2), and decline in the fraction of naive/memory (CD127+CD25low/−) T cell subset (P = < 0.0001 for HIV-1; P = < 0.0001 for HIV-2) compared to seronegative controls (Fig. 4a). When the MFI of CD25 within Tregs was evaluated, we observed significant reduction in ART- receiving HIV infected individuals (P = 0.0002 for HIV-1; P = 0.0454 for HIV-2) compared to that in ART-naïve individuals. Moreover, the MFI of CD127 within naive/memory (CD127+CD25low/−) T cell subset was also observed to be significantly lower in ART- receiving HIV-2 infected individuals (P = 0.0007) compared to ART-naïve HIV-2 infected individuals (Fig. 4b). A similar trend (p = 0.06) was observed for HIV-1 infected individuals. The % reduction in MFI of CD25 within Tregs was 38 and 27.6% for HIV-1 and HIV-2 infected individuals respectively when compared to the MFI of corresponding ART-naïve groups. Also the % reduction in MFI of CD127 within naive/memory (CD127+CD25low/−) T cell subset was 17.9% for ART- receiving HIV-2 infected individuals compared to ART-naïve group. These changes in MFI were clearly higher than stochastic variation of fluorescence intensity calculated for the instrument during the period of study (see methods).
Effect of Antiretroviral therapy on CD4+T cell subset defined on the basis of expression of CD25 and CD127. a Comparison of CD4+ T cells subsets in ART-naïve HIV-1 (n = 21), ART-treated HIV-1 (n = 19), ART-naïve HIV-2 (n = 16), ART-treated HIV-2 (n = 19) infected individuals and seronegative individuals (n = 33). b Comparison of MFI of CD25 and CD127 in ART-naïve HIV-1 (n = 21), ART-treated HIV-1 (n = 19), ART-naïve HIV-2 (n = 16), ART-treated HIV-2 (n = 19) infected individuals and seronegative individuals (n = 33). Statistical significance was evaluated by unpaired t test; *, p < 0.05; **, p < 0.01; and ***, p < 0.001
When the association between absolute CD4 count and frequency of CD4+T subsets in ART- receiving HIV-1 and HIV-2 infected individuals was examined, a significant negative correlation was observed between the absolute CD4 count and the frequency of the Tregs (CD25highCD127low) and a significant positive correlation was observed between the absolute CD4 count and the frequency of the naive/memory (CD127+CD25low/−) T cell subset in ART- receiving HIV-2 infected individuals (Table 2). No correlation was observed between absolute CD4 count and frequency of CD4+T cell subsets in ART- receiving HIV-1 infected individuals. Also the MFI of CD127 within naive/memory (CD127+CD25low/−) T cell subset and MFI of CD25 within Tregs in ART- receiving HIV-1 infected individuals showed no correlation with absolute CD4 count. However MFI of CD127 within naive/memory (CD127+CD25low/−) T cell subset showed significant positive correlation with absolute CD4 count in ART- receiving HIV-2 infected individuals. No correlation was observed between absolute CD4 count and MFI of CD25 within Tregs in ART- receiving HIV-2 infected individuals (Table 2).
Further, we followed 10 ART-receiving (for more than one year) HIV-1 infected individuals at 3 months (TP-2) and 18 months (TP-3) after enrolment (TP-1) in this study to evaluate whether extended ART was able to restore the dysregulated CD4+Tcell subset frequencies to normal levels. Due to difficulties in follow up, we were unable to follow all individuals at each time point. As presented in Table 3A, CD4+T cell subset data was available for TP-1 and TP-2 from 8 individuals (Group-1), for TP-1 and TP-3 from 2 individuals (Group-2) and for TP-1, TP-2 and TP-3 from 4 individuals (Group-3). Assessment of the relative proportions of these three CD4+T cell subsets was carried out for Group-1 and Group-3, where as shown in Table 3B and C respectively, no significant change was observed. Figure 5 represents the graphical presentation of the data.
Table 3 Prospective data analysis of CD4 T cell subsets
Prospective data analysis of CD4+T cell subsets. Graphical presentation of CD4+T cell subset frequencies from 10 ART-receiving (for more than one year) HIV-1 infected individuals at 3 months (TP-2) and 18 months (TP-3) after enrolment (TP-1). Bold rectangular line represents the mean ± SEM frequency of corresponding CD4+ T cell subset in seronegative individuals (n = 33)
This is the first study evaluating concurrently the dynamics of the T cell compartment using homeostatic markers such as CD25 (IL-2Rα) and CD127 (IL-7R), level of immune activation and circulating cytotoxic T cell levels in HIV-2 infected individuals. Furthermore, the dynamics, in terms of these markers of the CD4+T cell compartment has been addressed separately in the absence of viremia due to effective ART in individuals infected with HIV-1 or HIV-2 in the Indian population for the first time.
