PLOS ONE: [sortOrder=DATE_NEWEST_FIRST, sort=Date, newest first, q=subject:"Cell enumeration techniques"]PLOShttps://journals.plos.org/plosone/webmaster@plos.orgaccelerating the publication of peer-reviewed sciencehttps://journals.plos.org/plosone/search/feed/atom?sortOrder=DATE_NEWEST_FIRST&unformattedQuery=subject:%22Cell+enumeration+techniques%22&sort=Date,+newest+firstAll PLOS articles are Open Access.https://journals.plos.org/plosone/resource/img/favicon.icohttps://journals.plos.org/plosone/resource/img/favicon.ico2024-03-28T12:30:29ZAccurate and reproducible enumeration of CD4 T cell counts and Hemoglobin levels using a point of care system: Comparison with conventional laboratory based testing systems in a clinical reference laboratory in CameroonBertrand SagniaFabrice Mbakop GhomsiSylvie MoudourouAna GutierezJules TchadjiSamuel Martin SossoAlexis NdjoloVittorio Colizzi10.1371/journal.pone.02977902024-03-20T14:00:00Z2024-03-20T14:00:00Z<p>by Bertrand Sagnia, Fabrice Mbakop Ghomsi, Sylvie Moudourou, Ana Gutierez, Jules Tchadji, Samuel Martin Sosso, Alexis Ndjolo, Vittorio Colizzi</p>
Background <p>Measurements of CD4 T cells and hemoglobin (Hb) are conventionally used to determine the immunological state and disease progression for HIV-infected patients. We obtained a small lightweight point-of-care device, the BD FACSPresto<sup>TM</sup> in order to demonstrate its ability to deliver CD4 and Hb analysis in comparison with two larger clinical machines the BDFACSCanto<sup>TM</sup> analyzer and Sysmex XN 1000 haematology analyzer. The advantages of using the POC device include access to HIV patient data in remote and in resource limited settings.</p> Method <p>The analytical performance of the BD FACSPresto<sup>TM</sup>, compared with the FACSCanto<sup>TM</sup> II flow cytometer and the Sysmex XN 1000 haematology analyzer was evaluated by testing 241 routine clinical specimens collected in EDTA tubes from patients attending the Immunology and Microbiology laboratory of Chantal BIYA International Reference Centre (Yaounde, Cameroon) between January and May 2016.</p> Results <p>The mean in absolute counts and percentage of CD4 T cells was 606 cells/mL and 25% respectively via the FACSPresto<sup>TM</sup>, and 574 cells/mL and 24% respectively via the BD FACSCanto<sup>TM</sup> II. The mean concentration of Hb levels was 11.90 on the Sysmex XN 1000 and 11.45 via the BD FACSPresto<sup>TM</sup>, A high correlation (R<sup>2</sup> = 0.95, P < 0.001) of Hb level measurements was noted between the BD FACSPresto<sup>TM</sup> and Sysmex XN 1000 hematology analyzer. Overall, a Bland-Altman plot of the differences between the two methods showed an excellent agreement for absolute and percentage CD4 counts and hemoglobin measurements between POC and conventional methods evaluated here. Furthermore, the study demonstrated the ease of use of the BD FACSPresto<sup>TM</sup> POC technology in remote areas.</p> Conclusion <p>The BD FACPresto<sup>TM</sup> is a suitable tool for CD4 enumeration in resource-limited settings, specifically providing a deployable, reliable POC testing option. The BD FACSPresto<sup>TM</sup> performed appropriately in comparison to the conventional reference standard technologies. The BD FACSPresto<sup>TM</sup>, system provides accurate, reliable, precise CD4/%CD4/Hb results on venous blood sampling. The data showed good agreement between the BD FACSPresto<sup>TM</sup>, BD FACSCanto<sup>TM</sup> II and Sysmex XN 1000 XN 1000 systems.</p>Intraspecific trait variation modulates the temperature effect on elemental quotas and stoichiometry in marine <i>Synechococcus</i>Renne HarcourtNathan S. GarciaAdam C. Martiny10.1371/journal.pone.02923372024-03-18T14:00:00Z2024-03-18T14:00:00Z<p>by Renne Harcourt, Nathan S. Garcia, Adam C. Martiny</p>
