An ESCRT-III Polymerization Sequence Drives Membrane Deformation and Fission.

The endosomal sorting complex required for transport-III (ESCRT-III) catalyzes membrane fission from within membrane necks, a process that is essential for many cellular functions, from cell division to lysosome degradation and autophagy. How it breaks membranes, though, remains unknown. Here, we characterize a sequential polymerization of ESCRT-III subunits that, driven by a recruitment cascade and by continuous subunit-turnover powered by the ATPase Vps4, induces membrane deformation and fission. During this process, the exchange of Vps24 for Did2 induces a tilt in the polymer-membrane interface, which triggers transition from flat spiral polymers to helical filament to drive the formation of membrane protrusions, and ends with the formation of a highly constricted Did2-Ist1 co-polymer that we show is competent to promote fission when bound on the inside of membrane necks. Overall, our results suggest a mechanism of stepwise changes in ESCRT-III filament structure and mechanical properties via exchange of the filament subunits to catalyze ESCRT-III activity.

A chemical inhibitor of IST1-CHMP1B interaction impairs endosomal recycling and induces noncanonical LC3 lipidation.

The endosomal sorting complex required for transport (ESCRT) machinery constitutes multisubunit protein complexes that play an essential role in membrane remodeling and trafficking. ESCRTs regulate a wide array of cellular processes, including cytokinetic abscission, cargo sorting into multivesicular bodies (MVBs), membrane repair, and autophagy. Given the versatile functionality of ESCRTs, and the intricate organizational structure of the ESCRT machinery, the targeted modulation of distinct ESCRT complexes is considerably challenging. This study presents a pseudonatural product targeting IST1-CHMP1B within the ESCRT-III complexes. The compound specifically disrupts the interaction between IST1 and CHMP1B, thereby inhibiting the formation of IST1-CHMP1B copolymers essential for normal-topology membrane scission events. While the compound has no impact on cytokinesis, MVB sorting, or biogenesis of extracellular vesicles, it rapidly inhibits transferrin receptor recycling in cells, resulting in the accumulation of transferrin in stalled sorting endosomes. Stalled endosomes become decorated by lipidated LC3, suggesting a link between noncanonical LC3 lipidation and inhibition of the IST1-CHMP1B complex.

Molecular characterization of Rft1, an ER membrane protein associated with congenital disorder of glycosylation RFT1-CDG.

The oligosaccharide needed for protein N-glycosylation is assembled on a lipid carrier via a multistep pathway. Synthesis is initiated on the cytoplasmic face of the endoplasmic reticulum (ER) and completed on the luminal side after transbilayer translocation of a heptasaccharide lipid intermediate. More than 30 congenital disorders of glycosylation (CDGs) are associated with this pathway, including RFT1-CDG which results from defects in the membrane protein Rft1. Rft1 is essential for the viability of yeast and mammalian cells and was proposed as the transporter needed to flip the heptasaccharide lipid intermediate across the ER membrane. However, other studies indicated that Rft1 is not required for heptasaccharide lipid flipping in microsomes or unilamellar vesicles reconstituted with ER membrane proteins, nor is it required for the viability of at least one eukaryote. It is therefore not known what essential role Rft1 plays in N-glycosylation. Here, we present a molecular characterization of human Rft1, using yeast cells as a reporter system. We show that it is a multispanning membrane protein located in the ER, with its N and C termini facing the cytoplasm. It is not N-glycosylated. The majority of RFT1-CDG mutations map to highly conserved regions of the protein. We identify key residues that are important for Rft1's ability to support N-glycosylation and cell viability. Our results provide a necessary platform for future work on this enigmatic protein.

Frontiers in plasma proteome profiling platforms: innovations and applications.