Progressive decline of CD4+T cells following HIV infection is a hallmark of disease progression resulting in HIV associated immune dysfunction [26]. In this study, both ART-naïve HIV-1 and HIV-2 infected individuals showed apparent dysregulation (altered CD25 and CD127 expression) in CD4+T cell subsets and this dysregulation was found to be associated with disease progression. These results suggest an altered cytokine network with respect to IL-2 and IL-7 homeostasis present in HIV infected individuals. Also, the increased frequency of Tregs (CD25highCD127low) and effector memory (CD127−CD25−) subsets seen in both infections within this study may reflect an increased level of immune activation present in these individuals as consequence of viremia in the absence of ART. The significantly lower frequency of the naive/memory (CD127+CD25low/−) T cell subset is congruent with loss of central memory CCR5 expressing CD4+T cells as they represent the primary target of HIV infection. Our results with respect to increased frequency of Tregs in ART-naïve HIV infected individuals compared to seronegative individuals are in agreement with some previous reports describing HIV-1 clade B and HIV-2 clade A infection [24, 27,28,29]. However, our result with respect to increased frequency of Tregs in ART-naïve HIV-1 infected individuals are not in agreement with some reports [11, 27] that used PBMC staining and were unable to detect differences in frequency of Tregs defined as CD4+CD25bright [27] and CD25highCD127low [11] compared to seronegative individuals.
Effective Antiretroviral therapy (ART) blunts viral replication and should result in an increase of the CD4+T cell count in peripheral blood, a phenomenon known as immune reconstitution [30]. However the data available so far is limited and conflicting with respect to observed differences in the frequency of Tregs in HIV-1 infection followed by extensive ART [30,31,32,33,34]. No study has assessed the effect of ART on CD4+T subset defined on the basis of expression of CD25 and CD127 in HIV-1 and HIV-2 infection. Analysing the effect of ART on these CD4+T cell subsets revealed that ART-receiving HIV infected individuals showed similar frequencies of circulating CD4+T cell subsets to those in ART-naïve HIV infected individuals and were higher than seronegative individuals. Also longitudinal assessment of these CD4+T cell subsets in ART- receiving HIV-1 infected individuals revealed that they were maintained at high levels over an extended period of ART in the absence of detectable viral replication. These results suggest that ART was not able to rectify a disruption which was occurring in immune homeostasis with respect to IL-2Rα (CD25) and IL-7R (CD127) expressing CD4+T cells in both HIV-1 and HIV-2 infection. Though, we recognise that this data would be stronger if it included baseline frequency value of these subsets, which was not available, before the initiation of ART.
Median fluorescent intensity is 'per-cell' basis expression of a marker on a cell and alteration in MFI's could also indicate the functional state for that marker. This paradigm has been supported by studies reporting CD25 expression and functionality of Treg cells [35]. Thus, we evaluated the MFI of both CD127 as well as CD25 in CD4+T cell subsets to ascertain if these correlated with disease progression as was found to be the case for frequencies of the subsets that are defined by them. The MFI of CD127 within naive/memory (CD127+CD25low/−)T cell subset and MFI of CD25 within Tregs (CD25highCD127low) in ART-naïve HIV infected individuals showed no association with disease progression which suggests that frequency of CD4+ T cell subsets defined by their expression are better predictors of disease progression compared to their per cell expression on these subsets.
Though there were no differences in the frequency of CD4+T cell subsets in ART- receiving HIV infected individuals compared to ART-naïve HIV infected individuals, we observed reduced per cell expression of CD25 and CD127 in ART- receiving HIV infected individuals compared to ART-naïve HIV infected individuals. This suggests that ART was able to alter the expression of IL-2Rα and IL-7R following successful virological control. While this is indicative of possible or impending immune restoration, it is noteworthy that it was the frequency of CD4+ T cell subsets and not MFI of CD25 or CD127 within these subsets that continued to show stronger correlation with disease progression (absolute CD4 count) in HIV infected individuals undergoing virologically effective ART.
Overall, we showed that dysregulation of CD4+T cell subsets occurred in infected individuals irrespective of the presence or absence of ART suggesting that the relative disruption in cytokine networks with respect to IL-2 and IL-7 homeostasis caused by HIV infection is persistent in infected individuals and may not be rectifiable through ART alone. From a clinical management point of view in India, our work provides the rationale for exploring new therapeutic strategies for both HIV-1 and HIV-2 infection aimed at directly correcting the HIV-induced immune dysfunction. Also both IL-2 and IL-7 are being investigated as therapies to improve the immune system function in HIV-infected individuals [36,37,38,39].
Systemic immune activation is a feature of chronic HIV infection and determines the rate of disease progression [17, 18, 40]. HIV-2 infection has been reported to be associated with slower disease progression as compared to HIV-1.However the data available with respect to immune activation associated with infection is limited, conflicting and has reported on either CD4 or CD8 compartment in isolation [41,42,43,44]. The present study explored the level of immune activation in ART-naïve HIV-1 and HIV-2 infection in CD4+T and CD8+T cells.
Our findings demonstrate that in ART-naïve HIV-2 infected individuals, the level of immune activation in both CD4+T and CD8+ T cell compartment was lower compared to ART-naïve HIV-1 infected individuals and higher than seronegative individuals. While lower levels of immune activation could possibly be explained, in part due to lower viral load as has been reported in previously [41, 45], it is important to note that in our study, the levels of circulating Tregs were observed to be similar to those in ART-naïve HIV-1 infected individuals. Furthermore, these levels correlated inversely with absolute CD4 counts. Thus, these results suggest that the activation induced Treg response following both HIV-1 and HIV-2 infection while similar in magnitude may be more effective in ART-naïve HIV-2 infected individuals. This would explain the lower levels of immune activation as well as higher absolute CD4 counts observed and hence contribute to better disease progression. Interestingly, we also observed that the level of immune activation in CD4+T and CD8+T cell compartment was not associated with disease progression (absolute CD4 count) in ART-naïve HIV-2 infected study group in contrast to ART-naïve HIV-1 infected individuals. This data indicates that the observed lower levels of immune activation in ART-naïve HIV-2 infected individuals were not sufficiently high enough to drive persistent immune activation driven CD4+T cell depletion as observed in ART naïve HIV-1 infected individuals.