Diverse phytoplankton modulate the coupling between the ocean carbon and nutrient cycles through life-history traits such as cell size, elemental quotas, and ratios. Biodiversity is mostly considered at broad functional levels, but major phytoplankton lineages are themselves highly diverse. As an example, <i>Synechococcus</i> is found in nearly all ocean regions, and we demonstrate contains extensive intraspecific variation. Here, we grew four closely related <i>Synechococcus</i> isolates in serially transferred cultures across a range of temperatures (16–25°C) to quantify for the relative role of intraspecific trait variation vs. environmental change. We report differences in cell size (p<0.01) as a function of strain and clade (p<0.01). The carbon (<i>Q</i><sub>C</sub>), nitrogen (<i>Q</i><sub>N</sub>), and phosphorus (<i>Q</i><sub>P</sub>) cell quotas all increased with cell size. Furthermore, cell size has an inverse relationship to growth rate. Within our experimental design, temperature alone had a weak physiological effect on cell quota and elemental ratios. Instead, we find systemic intraspecific variance of C:N:P, with cell size and N:P having an inverse relationship. Our results suggest a key role for intraspecific life history traits in determining elemental quotas and stoichiometry. Thus, the extensive biodiversity harbored within many lineages may modulate the impact of environmental change on ocean biogeochemical cycles.Using birth-death processes to infer tumor subpopulation structure from live-cell imaging drug screening dataChenyu WuEinar Bjarki GunnarssonEven Moa MyklebustAlvaro Köhn-LuqueDagim Shiferaw TadeleJorrit Martijn EnserinkArnoldo FrigessiJasmine FooKevin Leder10.1371/journal.pcbi.10118882024-03-06T14:00:00Z2024-03-06T14:00:00Z<p>by Chenyu Wu, Einar Bjarki Gunnarsson, Even Moa Myklebust, Alvaro Köhn-Luque, Dagim Shiferaw Tadele, Jorrit Martijn Enserink, Arnoldo Frigessi, Jasmine Foo, Kevin Leder</p>
Tumor heterogeneity is a complex and widely recognized trait that poses significant challenges in developing effective cancer therapies. In particular, many tumors harbor a variety of subpopulations with distinct therapeutic response characteristics. Characterizing this heterogeneity by determining the subpopulation structure within a tumor enables more precise and successful treatment strategies. In our prior work, we developed PhenoPop, a computational framework for unravelling the drug-response subpopulation structure within a tumor from bulk high-throughput drug screening data. However, the deterministic nature of the underlying models driving PhenoPop restricts the model fit and the information it can extract from the data. As an advancement, we propose a stochastic model based on the linear birth-death process to address this limitation. Our model can formulate a dynamic variance along the horizon of the experiment so that the model uses more information from the data to provide a more robust estimation. In addition, the newly proposed model can be readily adapted to situations where the experimental data exhibits a positive time correlation. We test our model on simulated data (<i>in silico</i>) and experimental data (<i>in vitro</i>), which supports our argument about its advantages.Long-term <i>in vitro</i> monitoring of AAV-transduction efficiencies in real-time with Hoechst 33342Xiaonan HuRoland MeisterJan TodeCarsten FrammeHeiko Fuchs10.1371/journal.pone.02981732024-03-01T14:00:00Z2024-03-01T14:00:00Z<p>by Xiaonan Hu, Roland Meister, Jan Tode, Carsten Framme, Heiko Fuchs</p>
Adeno-associated viral transduction allows the introduction of nucleic fragments into cells and is widely used to modulate gene expressions <i>in vitro</i> and <i>in vivo</i>. It enables the study of genetic functions and disease mechanisms and, more recently, serves as a tool for gene repair. To achieve optimal transduction performance for a given cell type, selecting an appropriate serotype and the number of virus particles per cell, also known as the multiplicity of infection, is critical. Fluorescent proteins are one of the common reporter genes to visualize successfully transduced cells and assess transduction efficiencies. Traditional methods of measuring fluorescence-positive cells are endpoint analysis by flow cytometry or manual counting with a fluorescence microscope. However, the