Biomarkers play a crucial role in advancing precision medicine by enabling more targeted and individualized approaches to diagnosis and treatment. Various biofluids, including serum, plasma, cerebrospinal fluid (CSF), saliva, tears, pancreatic cyst fluids, and urine, have been identified as rich sources of potential for the early detection of disease biomarkers in conditions such as cancer, cardiovascular diseases, and neurodegenerative disorders. The analysis of plasma and serum in proteomics research encounters challenges due to their high complexity and the wide dynamic range of protein abundance. These factors impede the sensitivity, coverage, and precision of protein detection when employing mass spectrometry, a widely utilized technology in discovery proteomics. Conventional approaches such as Neat Plasma workflow are inefficient in accurately quantifying low-abundant proteins, including those associated with tissue leakage, immune response molecules, interleukins, cytokines, and interferons. Moreover, the manual nature of the workflow poses a significant hurdle in conducting large cohort studies. In this study, our focus is on comparing workflows for plasma proteomic profiling to establish a methodology that is not only sensitive and reproducible but also applicable for large cohort studies in biomarker discovery. Our investigation revealed that the Proteograph XT workflow outperforms other workflows in terms of plasma proteome depth, quantitative accuracy, and reproducibility while offering complete automation of sample preparation. Notably, Proteograph XT demonstrates versatility by applying it to various types of biofluids. Additionally, the proteins quantified widely cover secretory proteins in peripheral blood, and the pathway analysis enriched with relevant components such as interleukins, tissue necrosis factors, chemokines, and B and T cell receptors provides valuable insights. These proteins, often challenging to quantify in complex biological samples, hold potential as early detection markers for various diseases, thereby contributing to the improvement of patient care quality.

Relapse and transformation to myelodysplastic syndrome and acute myeloid leukemia following immunosuppressive therapy for aplastic anemia is more common as compared to allogeneic stem cell transplantation with a negative impact on survival.

We studied the incidence of relapse, transformation to myelodysplastic syndrome/acute myeloid leukemia, and survival in patients with aplastic anemia (AA) surviving more than 1 year after ATG/ALG-based immunosuppressive therapy (IST) between 1985 and 2020. Four-hundred seventy patients (413 adults and 57 children) were studied, and data were compared with 223 patients who underwent matched sibling donor transplant (MSD HSCT). Median follow-up is 50 months (12-359). Relapse occurred in 21.9% at a median time of 33.5 months (5-228) post IST. Twenty-six (5.5%) patients progressed to PNH, while 20 (4.3%) evolved to MDS/AML. Ten-year estimated overall survival (OS) is 80.9 ± 3% and was significantly better in patients without an event (85.1 ± 4%) compared to relapse (74.6% ± 6.2%) or clonal evolution (12.8% ± 11.8%) (p = 0.024). While the severity of AA (p = 0.011) and type of ATG (p = 0.028) used predicted relapse, only age at IST administration influenced clonal evolution (p = 0.018). Among HSCT recipients, relapse rates were 4.9% with no clonal evolution, and the 10-year OS was 94.5 ± 2%. In patients who survived 1 year following IST, outcomes were good except with clonal evolution to MDS/AML. These outcomes, however, were still inferior compared to matched sibling donor HSCT.

Meta-analysis of the results of haploidentical transplantation in the treatment of aplastic anemia.

This meta-analysis was to evaluate the outcome of haploidentical hematopoietic stem cell transplantation (Haplo-HSCT) for aplastic anemia (AA) compared with matched related donor (MRD)-HSCT, matched unrelated donor (MUD)-HSCT, and immunosuppressive therapy (IST). Pubmed, Embase, Cochrane Library, Web of Science, CNKI, WanFang, and VIP databases were searched for relevant studies from inception to 22 June 2022. Relative risk (RR) was used to indicate the effect indicator, with a 95% confidence interval (CI) being applied to express the effect size. A subgroup analysis based on the literature quality (low, fair, and high) was applied. Totally, 25 studies were included in this study, comprising 2252 patients. Our findings demonstrated no difference between Haplo-HSCT and MRD-HSCT in 1-, 2-, and 3-year overall survival (OS), failure-free survival (FFS), and engraftment. However, Haplo-HSCT had higher incidences of II-IV acute graft-versus-host disease (aGVHD), chronic GVHD (cGVHD), and cytomegalovirus infection. There were no differences in 3- and 5-year OS, 3-year FFS, platelet engraftment, graft failure (GF), II-IV grade of aGVHD, and complication between Haplo-HSCT and MUD-HSCT; however, Haplo-HSCT had a lower incidence of cGVHD. Compared with IST, Haplo-HSCT had a higher 3-year FFS and 3- and 6-month response rate. However, there were no differences in 3- and 5-year OS, and 12-month response rate between Haplo-HSCT and IST. This study suggests that Haplo-HSCT may be a realistic therapeutic option for AA, which may provide a reference for decision-making.