Release of Granzyme B, the principle Granzyme in CD8+cytotoxic T lymphocyte (CTL) and accompanying cytolytic destruction of target cells is believed to be one of the key immunological mechanisms for destruction of virally infected cells including those infected by HIV [46]. Also, with respect to HIV-1 and SIV infection, studies have delineated a role for CD4+ T cytotoxic responses in the prevention of acquisition, peak and set point viremia, control of rebounding virus in the absence SIV specific CD8+T cells and in effective CD8+T cell mediated responses against infected cells [47,48,49,50].
We report on the level of Granzyme-B expressing circulating CD4+T and CD8+T cells in ART-naïve HIV-2 infected individuals which, as in the case of ART-naïve HIV-1 infected individuals was observed to be higher compared to seronegative controls. These cytotoxic CD4+T and CD8+T cell may be assumed to include the HIV-specific cytotoxic T cells and indeed, correlation analysis between Granzyme-B expressing circulating CD4+T and CD8+T cells with absolute CD4 count showed a significant, though opposing correlation in HIV-1 infected individuals, suggesting that Granzyme-B expressing circulating CD4+T cells may be playing a protective role against disease progression in HIV-1 infection. Also, as expected, these CD4+T cell Granzyme-B responses correlated negatively with Treg frequencies in ART-naïve HIV-1 infected individuals. Intriguingly, no such correlation of Granzyme-B producing T cells with Treg frequency (or absolute CD4 count) and thus disease progression was observed in ART- naïve HIV-2 infected individuals. Taken together with the lack of correlation of activation levels and absolute CD4 counts in ART-naïve HIV-2 infected individuals, this data suggests a disparate pathogenesis for HIV-2 where possibly lower viral load and efficient Treg activity are associated with disease progression rather than activation and cytotoxic function of T cells. A future study, focusing on virus specific cytotoxic T cell responses would definitely provide a clearer picture.
The study reports for the first time on levels of circulating Granzyme-B expressing CD4+T and CD8+T cells in chronic HIV-2 infection. Our data also highlights a possibly distinct pathogenesis mechanism operating in HIV-2 infected individuals who, despite having similar duration of infection have possibly lower viral load and efficient Treg activity. This would contribute to less activation driven CD4+T cell depletion and thus better disease progression. Furthermore, the observed disruption in cytokine networks with respect to IL-2 and IL-7 homeostasis caused by HIV infection is persistent in infected individuals and may not be rectifiable through ART alone. Our work also provides the rationale for exploring novel immunomodulatory therapeutic strategies for both HIV-1 and HIV-2 infection aimed at directly correcting the HIV-induced immune dysfunction.
Anti-retroviral therapy
CTL:
Cytotoxic T cells
GrzB:
Granzyme B
IL-2:
MFI:
Median fluorescence intensity
PLHIV:
People living with human immunodeficiency virus
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We are grateful to Dr. Paul Wallace (Director, Flow and Image Cytometry Facility Roswell Park Cancer Institute, Buffalo, NY, USA) for discussions pertaining to organisation and presentation of the manuscript. We also wish to acknowledge the assistance of Mr. Vaibhav Shinde in the preparation of figures for the manuscript. We extend our gratitude to Dr. Shahina Begum for assistance during statistical analysis.
This study was funded by Department of Biotechnology (DBT), India with grant no. (BT/PR6202/GBP/27/383/2012) awarded to the corresponding author. Ms. Sukeshani Salwe is recipient of fellowship from the University Grants Commission (UGC), India (201314-RGNF-2013-14-SC-MAH-37835) under Rajiv Gandhi National fellowship (RGNF) programme. The funding agency, DBT, evaluated the scientific merit of the grant under which this study was implemented and played no direct role in design of the study, sample collection, analysis, and interpretation of data or in writing the manuscript.
All raw data are available by request to corresponding author.
Department of Biochemistry and Virology, National Institute for Research in Reproductive Health, Indian Council of Medical Research, J. M. Street, Parel, Mumbai, 400012, India
Sukeshani Salwe, Amitkumar Singh, Varsha Padwal, Shilpa Velhal & Vainav Patel
Department of Medicine, Grant Medical College & Sir J. J. group of Hospitals, Mumbai, 400008, India
Vidya Nagar, Priya Patil & Alaka Deshpande
Sukeshani Salwe
Amitkumar Singh
Varsha Padwal
Shilpa Velhal
Vidya Nagar
Priya Patil
Alaka Deshpande
Vainav Patel
VP2, SS: Designed the study and analysed the data; SS, AKS, VP1, SV: Recruited the participants and performed the experiments; VN, PP, AD: Assisted with participants recruitment & obtaining clinical history; VP2, SS: Wrote the manuscript. All authors read and approved the final manuscript.
Correspondence to Vainav Patel.