flow cytometry analysis does not allow further measurement in a test run, and manual counting by microscopy is time-consuming. Here, we present a method that repeatedly evaluates transduction efficiencies by adding the DNA-stain Hoechst 33342 during the transduction process combined with a microscope or live-cell imager and microplate image analysis software. The method achieves fast, high-throughput, reproducible, and real-time post-transduction analysis and allows for optimizing transduction parameters and screening for a proper approach.Inflammatory markers in world trade center workers with asthma: Associations with post traumatic stress disorderJuan P. WisniveskyNikita AgrawalJyoti AnkamAdam GonzalezAlex FedermanSteven B. MarkowitzJanette M. BirminghamPaula J. Busse10.1371/journal.pone.02976162024-02-13T14:00:00Z2024-02-13T14:00:00Z<p>by Juan P. Wisnivesky, Nikita Agrawal, Jyoti Ankam, Adam Gonzalez, Alex Federman, Steven B. Markowitz, Janette M. Birmingham, Paula J. Busse</p>
Background <p>Post-traumatic stress disorders (PTSD) is associated with worse asthma outcomes in individuals exposed to the World Trade Center (WTC) site.</p> Research question <p>Do WTC workers with coexisting PTSD and asthma have a specific inflammatory pattern that underlies the relationship with increased asthma morbidity?</p> Study design and methods <p>We collected data on a cohort of WTC workers with asthma recruited from the WTC Health Program. Diagnosis of PTSD was ascertained with a Structured Clinical Interview for DSM-5 (Diagnostic and Statistical Manuel of Mental Disorders) and the severity of PTSD symptoms was assessed with the PTSD Checklist 5. We obtained blood and sputum samples to measure cytokines levels in study participants.</p> Results <p>Of the 232 WTC workers with diagnosis of asthma in the study, 75 (32%) had PTSD. PTSD was significantly associated with worse asthma control (p = 0.002) and increased resource utilization (p = 0.0002). There was no significant association (p>0.05) between most blood or sputum cytokines with PTSD diagnosis or PCL-5 scores both in unadjusted and adjusted analyses.</p> Interpretation <p>Our results suggest that PTSD is not associated with blood and sputum inflammatory markers in WTC workers with asthma. These findings suggest that other mechanisms likely explain the association between PTSD and asthma control in WTC exposed individuals.</p>Clinical evaluation of a fully electronic microfluidic white blood cell analyzerJianye SuiZhongtian LinShahriar AzizpourFei ChenSunanda GaurKelly KeeneFarzad SoleimaniTanaya BhowmickZubaid RafiqueMehdi Javanmard10.1371/journal.pone.02963442024-01-18T14:00:00Z2024-01-18T14:00:00Z<p>by Jianye Sui, Zhongtian Lin, Shahriar Azizpour, Fei Chen, Sunanda Gaur, Kelly Keene, Farzad Soleimani, Tanaya Bhowmick, Zubaid Rafique, Mehdi Javanmard</p>
The White Blood Cell (WBC) count is one of the key parameters signaling the health of the immune system. Abnormal WBC counts often signal a systemic insult to the body such as an underlying infection or an adverse side effect to medication. Typically, the blood collected is sent to a central lab for testing, and results come back within hours, which is often inconvenient and may delay time-sensitive diagnosis or treatment. Here, we present the CytoTracker, a fully electronic, microfluidic based instant WBC analyzer with the potential to be used at point-of-care. The CytoTracker is a lightweight, portable, affordable platform capable of quantifying WBCs within minutes using only 50 μl of blood (approximately one drop of blood). In this study, we clinically evaluated the accuracy and performance of CytoTracker in measuring WBC and granulocyte counts. A total of 210 adult patients were recruited in the study. We validated the CytoTracker against a standard benchtop analyzer (Horiba Point of Care Hematology Analyzer, ABX Micros 60). Linear dynamic ranges of 2.5 k/μl– 35 k/μl and 0.6 k/μl– 26 k/μl were achieved for total WBC count and granulocyte count with correlation coefficients of 0.97 and 0.98. In addition, we