Repeated immunosuppressive rabbit antithymocyte globulin therapy for adult patients with relapsed or refractory aplastic anemia.

OBJECTIVES: Immunosuppressive therapy (IST) with antithymocyte globulin (ATG) and cyclosporin A is the standard treatment for aplastic anemia (AA). However, the efficacy of repeated IST with rabbit ATG (rATG) as salvage therapy remains unclear in patients with relapsed or refractory AA. METHODS: We retrospectively evaluated the efficacy and safety of IST2 with rATG (IST2-rATG) in 19 consecutive patients with relapsed or refractory AA who received first-line IST with rATG in two centers between 2009 and 2020. RESULTS: The overall 6-month response rate of the patients was 58%. The response rates were similar between patients with relapsed and refractory AA. The presence of glycophosphatidylinositol-deficient blood cells was associated with a better response to IST2-rATG. Despite retreatment with the same rATG, serum disease and severe allergic reactions were not observed. CONCLUSION: IST2-rATG is effective and safe for the treatment of adult patients with relapsed and refractory AA after receiving first-line IST with rATG.

Comparison of different measuring methods in the assessment of the ISNT rule and its variants in a normal population: A cross-sectional study.

PURPOSE: The aim of this study was to compare five different neuroretinal rim (NRR) measurement methods, based on quadrants and NRR widths, in the assessment of the ISNT (inferior (I) > superior (S) > nasal (N) > temporal (T)) rule, and its variants IST (inferior (I) > superior (S) > temporal (S)) rule, IS (inferior (I) > superior (S)) rule and T (temporal is the thinnest) rule in a normal population. Factors influencing compliance with this rule and its variants were also evaluated. METHODS: Stereoscopic fundus images were analysed through a dichoptic viewing system. Two graders labelled the optic disc and cup, as well as the fovea. Custom-made software automatically determined the limits of the optic disc and cup and examined the ISNT rule and its variants using several NRR measurement methods. RESULTS: Sixty-nine subjects with normal eyes were enrolled. For the various NRR measuring methods, the percentage of eyes following the rules, that is, validity ranges were 0.0%-15.9% for the ISNT rule, 31.9%-59.4% for the IST rule, 46.4%-59.4% for the IS rule and 50.7%-100.0% for the T rule. Significant intra-measurement agreement ranges were IST (κ = 0.50-0.85), IS (κ = 0.68-1.00) and T (κ = 0.24-0.77). Only the IST and IS rules achieved significant inter-measurement agreement (κ = 0.47-1.00). After multivariate and receiver operating characteristic (ROC) curve analyses, the vertical cup position cupy (area under the ROC curve (AUROC) = 0.60-0.96; cut-off = |0.005|) was the most important predictor for virtually all NRR measurement agreements for the ISNT, IST and IS rules. The horizontal cup position (AUROC = 0.50-0.92; cut-off = -0.028 to 0.05) was the most important predictive factor for the majority of the NRR measurement agreements for the T rule. CONCLUSIONS: Only the IST and IS rules are valid for the same normal subjects. The most important factor affecting the validity of the ISNT rule and its variants was the anatomical cup position. NRR measurement agreements based on NRR quadrants exhibited larger validity and better agreement. The IST and IS rules can be combined with the alternative SIT (superior (S) > inferior (I) > temporal (T)) and SI (superior (S) > inferior (I)) rules to detect almost all normal subjects.

Hepatitis-associated aplastic anaemia: surveillance of frequency,clinico-haematological features, and demographic distribution at a tertiary care hospital in Pakistan.

OBJECTIVE: We aim to document the frequency of HAAA cases among AA patients presenting at a tertiary care hospital, and to determine the most common agents (viral/drug induced) and Clinico-haematological features among HAAA patients at a tertiary care hospital. Methods: This study was a retrospective review, conducted at a tertiary care hospital in Karachi, Pakistan. RESULTS: A total of 21 patients were included in the study. Hepatitis among the HAAA patients was viral in 17 cases, while 4 were idiopathic. All the patients acquired aplastic anaemia within 3-12 months of the Hepatitis episode and most presented with bleeding, bruises and petechiae. CONCLUSIONS: This study indicates and proves that presence and prevalence of this disease in the Pakistani population is quite significant. Unlike the rest of the world, HAAA in Pakistan is not entirely of unknown aetiology, most of the cases can be associated with one of the Hepatitis viruses.