Written informed consent was obtained from all participants, and the study was performed in accordance with the ethical guidelines and approved by NIRRH Institutional Clinical Ethics Committee (Project no.:225/2012).
Figure S1. Validation CD4+CD25highCD127low phenotype as Tregs. The lymphocyte population (A) was gated, followed by gating on CD4+ cells (B). Thereafter CD4+ lymphocytes, based on expression of CD25, CD127 and FOXP3 were demarcated as CD25+FOXP3+ (C) and CD25highCD127low (D) population. As shown in E, when CD4+CD25+FOXP3+ (Tregs) population was overlaid on D, the Treg population (dark black dots) corresponds to the CD25highCD127low population indicating that FOXP-3 is expressed by CD25highCD127low population. Furthermore, the frequencies of CD4+CD25+FOXP3+ and CD4+CD25highCD127low population obtained, through concurrent staining, from 9 seronegative individuals were compared and showed 97.6% correspondence between both these populations (F). (TIF 1132 kb)
Figure S2. Bootstrapping exercise to address the effect of unequal sample size on statistical tests. A bootstrapping analysis was carried out to address the difference in numbers of recruited individuals. As the biggest disparity in numbers was between the seronegative (N = 33) and ART naïve HIV-2 group (N = 16), data from 16 individuals of the seronegative group, selected by randomisation (using MS Excel) 10 times, was used for comparison of subset frequencies independently. Following this analysis, where numbers of individuals in comparator groups were similar, we continued to observe a significant increase in the frequency of the Tregs (CD25highCD127low) and effector memory (CD127-CD25-) subset as well as a decline in the fraction of naive/central memory (CD127 + CD25low/−) T cell subset in HIV infected individuals as compared to seronegative controls (bootstrap 1 to 10). Statistical significance was evaluated by unpaired t test; *, p < 0.05; **, p < 0.01; and ***, p < 0.001. (TIF 7989 kb)
Figure S3. Age match analysis to address effect of age on statistical analysis. Age and number of seronegative individuals were matched with HIV infected individuals by selecting the16 oldest individuals from seronegative group and their data was used for the comparison of subsets. The median age of the seronegative individuals was now 40 years and was very similar to the median age of HIV infected individuals as shown in A. Following this analysis, the observations with respect to CD4 + T cell subsets remained unchanged (Fig. B, C and D). With respect to data used for analysis of immune activation and cytotoxicity (Figs. 2 and 3) the age of individuals in each group was compared. The median age of all groups was in the range of 35-44 and 37-44 for immune activation (Fig. E) and cytotoxicity analysis (Fig. F) respectively with no significant statistical differences observed. Thus, both seronegative and HIV infected individuals, for these analyses, seem to be age matched appropriately. (TIF 6315 kb)
Figure S4. Gating strategy for activation marker and granzyme-B. Gating strategy for activation marker: Cells were gated based on characteristic light scatter properties FSC against SSC, followed by gating on CD4+ T cells and CD8+T cells. Thereafter based on expression of HLADR and CD38, CD4+T (Fig. A) and CD8+T (Fig. B) cells were further demarcated as HLADR+CD38+ population. The HLADR+CD38+ population was reported as the activated population. The FMO control was used for gating positive population of CD38. Gating strategy for granzyme-B: The lymphocyte population was gated, followed by gating on CD4+ T cells and CD8+T cells. Thereafter based on expression of granzyme-B, CD4+T (Fig. C) and CD8+T (Fig. D) cells were further analysed for granzyme-B positivity (compared to FMO control) and this population was reported as cytotoxic T cells. At least 1, 00,000 events in the lymphocytes gate were acquired for granzyme-B detection. (TIF 2843 kb)
Salwe, S., Singh, A., Padwal, V. et al. Immune signatures for HIV-1 and HIV-2 induced CD4+T cell dysregulation in an Indian cohort. BMC Infect Dis 19, 135 (2019). https://doi.org/10.1186/s12879-019-3743-7
HIV-2
CD4+T-cell subsets
Immune activation
Immune dysregulation
Longitudinal analysis


text:
Presstek 34DI Digital Offset Press Delivers Exceptional Results for The Printed Word
Presstek, Inc. (NASDAQ: PRST), a leading supplier of digital offset printing solutions to the printing and communications industries, today announced that UK company The Printed Word has installed a Presstek 34DI digital offset press.
Established more than 20 years ago, the Billingshurst company's business model is gradually shifting from its litho roots to an all-digital operation. The ability of the Presstek 34DI to produce affordable high quality printing in short runs is the primary reason The Printed Word added the press to its production platform, explains David Pryer, who runs the firm with his brother Rob. "We were being hard hit by the shift to shorter runs, particularly with runs of 5,000 or less, and we needed to respond to that," he says.
The Printed Word replaced an A1 conventional sheetfed litho press with the 34DI. A B2 press handles the decreasing number of long runs, while a number of colour and black & white digital toner-based presses are used for ultra-short runs and variable data.
"We had nothing perfectly suited to handling medium runs of high quality print," states Pryer, who has been closely monitoring the progress of Presstek DI presses. "We looked at a DI press several years ago. It wasn't the right time for us then, but I kept a close eye on the market. When our conventional litho and toner-based digital presses could no longer handle a significant volume of our mid-range work, we decided to make the investment."