verified CytoTracker’s capability of identifying abnormal blood counts with above 90% sensitivity and specificity. The promising results of this clinical validation study demonstrate the potential for the use of the CytoTracker as a reliable and accurate point-of-care WBC analyzer.Relationship between vitreous interleukin-6 levels and vitreous particles findings on widefield optical coherence tomography in posterior uveitisMami TomitaMizuki TagamiNorihiko MisawaAtsushi SakaiYusuke HarunaShigeru Honda10.1371/journal.pone.02972012024-01-17T14:00:00Z2024-01-17T14:00:00Z<p>by Mami Tomita, Mizuki Tagami, Norihiko Misawa, Atsushi Sakai, Yusuke Haruna, Shigeru Honda</p>
Purpose <p>To investigate relationship between vitreous interleukin-6 levels and vitreous particles findings on widefield optical coherence tomography in posterior uveitis.</p> Methods <p>This retrospective study examined vitreous inflammatory cells (hyperreflective particles) of posterior uveitis on widefield optical coherence tomography (WOCT). We examined the number of hyperreflective particles (possibility of vitreous inflammatory cells) observed on WOCT and the correlations with interleukin-6 (IL-6) levels. The relationship between vitreous IL-6 levels and image findings from WOCT from 37 eyes (34 patients) with posterior uveitis were analyzed. Mean patient age was 63.4±15.7 years. (Mean± standard deviation) IL-6 concentration in vitreous humor was 79.9±7380.9 pg/mL Uveitis was infectious in 9 cases and non-infectious in 28 cases with multiplex polymerase chain reaction system. We measured the number and size of vitreous cells in the posterior vitreous, defined as the space between the upper vitreous and the internal limiting membrane on WOCT at the macular, upper, and lower regions. Image analysis software was also used for cell counting.</p> Results <p>A strong correlation was seen between human and software counts. Pearson’s correlation coefficient (PCC) was performed to compare categorial variables (on macular +0.866; upper cavity +0.713; lower cavity +0.568; total vitreous cavity +0.834; <i>P<0</i>.<i>001 each</i>). IL-6 levels correlated with both vitreous cell counts and cell counts observed on macular WOCT (human-counted group +0.339, <i>P = 0</i>.<i>04</i>; software-counted group +0.349, <i>P = 0</i>.<i>03</i>). Infectious uveitis showed higher IL-6 levels (<i>P = 0</i>.<i>016</i>) and high cell counts compared with non-infectious uveitis (<i>P = 0</i>.<i>04</i>).</p> Conclusions <p>Vitreous number of hyperreflective particles (cells) findings on WOCTcorrelated well with human and software cell counts. Vitreous cells findings on WOCT also correlated with IL-6 concentrations on macular.</p>mDia formins form hetero-oligomers and cooperatively maintain murine hematopoiesisZhaofeng LiMeng SuXinshu XiePan WangHonghao BiErmin LiKehan RenLili DongZhiyi LvXuezhen MaYijie LiuBaobing ZhaoYuanliang PengJing LiuLu LiuJing YangPeng JiYang Mei10.1371/journal.pgen.10110842023-12-29T14:00:00Z2023-12-29T14:00:00Z<p>by Zhaofeng Li, Meng Su, Xinshu Xie, Pan Wang, Honghao Bi, Ermin Li, Kehan Ren, Lili Dong, Zhiyi Lv, Xuezhen Ma, Yijie Liu, Baobing Zhao, Yuanliang Peng, Jing Liu, Lu Liu, Jing Yang, Peng Ji, Yang Mei</p>
mDia formin proteins regulate the dynamics and organization of the cytoskeleton through their linear actin nucleation and polymerization activities. We previously showed that mDia1 deficiency leads to aberrant innate immune activation and induces myelodysplasia in a mouse model, and mDia2 regulates enucleation and cytokinesis of erythroblasts and the engraftment of hematopoietic stem and progenitor cells (HSPCs). However, whether and how mDia formins interplay and regulate hematopoiesis under physiological and stress conditions remains unknown. Here, we found that both mDia1 and mDia2 are required for HSPC regeneration under stress, such as serial plating, aging, and reconstitution after myeloid ablation. We showed that mDia1 and mDia2 form hetero-oligomers through the interactions between mDia1 GBD-DID and mDia2 DAD domains. Double knockout of