In Search of a Universal Method: A Comparative Survey of Bottom-Up Proteomics Sample Preparation Methods.

Robust, efficient, and reproducible protein extraction and sample processing is a key step for bottom-up proteomics analyses. While many sample preparation protocols for mass spectrometry have been described, selecting an appropriate method remains challenging since some protein classes may require specialized solubilization, precipitation, and digestion procedures. Here, we present a comprehensive comparison of the 16 most widely used sample preparation methods, covering in-solution digests, device-based methods, and commercially available kits. We find a remarkably good performance of the majority of the protocols with high reproducibility, little method dependency, and low levels of artifact formation. However, we revealed method-dependent differences in the recovery of specific protein features, which we summarized in a descriptive guide matrix. Our work thereby provides a solid basis for the selection of MS sample preparation strategies for a given proteomics project.

Structure-Activity relationship of 1-(Furan-2ylmethyl)Pyrrolidine-Based Stimulation-2 (ST2) inhibitors for treating graft versus host disease.

An elevated plasma level of soluble ST2 (sST2) is a risk biomarker for graft-versus-host disease (GVHD) and death in patients receiving hematopoietic cell transplantation (HCT). sST2 functions as a trap for IL-33 and amplifies the pro-inflammatory type 1 and 17 response while suppressing the tolerogenic type 2 and regulatory T cells activation during GVHD development. We previously identified small-molecule ST2 inhibitors particularly iST2-1 that reduces plasma sST2 levels and improved survival in two animal models. Here, we reported the structure-activity relationship of the furanylmethylpyrrolidine-based ST2 inhibitors based on iST2-1. Based on the biochemical AlphaLISA assay, we improved the activity of iST2-1 by 6-fold (∼6 µM in IC50 values) in the inhibition of ST2/IL-33 and confirmed the activities of the compounds in a cellular reporter assay. To determine the inhibition of the alloreactivity in vitro, we used the mixed lymphocyte reaction assay to demonstrate that our ST2 inhibitors decreased CD4+ and CD8+ T cells proliferation and increased Treg population. The data presented in this work are critical to the development of ST2 inhibitors in future.

Clinical outcomes of 34 patients with resistance to thyroid hormone beta: a twenty-year experience in Japan.

Resistance to thyroid hormone beta (RTHß) caused by germline mutations in genes encoding thyroid hormone receptor beta (TRß) is a rare disorder. Little information is available regarding the clinical experience of this syndrome in Japan. We retrospectively reviewed the records of 34 patients with RTHß (21 adult females and 13 adult males) with positive TRß mutations identified at our division between 2000 and 2020. Of the 24 patients with available clinical history, 10 (41.7%) received inappropriate treatments such as antithyroid drugs, thyroidectomy, or radioactive iodine. Diagnostic delay and inappropriate management of RTHß are still present in Japan. Every patient except one demonstrated thyroid hormone profiles indicative of syndrome of inappropriate secretion of thyrotropin (SITSH), characterized by a hormonal profile of hyperthyroxinemia with a non-suppressed TSH concentration. Since the most common forms of hyperthyroidism including Graves' disease feature elevated thyroid hormone levels with suppressed TSH concentrations, early diagnosis of SITSH is critical for preventing inappropriate management. One patient positive for anti-thyroglobulin antibody (Tg-Ab) and anti-thyroperoxidase antibody (TPO-Ab) showed remarkably elevated TSH (>200 µIU/mL) despite thyroid hormone concentrations within the reference ranges. At least one thyroid autoantibody (Tg-Ab, TPO-Ab, or thyrotropin receptor antibodies) was identified in 37.9% (11/29) of the patients tested. One patient developed overt Graves' disease nine years after RTHß diagnosis. These findings suggest that RTHß is frequently comorbid with additional autoimmune thyroid disorders. Further research is required to identify the most appropriate treatments for RTHß patients who develop a second thyroid disorder.

Acquired aplastic anemia complicated with anti-glomerular basement membrane disease successfully treated with immunosuppressive therapy: a case report.