Since installation, the Presstek 34DI has boosted The Printed Word's production capacity and enhanced workflow flexibility. Pryer comments, "We recently moved premises, and that gave us the chance to re-assess our printing capabilities. In that assessment, we found the Presstek 34DI to be the best choice for us because of its fast job-to-job turnover, high quality of print, and the fact that it can be run by one operator. Its automation and waterless printing help make set up much simpler with less for the operator to worry about. The quality far exceeds our expectations, and is better than anyone else can produce. We also like the fact that it has a very small physical footprint."
The Printed Word has recently added web-to-print to its array of services, and the 34DI has played a key role in ensuring that the volume of online print generated can be handled quickly and smoothly. "It has been great," says Pryer. "We can quickly change over from job to job, and it is up and running in just a few minutes. Being able to quickly turn jobs around with the DI press enables us to accommodate more jobs each day."
The company has been running Presstek's chemistry-free Anthem plates on its litho presses for a number of years using a Presstek Dimension CTP system. That made the addition of the 34DI, which images chemistry-free plates on press, straightforward. Pryer adds: "We knew the chemistry-free process well. Both our Dimension and the DI press produce top quality plates, and we don't have the hassle of disposing of chemicals."
Since the installation of the Presstek 34DI, Pryer has been delighted with the results. "It is a great machine," he says. "It does everything we want it to. We haven't had any problems at all."
The Printed Word specializes in transactional and general commercial print for the financial sector with 15 employees and a £1.5m turnover.
Source: www.presstek.com
Etiquetas: Printing Equipment


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text:
Sync is an onboard computer system
designed to seamlessly connect the automobile, the driver
and the driver's electronic devices.
"There is a transition point for most people
between the office and the home,
and for most people, that is the car."
BILL GATES IS WATCHING YOU!
Microsoft Corp.'s Chairman Bill Gates is unveiling products he hopes
will encourage more widespread adoption of the digital or "connected" home,
where multiple devices can access and share multimedia content
stored on a PC or a central server hub.
How the two Bills got in Sync
by Bryce Hoffman, Detroit News, Jan 8, 2007
DEARBORN -- When Ford Motor Co. and Microsoft Corp. unveiled Sync in Detroit and Las Vegas Sunday, there were sighs of relief under all the applause. After all, there was almost nothing to unveil. With three weeks left before Sync's formal announcement at the North American International Auto Show and the Consumer Electronics Show, there was still no sign of a working prototype of the technology that links cars with cell phones and personal music players. Both companies were readying their brave faces for productless press conferences. But when Bill Gates is personally overseeing a project, the pieces have a way of falling into place.
"Officially, I am the executive sponsor for all of Microsoft's work with Ford, which means I help determine partnership ideas and ultimately approve any projects either company proposes," Microsoft's chairman told The Detroit News in an e-mail exchange Saturday. That this project has come together so quickly is a testament to the support Sync has from the highest levels of both companies. Think of it as the tale of two Bills.
Ford Executive Chairman Bill Ford Jr. has enjoyed a close working relationship with Gates, a man whose very name is synonymous with computer technology. "Bill Ford Jr. is someone I respect as having a vision for how technology can improve the car experience overall," Gates said.
Both Ford and Microsoft have long sought a project worthy of their iconic names. For more than three years, senior representatives from both companies have met quarterly to discuss the automaker's internal information technology projects, as well as emerging trends in automotive and computer technology. The sessions rotate between Microsoft's headquarters in Redmond, Wash., and Ford's headquarters in Dearborn. Some are attended by Bill Gates or Bill Ford. Though these meetings have spawned intriguing ideas, Sync is the first actual production product to come out of these sessions.
Sync is an onboard computer system designed to seamlessly connect the automobile, the driver and the driver's electronic devices. It gives the driver almost complete hands-free control of devices like cellular telephones and MP3 music players. Powered by Windows CE, Sync allows motorists to make calls, listen to text messages and cue up songs without their hands ever leaving the steering wheel. Work on Sync began when both companies realized they were seeking a common solution to two very different problems.
Ford wanted to develop its own technology to counter the OnStar system already being offered by rival General Motors Corp. Microsoft was eager to establish a beachhead in the automotive space as part of its broader campaign to drive its products into every facet of consumer's lives. Microsoft has long wanted to crack the automotive market, according to Martin Thall, who heads the company's automotive division, but not for the obvious reasons. "The automotive opportunity is not a traditional Microsoft-level opportunity," Thall said. "(But) there is a transition point for most people between the office and the home, and for most people, that is the car." Making sure its products work better in their owners' cars than products from other manufacturers would give Microsoft a competitive edge. And the only way to guarantee that they do is to control the underlying technology that brings them together. "We feel that's a strategic advantage," Thall said.
In 2003, Microsoft began developing the technology and looking for partners in each of the major automobile markets to put it on the road, partnering with Fiat SpA to create Blue & Me -- a system similar to Sync that debuted at last year's Geneva motor show. Next came North America. Thall said GM was the logical choice because of its size, but Microsoft quickly realized that GM was committed to its own OnStar technology. So, Microsoft approached Ford. In April of 2005, Bill Gates traveled to Dearborn to present a $1 million donation to The Henry Ford Museum. After the formal ceremony, he and Bill Ford met to discuss ways their two companies might be able to collaborate on something like Sync. When the representatives from Ford and Microsoft held their next digital lifestyle council, Sync was on the agenda.