mDia1 and mDia2 in hematopoietic cells synergistically impaired the filamentous actin network and serum response factor-involved transcriptional signaling, which led to declined HSPCs, severe anemia, and significant mortality in neonates and newborn mice. Our data demonstrate the potential roles of mDia hetero-oligomerization and their non-rodent functions in the regulation of HSPCs activity and orchestration of hematopoiesis.Automated, high-throughput quantification of EGFP-expressing neutrophils in zebrafish by machine learning and a highly-parallelized microscopeJohn EfromsonGiuliano FerreroAurélien BègueThomas Jedidiah Jenks DomanClay DugoAndi BarkerVeton SaliuPaul ReameyKanghyun KimMark HarfoucheJeffrey A. Yoder10.1371/journal.pone.02957112023-12-07T14:00:00Z2023-12-07T14:00:00Z<p>by John Efromson, Giuliano Ferrero, Aurélien Bègue, Thomas Jedidiah Jenks Doman, Clay Dugo, Andi Barker, Veton Saliu, Paul Reamey, Kanghyun Kim, Mark Harfouche, Jeffrey A. Yoder</p>
Normal development of the immune system is essential for overall health and disease resistance. Bony fish, such as the zebrafish (<i>Danio rerio</i>), possess all the major immune cell lineages as mammals and can be employed to model human host response to immune challenge. Zebrafish neutrophils, for example, are present in the transparent larvae as early as 48 hours post fertilization and have been examined in numerous infection and immunotoxicology reports. One significant advantage of the zebrafish model is the ability to affordably generate high numbers of individual larvae that can be arrayed in multi-well plates for high throughput genetic and chemical exposure screens. However, traditional workflows for imaging individual larvae have been limited to low-throughput studies using traditional microscopes and manual analyses. Using a newly developed, parallelized microscope, the Multi-Camera Array Microscope (MCAM™), we have optimized a rapid, high-resolution algorithmic method to count fluorescently labeled cells in zebrafish larvae <i>in vivo</i>. Using transgenic zebrafish larvae, in which neutrophils express EGFP, we captured 18 gigapixels of images across a full 96-well plate, in 75 seconds, and processed the resulting datastream, counting individual fluorescent neutrophils in all individual larvae in 5 minutes. This automation is facilitated by a machine learning segmentation algorithm that defines the most in-focus view of each larva in each well after which pixel intensity thresholding and blob detection are employed to locate and count fluorescent cells. We validated this method by comparing algorithmic neutrophil counts to manual counts in larvae subjected to changes in neutrophil numbers, demonstrating the utility of this approach for high-throughput genetic and chemical screens where a change in neutrophil number is an endpoint metric. Using the MCAM™ we have been able to, within minutes, acquire both enough data to create an automated algorithm and execute a biological experiment with statistical significance. Finally, we present this open-source software package which allows the user to train and evaluate a custom machine learning segmentation model and use it to localize zebrafish and analyze cell counts within the segmented region of interest. This software can be modified as needed for studies involving other zebrafish cell lineages using different transgenic reporter lines and can also be adapted for studies using other amenable model species.GLI1+ perivascular, renal, progenitor cells: The likely source of spontaneous neoplasia that created the AGMK1-9T7 cell lineAndrew M. Lewis Jr.Gideon FosehWei TuKeith PedenAdovi AkueMark KuKurugaDaniel RotroffGladys LewisIlya MazoSteven R. Bauer10.1371/journal.pone.02934062023-12-07T14:00:00Z2023-12-07T14:00:00Z<p>by Andrew M. Lewis Jr., Gideon Foseh, Wei Tu, Keith Peden, Adovi Akue, Mark KuKuruga, Daniel Rotroff, Gladys Lewis, Ilya Mazo, Steven R. Bauer</p>