BACKGROUND: Aplastic anemia (AA) is a rare but fatal disorder characterized by pancytopenia due to bone marrow hypoplasia. Anti-glomerular basement membrane disease (anti-GBM disease) is an immune complex small-vessel vasculitis that presents as rapidly progressive glomerulonephritis and/or pulmonary hemorrhage. Although both involve autoreactive T cells that are partially triggered by human leukocyte antigen (HLA)-DR15, there have been no reports of their co-existence and the treatment strategy is not well understood. CASE PRESENTATION: A 67-year-old woman presented with fever, malaise, and acute kidney injury with proteinuria and hematuria requiring hemodialysis. She was diagnosed with anti-GBM antibody disease based on high serum anti-GBM antibody titer and crescentic glomerulonephritis on a renal biopsy. Pulse administration of methylprednisolone (MP), oral prednisolone (PSL), and plasmapheresis were performed. Only 2 weeks after the diagnosis of anti-GBM disease, the patient developed pancytopenia requiring frequent blood transfusions. The blood cell count did not recover even 1 month after discontinuing the drugs that could cause pancytopenia. Bone marrow examination showed hypocellularity without abnormal infiltrates or fibrosis, which led to the diagnosis of severe acquired AA. Further HLA phenotyping revealed that she had HLA-DR15. Increased dose of PSL with the secondary MP pulse and the addition of cyclosporine improved pancytopenia. Although she remained dialysis-dependent, anti-GBM disease and pancytopenia did not recur for more than 2 years. CONCLUSIONS: We report the first case of acquired AA complicated with anti-GBM disease in an elderly woman with HLA-DR15, which was successfully treated with immunosuppressive therapy (IST). This report is valuable not only because it shows they may co-occur, but also because it provides a therapeutic option for this complex condition. It was also suggested that pancytopenia in patients with anti-GBM disease recalls serious hematologic diseases including AA that require immediate treatment based on bone marrow examination.

Profiles and health risks (STIs, HCV, HIV) of injecting drug users.

OBJECTIVE: The aim of this article is twofold: to present the sociodemographic profiles of people who inject drugs (PWID) in Togo and to assess the prevalence of health risks (sexually transmitted infections [STIs], the hepatitis C virus, and HIV), the problems linked to drug injection, and the factors characterizing PWIDs. PARTICIPANTS AND METHODS: Using a questionnaire, this cross-sectional descriptive study was conducted on 384 PWIDs in Togo. The questionnaire focused on sociodemographic characteristics, consumption history, and known health problems and risks. Snowball sampling allowed for data collection in all regions of the country. RESULTS: In the sampling, the results revealed prevalence of 17% for STIs and 53% for the hepatitis C virus. The onset of medical problems and STI signs was significantly triggered when the person was female, over 25 years of age, polygamous, not attending school, unemployed, and had been using drugs for more than five years. Moreover, reused injection equipment was shown to be associated with the high STI prevalence. CONCLUSION: Drug injection is dangerous and results in numerous health problems. This study shows that PWID vulnerability of stems from specific characteristics, such as being uneducated, single, unemployed, bereft of parents, and having a low monthly income. Additional research is required to further investigate the health risks associated with drug injection in view of providing PWIDs with comprehensive care.

Validation of an implant stability measurement device using the percussion response: a clinical research study.

BACKGROUND: Several devices have been developed to measure implant-bone stability as an indicator of successful implant treatment; these include Osstell®, which measures the implant stability quotient (ISQ), and the more recent AnyCheck®, which relies on percussion for the implant stability test (IST). These devices make it possible to measure implant stability. However, no studies have compared the performance of AnyCheck® and Osstell® (i.e., IST and ISQ values) in clinical practice. Therefore, this study aimed to determine the correlation between primary and secondary implant stability using the Osstell® and AnyCheck® devices. METHODS: Ten patients (7 women; age [mean ± standard deviation]: 49.1 ± 13.3 years) with partially edentulous jaws who received a total of 15 implants were included. IST (AnyCheck®) and ISQ (Osstell®) values were measured immediately after implantation and at 1, 2, 3, 4, and 6 weeks post-implantation. Each measurement was performed three times, and the average value was used as the result. The correlation between measurements obtained using the two devices was determined using Spearman's rank correlation coefficient. RESULTS: The IST values ranged from 79.1 ± 2.87 to 82.4 ± 2.65. The ISQ values ranged from 76.0 ± 2.8 to 80.2 ± 2.35. Spearman's rank correlation coefficient was r = 0.64 immediately after implantation, r = 0.29 at 1 week, r = 0.68 at 2 weeks, r = 0.53 at 3 weeks, r = 0.68 at 4 weeks, and r = 0.56 at 6 weeks. A positive correlation was found in all cases, except at week 1 when the correlation was weak; the IST and ISQ values decreased the most during the first postoperative week and increased during the second week. The IST values were also slightly higher at all measurement points. CONCLUSION: The ability to assess implant stability without removing the abutment during healing is essential for determining the timing of loading without the risk of bone resorption. The results of this study suggest that AnyCheck® is useful for determining primary and secondary implant stability.