Talks on the project began in the summer of 2005. By November, both companies were on the same page and formal contract negotiations began in December of that year, when a delegation from Dearborn led by Ford product development chief Derrick Kuzak and Ford board member Robert Rubin, who has since resigned, traveled to Redmond to meet with Gates and Thall. "We spent a solid day brainstorming on how we would do this," Thall said, adding that Ford outlined its own vision for how the technology could be incorporated into its cars and trucks. "Both companies left that meeting very committed to moving forward with this project." In April of 2006, a final contract was signed by the two companies and Continental Corp., which would supply the hardware. It called for the first production units to be delivered just a year later.
Work on Sync began before the contract was even signed. In January, 80 of the key Ford employees responsible for the project gathered for a five-day meeting at Ford's Conference and Event Center in Dearborn. Mark Fields, president of Ford's Americas group, was on hand to underscore that sense of urgency. He told employees that the hopes of the company were riding on this project. "The original roll-out of the project extended two or three years. We challenged the team to compress that," Fields told The News in an interview Friday. "We said, 'The world's changing fast, and this is a competitive advantage for us. How do we get it across our vehicle lineup as soon as possible?' They delivered."
For its part, Microsoft devoted 90 percent of its automotive division's resources to the project, and Bill Gates personally guaranteed delivery by the target date. "Every time we go out there, he wants to see us, and meet with us on this and get a status update," VanDagens said. "He wants to see this succeed. Both companies have gone through the normal posturing, and we've had negotiation stops and starts. But we'll say, 'Bill Ford really wants to see this happen.' And they'll say, 'Bill Gates really wants to see this happen.' And, with that kind of support, the teams tend to quickly work out these issues. It's been a tremendous catalyst."
Yet, even with this sort of high-level support in both companies, it has been a real challenge to keep Sync on its accelerated development track. Ford's system was supposed to use the same processors as the already proven Fiat system. By May, however, it had become clear that this hardware was not powerful enough to run Sync, particularly its advanced voice-controlled interface. Four weeks later, engineers had created a new unit with faster, more powerful chips. But on Dec. 21, working prototypes finally arrived from Redmond. It was not fully functional, but it was close enough to wow the journalists assembled in Detroit and Las Vegas. "Microsoft is America's technology leader, and Ford is working hard to be America's car company," Bill Ford told The News Sunday. "There's a shared sense of opportunity between the two companies because we know our technology is going to change people's lives for the better."
Gates expands Microsoft's digital home plan
Elizabeth Montalbano, IDG News, Jan 8, 2007
Microsoft Corp.'s Chairman Bill Gates is kicking off the International Consumer Electronics Show (CES) Sunday by unveiling products Microsoft hopes will encourage more widespread adoption of the digital or "connected" home. Microsoft has been promoting the idea of a connected home, where multiple devices can access and share multimedia content stored on a PC or a central server hub, for some time, but so far only the most savvy or wealthy technology enthusiasts have realized even a piece of that vision. But Gates and Robbie Bach, the president of Microsoft's Entertainment and Devices Division, who also is scheduled to appear Sunday, aim to show how Microsoft can help more people get access to the technology.
Gates is announcing that service providers such as AT & T Corp. that offer IPTV (Internet Protocol Television) using Microsoft's software will begin offering this year the Xbox 360 console in lieu of a set-top box. Service providers still will give users the option of the typical set-top box for IPTV or an Xbox 360, said Microsoft spokesman Eric Hollreiser. Still, the announcement sets up a scenario where a home user can watch television and surf the Internet through their Xbox 360 console, which also is an IP-connected device. The move shows Microsoft upping the ante to provide not just software, but also hardware for the digital home, which could eventually put it in competition with its own hardware partners.
As expected, Windows Vista, which will have its widespread consumer release on Jan. 30, is also a major focus of Gates' speech. He is expected to unveil new deals that will deliver more media content through Vista's Media Center capabilities. Windows Media Center, which used to be its own OS but is now part of Vista, allows users to serve up content stored on their PC on televisions, or use their PC or another device to set content for their TVs. Deals with NASCAR, Fox Sports, Nickelodeon, Showtime and Bongo will deliver specialized content through Windows Media Center. All of the content except Bongo's will be free; users will have to pay a subscription for Bongo content. Microsoft also will allow users to submit video content they have created to Media center by integrating it with Soapbox, Microsoft's YouTube-like user-generated video upload service.
The debut of Windows Home Server, a product Microsoft has mentioned before under the code-name "Quattro," is also aimed at helping consumers establish a more connected home. Windows Home Server will not be sold directly to consumers, but will be used by OEMs such as Hewlett-Packard Co. as the basis for new hardware that consumers can put in their homes to connect their Windows Vista computers, Hollreiser said. Users with a broadband connection and more than one computer or device that has an Internet connection can access data stored on Windows Home Server. It also will provide data security and automatically back up data every night. Windows Home Server is aimed at providing a centralized server hub for multimedia files in the digital home, Hollreiser said. It will come in both Windows Vista and Windows XP versions, and pricing and specifications will vary according to the manufacturer. The first hardware using Windows Home Server software should be available sometime this year.