The AGMK1-9T7 cell line has been used to study neoplasia in tissue culture. By passage in cell culture, these cells evolved to become tumorigenic and metastatic in immunodeficient mice at passage 40. Of the 20 x 10<sup>6</sup> kidney cells originally plated, less than 2% formed the colonies that evolved to create this cell line. These cells could be the progeny of some type of kidney progenitor cells. To characterize these cells, we documented their renal lineage by their expression of PAX-2 and MIOX, detected by indirect immunofluorescence. These cells assessed by flow-cytometry expressed high levels of CD44, CD73, CD105, Sca-1, and GLI1 across all passages tested; these markers have been reported to be expressed by renal progenitor cells. The expression of GLI1 was confirmed by immunofluorescence and western blot analysis. Cells from passages 13 to 23 possessed the ability to differentiate into adipocytes, osteoblasts, and chondrocytes; after passage 23, their ability to form these cell types was lost. These data indicate that the cells that formed the AGMK1-9T7 cell line were GLI1+ perivascular, kidney, progenitor cells.Automated cell counting for Trypan blue-stained cell cultures using machine learningLouis KuijpersEdo van VeenLeo A. van der PolNynke H. Dekker10.1371/journal.pone.02916252023-11-28T14:00:00Z2023-11-28T14:00:00Z<p>by Louis Kuijpers, Edo van Veen, Leo A. van der Pol, Nynke H. Dekker</p>
Cell counting is a vital practice in the maintenance and manipulation of cell cultures. It is a crucial aspect of assessing cell viability and determining proliferation rates, which are integral to maintaining the health and functionality of a culture. Additionally, it is critical for establishing the time of infection in bioreactors and monitoring cell culture response to targeted infection over time. However, when cell counting is performed manually, the time involved can become substantial, particularly when multiple cultures need to be handled in parallel. Automated cell counters, which enable significant time reduction, are commercially available but remain relatively expensive. Here, we present a machine learning (ML) model based on YOLOv4 that is able to perform cell counts with a high accuracy (>95%) for Trypan blue-stained insect cells. Images of two distinctly different cell lines, <i>Trichoplusia ni</i> (High Five<sup>TM</sup>; Hi5 cells) and <i>Spodoptera frugiperda</i> (Sf9), were used for training, validation, and testing of the model. The ML model yielded <i>F</i>1 scores of 0.97 and 0.96 for alive and dead cells, respectively, which represents a substantially improved performance over that of other cell counters. Furthermore, the ML model is versatile, as an <i>F</i>1 score of 0.96 was also obtained on images of Trypan blue-stained human embryonic kidney (HEK) cells that the model had not been trained on. Our implementation of the ML model comes with a straightforward user interface and can image in batches, which makes it highly suitable for the evaluation of multiple parallel cultures (e.g. in Design of Experiments). Overall, this approach for accurate classification of cells provides a fast, bias-free alternative to manual counting.Investigation of nasal epithelial cells as a surrogate for bronchial epithelial cells in the research of equine asthmaDiane Frances LeeDavid James EverestWilliam CooleyMark Andrew Chambers10.1371/journal.pone.02939562023-11-09T14:00:00Z2023-11-09T14:00:00Z<p>by Diane Frances Lee, David James Everest, William Cooley, Mark Andrew Chambers</p>
Equine asthma, previously known as Recurrent Airway Obstruction (RAO) or Inflammatory Airway Disease (IAD), is an often-debilitating condition that may severely affect both performance and quality of life. Research is hindered by the low sample numbers of subjects recruited to studies, a consequence in part of the invasive nature of the sampling methods of bronchial brushing and biopsy. We present an alternative method of sampling equine airway epithelial cells, the ‘nasal brush method’ (NBM). Obtained by light brushing of the ventral meatus whilst the horse is under standing sedation, these cells express the same markers of differentiation as their deeper counterparts. Grown