[Living and aging with HIV]. / Vivre et vieillir avec le VIH.

HIV is not exclusive to the young population. Emotional life continues with age, and 20% of new cases of infection in France are among people over 50. This is therefore a public health issue for "seniors".

Anhydrobiosis in yeast: role of cortical endoplasmic reticulum protein Ist2 in Saccharomyces cerevisiae cells during dehydration and subsequent rehydration.

Two Saccharomyces cerevisiae strains, BY4741 and BY4741-derived strain lacking the IST2 gene (ist2Δ), were used to characterise the possible role of cortical endoplasmic reticulum (ER) protein Ist2 upon cell dehydration and subsequent rehydration. For the first time, we show that not only protein components of the plasma membrane (PM), but also at least one ER membrane protein (Ist2) play an important role in the maintenance of the viability of yeast cells during dehydration and subsequent rehydration. The low viability of the mutant strain ist2∆ upon dehydration-rehydration stress was related to the lack of Ist2 protein in the ER. We revealed that the PM of ist2∆ strain is not able to completely restore its molecular organisation during reactivation from the state of anhydrobiosis. As the result, the permeability of the PM remains high regardless of the type of reactivation (rapid or gradual rehydration). We conclude that ER protein Ist2 plays an important role in ensuring the stability of molecular organisation and functionality of the PM during dehydration-rehydration stress. These results indicate an important role of ER-PM interactions during cells transition into the state of anhydrobiosis and the subsequent restoration of their physiological activities.

An Update on the Pathophysiology and Diagnosis of Inappropriate Secretion of Thyroid-Stimulating Hormone.

Inappropriate secretion of thyroid-stimulating hormone (IST), also known as central hyperthyroidism, is a clinical condition characterized by elevated free thyroxine and triiodothyronine concentrations concurrent with detectable thyroid-stimulating hormone (TSH) concentrations. Similarly, the term syndrome of IST (SITSH) is widely used in Japan to refer to a closely related condition; however, unlike that for IST, an elevated serum free triiodothyronine concentration is not a requisite criterion for SITSH diagnosis. IST or SITSH is an important indicator of resistance to thyroid hormone ß (RTHß) caused by germline mutations in genes encoding thyroid hormone receptor ß (TRß) and TSH-secreting pituitary adenoma. Recent evidence has accumulated for several conditions associated with IST, including RTH without mutations in the TRß gene (non-TR-RTH), the phenomenon of hysteresis involving the hypothalamus-pituitary-thyroid axis (HPT-axis), methodological interference, and Cushing's syndrome after surgical resection. However, little information is available on the systematic pathophysiological aspects of IST in previous review articles. This report presents an overview of the recent advances in our understanding of the etiological aspects of IST that are relevant for diagnosis and treatment. Moreover, the report focuses on the potential mechanism of IST caused by hysteresis in the HPT-axis (lagging TSH recovery) in terms of epigenetic regulation.

Urine Proteomics: Evaluation of Different Sample Preparation Workflows for Quantitative, Reproducible, and Improved Depth of Analysis.