Gates is also showing off some new hardware from Microsoft partners that use new Vista features. HP's TouchSmart PC, for instance, will take advantage of touchscreen capabilities in Vista, while the Toshiba Portege R400 laptop will include a display screen on the top of the laptop that uses Vista's Sideshow technology to show users content such as their Outlook schedule without having to open or turn on their PC. Gates also will show off a new ultramobile PC from Medion, as well as a new Vaio by Sony Corp. optimized for Media Center.
Gates Wants Connected Cars & Homes, Detroit News, Jan 2007
GATES PLANTING STERILE SEEDS
Gates says big changes in store for Internet in next decade ($280 million to build a research and development center in China's capital Beijing, and will double the number of its full-time research staff in China to 3,000 in three to five years). AP/Breibart, May 6, 2008
FORD-LINDBERGH-ORWELL-HITLER (reader was taken aback by putting Lindbergh, Ford and Orwell together in the same sentence
Reader says my article about Ford, Lindbergh, Orwell & Hitler is ignorant, bigoted & anti-Semitic
Bill Gates hails 'digital decade'. BBC, Jan 8, 2007
In a speech with few concrete announcements, Mr Gates outlined how his firm's latest operating system Vista would be the tool to connect people. "People want to do things with their content across multiple platforms," he said. He said the hardware and content had been put in place "and the key thing missing is the connections"....Microsoft's work in developing IPTV (internet protocol TV), which allows programmes to be delivered live or on demand over an internet connection, would soon come to Xbox 360 games consoles....Mr Gates also confirmed that next year would see his last keynote speech at CES as he steps down as head of Microsoft. He said he had offered to come back the following year but that he might "talk more about infectious diseases than software", referring to his philanthropic work.
Dark cloud over Gates good works ("Gates mistakes"; "healing & stealing"). LATimes, Jan 7, 2007
The Gates Foundation has poured $218 million into polio and measles immunization and research worldwide, including in the Niger Delta. At the same time that the foundation is funding inoculations to protect health, The Times found, it has invested $423 million in Eni, Royal Dutch Shell, Exxon Mobil Corp., Chevron Corp. and Total of France — the companies responsible for most of the flares blanketing the delta with pollution, beyond anything permitted in the United States or Europe. Indeed, local leaders blame oil development for fostering some of the very afflictions that the foundation combats....
Toyota lending Nanny State a hand (sensors in steering wheel, cameras check your pupils). NationalPost, Jan 8, 2007. Go to NANNY STATE IN DRIVER'S SEAT
'Xbox is a PC' says Bill Gates (a strategy to get into living room). CVG, Jan 8, 2007. Go to BILL GATES BEHIND TELESCREEN
1.Winston's Diary and 2.Big Brother and 3.Surveillance and 20.Thought Police


Landlords must be careful who they allow to rent their property. K.V.G. Properties rented real property to criminals who created a marijuana growing facility in the property and, as part of the work to grow the marijuana, did severe damage to the real property.
In K.V.G. Properties, Inc. v. Westfield Insurance Company, No. 17-2421, United States Court Of Appeals For The Sixth Circuit (August 21, 2018) the Sixth Circuit was faced with an insurance-coverage dispute because some of KVG's commercial tenants got caught growing marijuana in their rental units. Unfortunately for their landlord, the tenants caused substantial damage to the premises before the police caught up with them.
KVG, a commercial landlord, leased several pieces of real property to a group of commercial tenants. KVG authorized the tenants to use the property for general office or light industrial business. On October 29, 2015, the U.S. Drug Enforcement Agency raided the premises and caught the tenants growing lots of marijuana. KVG speedily evicted the tenants, but the damage had already been done: To accommodate their "business," the tenants removed walls, cut holes in the roof, altered ductwork, and severely damaged the HVAC systems. The total cost of repair appears to be around $500,000.
KVG subsequently filed a claim with Westfield for insurance coverage. Westfield denied the claim, finding that several exclusions applied. The district court found that the loss was excluded by the policy and granted Westfield's motion for summary judgment.
The Michigan courts engage in a two-step analysis when determining coverage under an insurance policy: (1) whether the general insuring agreements cover the loss and, if so, (2) whether an exclusion negates coverage.
The insuring agreements are written broadly to cover all "Risks of Direct Physical Loss." Indeed, one would struggle to think of damage not covered by this language, and this is not a case that tests its boundaries. It is abundantly clear that the property suffered physical damage, necessarily caused by some risk (or risks) of direct physical loss. The harder question is whether the risks here are not covered because an exclusion takes them off the table.
Westfield argues that KVG's tenants conduct was criminal under either state or federal law and that these acts were the main cause of KVG's loss. Coverage under a policy is lost if any exclusion within the policy applies to an insured's particular claims.
Under this exception, the core issue is whether the tenants committed a "criminal act" within the meaning of the policy. Cultivating marijuana is a crime under federal law but it is protected by Michigan law under certain conditions.
Exercising the Michigan courts' common-law power to interpret public initiatives, a federal court would hesitate before reading a Michigan insurance policy to bar coverage for a "criminal act" when Michigan law confers criminal and civil immunity for the conduct at issue. However, no reasonable jury could find that KVG's tenants complied with Michigan law.
Ultimately, the insurer bears the burden of proving facts showing that an exclusion applies. Here, the record contains evidence that KVG itself claimed, in Michigan court, that its tenants violated the law. In its eviction pleadings against each tenant, KVG repeatedly claimed that the "[t]enant illegally grew marijuana" in the unit and stated that the "[i]llegal growing of marijuana" was a "continuing health hazard." These pleadings were signed by KVG's lawyer, who sought and obtained immediate possession of the premises under Michigan's summary eviction statute.