as 3-D spheroids or as air-liquid interface cultures, nasal epithelial cells are responsive to the inflammatory cytokine interleukin-13. This may be attenuated by modulation of the Notch signalling pathway using the gamma-secretase inhibitor Semagecestat; a previously unreported finding that cements the link between equine and human asthma research and strengthens the case for a One Health approach in researching asthma pathophysiology and therapeutic intervention.Comparative physiological study of sea cucumbers from eastern waters of United StatesEaint Honey Aung WinSinthia MumuNahian FahimKusum ParajuliElliott BlumenthalRebecca PaluAhmed Mustafa10.1371/journal.pone.02934812023-10-30T14:00:00Z2023-10-30T14:00:00Z<p>by Eaint Honey Aung Win, Sinthia Mumu, Nahian Fahim, Kusum Parajuli, Elliott Blumenthal, Rebecca Palu, Ahmed Mustafa</p>
Sea cucumbers, belonging to the phylum Echinodermata, are known to possess valuable bioactive compounds that have medicinal properties. In several countries, such as Korea, China, and Japan, they are cultured in the aquaculture industries for food and medicinal purposes. Research has shown that different species of sea cucumbers each possesses unique medicinal values. As a result, we strive towards finding species with better health resilience in aquaculture system to be cultured for nutritional and medicinal purposes. In this paper, we compared the physiological and immunological parameters of three species of sea cucumbers, <i>Cucumaria frondosa</i> (<i>C</i>. <i>frondosa</i>), <i>Isostychopus badionotus</i> (<i>I</i>. <i>badionotus</i>), and <i>Pentacta pygmaea</i> (<i>P</i>. <i>Pygmaea</i>) from the waters of the eastern United States as they have not been studied extensively. Four different cells of sea cucumbers, phagocytic, red spherule, white spherule, and vibratile cells, that contribute to their immunity were counted. <i>C</i>. <i>frondosa</i> exhibited the highest concentrations of phagocytic cells, white spherule cells, and vibratile cells, compared to the two other species. Due to its high phagocytic cell concentration, the highest phagocytic capacity was seen in <i>C</i>. <i>frondosa</i> although it was not statistically significant. We also observed that <i>C</i>. <i>frondosa</i> had the highest total cell count and the highest concentration of coelomic protein among the three species. Lastly, <i>C</i>. <i>frondosa</i> possessed the highest lysozyme activity. Taken together, we concluded that <i>C</i>. <i>frondosa</i> is the best of the three species compared to be reared in the aquaculture systems for use in the food and biomedicine industries due to its immunological and physiological properties.HIV-1 subtype diversity and immuno-virological outcomes among adolescents failing antiretroviral therapy in Cameroon: A cohort studyWilly Le roi Togna PaboJoseph FokamDebimeh NjumeDésiré TakouMaria-Mercedes SantoroRaymond Babila NyasaCollins ChenwiMarie Laure MpouelGrace BeloumouEzechiel Semengue Ngoufack JagniAlex Durand NkaAude Christelle Ka’eGeorges TetoBeatrice DambayaSandrine DjupsaDavy Hyacinthe Gouissi AnguechiaMolimbou EvaristeCedric KamtaLionel BalaVirginie LamboEdie Gregory Halle-EkaneVittorio ColizziCarlo Federico PernoAlexis NdjoloRoland Ndip Ndip10.1371/journal.pone.02933262023-10-25T14:00:00Z2023-10-25T14:00:00Z<p>by Willy Le roi Togna Pabo, Joseph Fokam, Debimeh Njume, Désiré Takou, Maria-Mercedes Santoro, Raymond Babila Nyasa, Collins Chenwi, Marie Laure Mpouel, Grace Beloumou, Ezechiel Semengue Ngoufack Jagni, Alex Durand Nka, Aude Christelle Ka’e, Georges Teto, Beatrice Dambaya, Sandrine Djupsa, Davy Hyacinthe Gouissi Anguechia, Molimbou Evariste, Cedric Kamta, Lionel Bala, Virginie Lambo, Edie Gregory Halle-Ekane, Vittorio Colizzi, Carlo Federico Perno, Alexis Ndjolo, Roland Ndip Ndip</p>