The growing field of urinary proteomics shows promise to expand the number of biomarkers for the diagnosis and prognosis of a number of human diseases. With the rapid developments in mass spectrometry methods for proteome quantification, there exists an opportunity for improved sample processing and separation workflows to make important contributions to urine proteomic analyses. Here we evaluate the performance of four sample preparation methods: MStern, PreOmics in-StageTip (iST), suspension-trapping (S-Trap), and conventional urea In-Solution trypsin hydrolysis for nondepleted urine samples. Data-dependent acquisition (DDA) mode on a QExactive HF mass spectrometer was used for single-shot label-free data acquisition. Our results demonstrate a high degree of reproducibility within each workflow. PreOmics iST yields the best digestion efficiency, whereas the S-Trap workflow gives the greatest number of peptide and protein identifications. Using the S-Trap method and starting with ∼0.5 mL, we identify ∼1500 protein groups and ∼17 700 peptides from DDA analysis with a single injection on the mass spectrometer.

HIV/AIDS awareness and testing practices among adolescents in eastern Ethiopia.

OBJECTIVE: To examine HIV/AIDS awareness, HIV testing practices and associated factors among adolescents in two eastern Ethiopian communities. METHODS: Community-based, cross-sectional study among 2010 adolescents aged 10-19 years. Participants were asked about their awareness of HIV/AIDS and HIV testing practices, and whether they had ever been tested for HIV. Regression models were applied to identify the factors of statistical significance at P-value < 0.05. RESULTS: Of 90% were aware of HIV/AIDS, but only a quarter had ever been tested for HIV. Rural adolescents were less aware of HIV than urban adolescents (AOR = 0.16; 95% CI: 0.05, 0.58), and in-school adolescents had more knowledge about HIV/AIDS than that out-of-school adolescents (AOR = 2.79; 95% CI: 1.88, 4.15). Factors associated with lower uptake of HIV testing were male sex (AOR = 0.74; 95% CI; 0.58, 0.91) and being from a rural area (AOR = 0.16; 95% CI: 0.07, 0.36). Factors associated with higher uptake of HIV testing were being in school (AOR = 1.66; 95% CI: 1.16, 2.38), using the Internet (AOR = 1.52; 95% CI: 1.01, 2.28), and ever visiting a health facility (AOR = 1.54; 95% CI: 1.21, 1.96). CONCLUSIONS: Awareness of HIV/AIDS was high, whereas HIV testing was rare. HIV awareness programs for adolescents should target rural and out-of-school adolescents. Programmes to increase HIV testing implemented in these and similar communities should focus on male and rural adolescents.

OBJECTIF: Examiner la sensibilisation au VIH/SIDA, les pratiques de dépistage du VIH et les facteurs associés chez les adolescents de deux communautés dans l'est de l'Ethiopie. MÉTHODES: Etude transversale, à base communautaire auprès de 2.010 adolescents âgés de 10 à 19 ans. Les participants ont été interrogés sur leurs connaissances sur le VIH/SIDA et sur les pratiques de dépistage du VIH, et s'ils avaient déjà subi un test de dépistage du VIH. Des modèles de régression ont été appliqués pour identifier les facteurs ayant une signification statistique à une valeur P < 0,05. RÉSULTATS: 90% des participants étaient au courant du VIH/SIDA, mais seulement un quart avait déjà subi un test de dépistage du VIH. Les adolescents ruraux étaient moins au courant du VIH que les adolescents urbains (AOR = 0,16; IC95%: 0,05-0,58), et les adolescents scolarisés avaient plus de connaissances sur le VIH/SIDA que les adolescents non scolarisés (AOR = 2,79; IC95%: 1,88-4,15). Les facteurs associés à une moindre adoption du test de dépistage du VIH étaient le sexe masculin (AOR = 0,74; IC95%: 0,58-0,91) et provenir d'une zone rurale (AOR = 0,16; IC95%: 0,07-0,36). Les facteurs associés à une plus grande adoption du test de dépistage du VIH étaient le fait d'être scolarisé (AOR = 1,66; IC95%: 1,16-2,38), l'utilisation d'Internet (AOR = 1,52; IC95%: 1,01, 2,28) et avoir déjà visité un établissement de santé (AOR = 1,54; IC95%: 1,21-1,96). CONCLUSIONS: La sensibilisation au VIH/SIDA était élevée alors que le dépistage du VIH était rare. Les programmes de sensibilisation au VIH devraient cibler les adolescents des zones rurales et ceux non scolarisés. Les programmes pour augmenter le dépistage du VIH, mis en œuvre dans ces communautés et dans des communautés similaires, devraient se concentrer sur les adolescents masculins et ceux vivant en milieu rural.