Neither party disputes that federal agents raided the premises as part of a criminal investigation. At the time of the raids, federal officials were operating under guidance from the Deputy Attorney General stating that they should not prioritize individuals whose actions are in clear and unambiguous compliance with existing state laws providing for the medical use of marijuana.
The facts of the raid and arrests of the tenants were sufficient to meet Westfield's prima facie case of proving a criminal act by the preponderance of the evidence.
The policy says "criminal act," not "crime" or "criminal conviction." A fugitive from justice may properly be deemed a criminal by the person he harms, even though the State cannot prove it beyond a reasonable doubt. The Michigan Supreme Court would not read an onerous "conviction requirement" into a standard commercial insurance contract. The Sixth Circuit declined to do so in the complete absence of state authority providing otherwise.
The policy's insuring agreements cover the damage here. Westfield, however, has proven that the Dishonest or Criminal Acts Exclusion applies. Therefore, the district court did not err in granting Westfield's motion for summary judgment.
The criminal acts exclusion, clearly and unambiguously, excluded coverage for losses due to the acts of a person or persons to whom the insured entrusted the property. In this case the insured entrusted the property to its tenants who damaged the property as part of their illegal marijuana growing operation. The lack of coverage was obvious and criminal.


text:
Nadine Labaki's Capernaum
The Daily — May 19, 2018
Lebanese actor-filmmaker Nadine Labaki moved up to the Cannes competition lineup this year with Capernaum, following her 2007 directorial debut, Caramel, which premiered in the Directors' Fortnight, and her 2011 Un Certain Regard selection Where Do We Go Now? Variety's Jay Weissberg argues that the new film "represents a major leap forward in all departments. Proving herself an astonishingly accomplished director of non-professional performers as well as a measured storyteller, Labaki draws attention to the plight of children in Beirut's slums . . . While this is unquestionably an issue film, it tackles its subject with intelligence and heart."
Capernaum is structured around a trial in which a twelve-year-old, already serving time for stabbing someone, is suing his parents for giving him life. Flashbacks reveal a childhood that is indeed wretched. "Danny Boyle's Slumdog Millionaire is an obvious point of comparison," suggests the Telegraph's Robbie Collin. "But the film is closer in texture and spirit to Fernando Meirelles's Brazilian gangster epic City of God, and has a middle hour so crazily ambitious—imagine a neorealist version of Baby's Day Out—I'm not entirely clear how Labaki pulled it off. Watch it win the Palme here on Saturday, and maybe the foreign language Oscar next."
Many critics will grant that Capernaum is grade A awards bait, but that doesn't mean they have to like it. This is "the kind of social-issue sadness pile that confuses nonstop hardship for drama, begging for our tears at every moment," writes the A.V. Club's A. A. Dowd. "Capernaum's neorealist spirit is smothered by its sentimentality and endless string of indignities." Vulture's Emily Yoshida finds Labaki's direction "unnecessarily manipulative," while for IndieWire's David Ehrlich, "for every moment that feels more witnessed than staged . . . , there's another that feels glaringly artificial."
More from Peter Bradshaw (Guardian, 3/5), Martyn Conterio (CineVue, 4/5), Ben Croll (TheWrap), Mónica Delgado (desistfilm), Leslie Felperin (Hollywood Reporter), Lee Marshall (Screen), and Barbara Scharres (RogerEbert.com).
Nicolas Rapold and Eugene Hernandez discuss Capernaum and more films at Cannes on a new episode of the Film Comment Podcast (44'59").
Variety's Elsa Keslassy reports that Sony Pictures Classics has taken North American and Latin American rights.
For news and items of interest throughout the day, every day, follow @CriterionDaily.
More: The Daily
Did You See This?
This week we're reading Greg Tate on MLK/FBI, Ian Christie on the decadence of early British cinema, and Reverse Shot's 2020 top ten.
On Film / The Daily — Jan 15, 2021
Perspectives on Lynne Sachs
Film About a Father Who is the centerpiece of this month's virtual retrospective.
Gothams, Polls, and Lists
Nomadland has triumphed on the first big night of awards season.
Michael Apted's Project of a Lifetime
Apted's lasting legacy will be the Up series, one of the most moving and influential works of nonfiction ever made.


Classic monkstraps with a contemporary feel. Made of soft calfskin in strict brown tones, with a double side buckle, toecaps, and a leather core. Handcrafted with a Blake Rapid production, symbolising sophisticated simplicity.
Double buckle made of soft calf leather, toe, Cuoio Blake construction process.


text:
Aleurocanthus multispinosus es un hemíptero de la familia Aleyrodidae, con una subfamilia: Aleyrodinae. Fue descrita científicamente en 1961 por Dumbleton.

Referencias

multispinosus

redpajama_set_name: RedPajamaWikipedia


Automatically grants the achievement Gundrak if you have not already obtained it.
The entrance to this instance can be found at two locations in Zul'Drak.
I've done this instance twice, and for some reason I can't get the achievement. It never gives me credit for Eck. Do I have to kill him last or something?
Vaincre les boss de Gundrak en mode héroïque.