Objective <p>We sought to evaluate the variability of HIV-1 and its effect on immuno-virological response among adolescents living with perinatally acquired HIV (APHI).</p> Methods <p>A cohort study was conducted from 2018–2020 among 311 APHI receiving antiretroviral therapy (ART) in Cameroon. Sequencing of protease and reverse transcriptase regions was performed for participants experiencing virological failure, VF, (Plasma viral load, PVL ≥ 1000 RNA copies/ml). HIV-1 subtypes were inferred by phylogeny; immuno-virological responses were monitored at 3-time points (T1-T3). Cox regression modeling was used to estimate adjusted hazard ratios (aHRs) of progression to: CD4 < 250, and PVL > 5log<sub>10</sub>, adjusted for acquired drug resistance, gender, ART line, adherence, and duration on treatment; p < 0.05 was considered statistically significant.</p> Results <p>Of the 141 participants in VF enrolled, the male-female ratio was 1:1; mean age was 15 (±3) years; and median [IQR] duration on ART was 51 [46–60] months. In all phases, 17 viral clades were found with a predominant CRF02_AG (58.2%, 59.4%, and 58.3%). From T1-T3 respectively, there was an increasing CD4 count (213 [154–313], 366 [309–469], and 438 [364–569] cells/mm<sup>3</sup>) and decline log<sub>10</sub> PVL (5.23, 4.43, and 4.43), similar across subtypes. Among participants with CRF02_AG infection, duration of treatment was significantly associated with both rates of progression to CD4 < 250, and PVL > 5log<sub>10</sub>, aHR = 0.02 (0.001–0.52), and aHR = 0.05 (0.01–0.47) respectively. Moreover, four potential new HIV-1 recombinants were identified (CRF02_AG/02D, CRF02_AG/02A1F2, D/CRF02_AG, and AF2/CRF02_AG), indicating a wide viral diversity.</p> Conclusion <p>Among APHI in settings like Cameroon, there is a wide genetic diversity of HIV-1, driven by CRF02_AG and with potential novel clades due to ongoing recombination events. Duration of treatment significantly reduces the risk of disease progression.</p>Temporal changes in zooplankton indicators highlight a bottom-up process in the Bay of Marseille (NW Mediterranean Sea)Théo GarciaDaniela BănaruLoïc GuillouxVéronique CornetGérald GregoriFrançois Carlotti10.1371/journal.pone.02925362023-10-23T14:00:00Z2023-10-23T14:00:00Z<p>by Théo Garcia, Daniela Bănaru, Loïc Guilloux, Véronique Cornet, Gérald Gregori, François Carlotti</p>
Sixteen years (2005–2020) of zooplankton monitoring in the Bay of Marseille (N-W Mediterranean Sea) are analyzed in relation to physical, meteorological, climatic and biotic data. Samples were collected every two weeks by a vertical haul (0–55 m) of a 200 μm plankton net. Different indices characterizing the mesozooplankton are compared: biomass dry weight of four size fractions between 200 and 2000 μm; abundances of the whole of the mesozooplankton and of 13 main taxonomic groups defined from plankton imagery; seasonal onset timing of each zooplankton group; and two other types of indices: the first characterized diversity based on abundance data, and the second was derived from zooplankton size spectra shape. The clearest pattern in the environmental compartment was an overall decreasing trend in nutrients, shifts in phytoplankton metrics (i.e. size structure and particulate organic matter), and changes in winter conditions (i.e. increasing temperatures, precipitation and NAO). Interannual patterns in the mesozooplankton community were: (i) a decrease of total abundance (ii) a decrease in biomass for the four size fractions, with an earlier decrease for the 1000–2000 μm size fraction (in 2008); (iii) a reduced dominance of copepods (calanoids and oithonoids) and a concomitant increase in abundance of other taxonomic groups (crustaceans, pteropods, chaetognaths, salps) which induced higher diversity; (iv) a first shift in size spectra towards smaller sizes in 2009, when the 1000–2000 μm size fraction biomass decreased, and a second shift towards larger sizes in 2013 along with increased diversity; and (iv) a later onset in the phenology for some zooplankton variables and earlier onset for salps. Concomitant changes in the phytoplankton compartment, winter environmental conditions, zooplankton community structure (in size and diversity) and zooplankton phenology marked by a shift in 2013 suggest bottom-up control of the pelagic ecosystem.