bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of a Cryopreservation Protocol for Type A Spermatogonia
Authors: IZADYAR FARIBORZ , MATTHIJS‐RIJSENBILT J. J. , DEN OUDEN KRISTA , CREEMERS LAURA B. , WOELDERS HENRI , de ROOIJ DIRK G. 
Published: October 2023

ABSTRACTThe aim of this study was to develop a cryopreservation protocol for type A spermatogonia. Testes from 5‐ to 7‐month‐old calves were collected, and type A spermatogonia were isolated using two‐step enzymatic digestion and Percoll separation. Cells were resuspended in minimum essential medium (MEM) supplemented with 1% bovine serum albumin (BSA) in a final concentration of 6 × 106 per mL, and the effects of different cryoprotectants and freezing protocols were tested. Cells frozen/thawed in medium containing 10% fetal calf serum (FCS) and 1.4 M glycerol or dimethyl sulfoxide (DMSO) had a significantly (P < .05) higher percentage of living cells compared to medium with only FCS, whereas DMSO gave a significantly better cell survival rate than glycerol did. An increase in the concentration of FCS in the DMSO‐based medium to 20% had no effect on survival after freezing and thawing. Furthermore, inclusion of 0.07, 0.14, or 0.21 M sucrose in DMSO‐based medium resulted in a significant improvement of cell survival, cell proliferation in culture, and colonization efficiency in recipient testes. A controlled slow‐freezing rate (1 °C/min) resulted in significantly (P < .05) more viable cells than fast (5°C/min) freezing. However, noncontrolled‐rate freezing, with a comparably low cooling rate, gave even better results than the controlled‐rate slow freezing. Cryopreservation in MEM‐based medium containing 10% FCS, 10% DMSO, and 0.07 M sucrose using a non‐controlled‐rate freezing protocol appeared to be a simple and effective way to preserve type A spermatogonia, with a high yield of almost 70% living cells after thawing. Frozen/thawed spermatogonia survived in culture and retained the ability to proliferate as determined by colorimetric and bromodeoxyuridine incorporation assays. To test whether the stem cells among the A spermatogonia retained their ability to colonize the testis of a recipient mouse, bovine spermatogonia were transplanted. This resulted in colonization 2–3 months after transplantation. In conclusion, for the first time, a method specific for cryopreservation of type A spermatogonia, including spermatogonial stem cells was developed, which allows long‐term preservation of these cells without apparent harmful effects to their function.
Environmental Agents and Erectile Dysfunction: A Study in a Consulting Population
Authors: OLIVA ALEJANDRO , GIAMI ALAIN , MULTIGNER LUC 
Published: October 2023

ABSTRACTWe evaluated chemical and physical environmental agents as risk factors for erectile dysfunction among a consulting population. We studied 199 men who sought medical help for erectile disorders between 1996 and 1998 in 3 andrology units in the Litoral Sur region of Argentina. Patients were evaluated by monitoring nocturnal penile tumescence and rigidity, and were classified as having normal (n = 26), irregular (dissociation, short episode or low amplitude, n = 146), or flat erectile pattern (n = 26). Exposure to environmental agents was assessed by a detailed interview, and 4 groups were constituted: nonexposed, pesticide‐exposed, solventexposed, and heat‐exposed. A multivariate polytomous logistic regression model was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for association between quality of nocturnal erections and exposure groups adjusted for confounding factors. Exposure to environmental agents was a risk factor for a flat erectile pattern (OR 7.1, 95% CI 1.5–33.0 for pesticides; OR 12.2, 95% CI 1.2–124.8 for solvents; and OR 1.7, 95% CI 0.3–9.4 for heat). Associations were much weaker for an irregular erectile pattern (OR 1.8, 95% CI 0.5–6.7 for pesticides; OR 2.1, 95% CI 0.3–17.9 for solvents; and OR 1.2, 95% CI 0.4‐4.0 for heat). Our results suggest that environmental agents constitute a risk factor for erectile dysfunction by interfering with erectile ability.
Cytofluorographic Identification of Activated T‐cell Subpopulations in the Semen of Men With Spinal Cord Injuries
Authors: BASU SARMISTHA , LYNNE CHARLES M. , RUIZ PHILLIP , ABALLA TEODORO C. , FERRELL SEAN M. , BRACKETT NANCY L. 
Published: October 2023

ABSTRACTThe semen of most men with spinal cord injury (SCI) contains an abundance of leukocytes. It is not known if this leukocytospermia contributes to the abnormally low sperm motility observed in many of these men. Our study used flow cytometry to identify the leukocyte population in the semen of 12 men with SCI compared to 8 healthy age‐matched control subjects. The results showed that, compared to control subjects, the semen of men with SCI had increased numbers of mature granulocytes and lymphocytes. The largest proportion of the leukocytes consisted of lymphocytes, and immunophenotypic analysis showed that the greater fraction were T cells, many of which coexpressed human leukocyte antigen HLA‐DR and CD25, suggesting they were in an “activated” state. No significant B‐cell population was evident. Our finding of immunologically active leukocytes is a significant step in understanding the relationship of leukocytospermia and decreased sperm motility in the semen of men with SCI.
Effects of Sildenafil on Nocturnal Penile Tumescence and Rigidity in Normal Men: Randomized, Placebo‐Controlled, Cross‐over Study
Authors: ROCHIRA VINCENZO , GRANATA ANTONIO R. M. , BALESTRIERI ANTONIO , MADEO BRUNO , CARANI CESARE 
Published: October 2023

ABSTRACTWe studied the effects of sildenafil on sleep‐related erections in 44 adult healthy men not affected by erectile dysfunction (mean age ± SD: 39.3 ± 10.5 years). No subjects were administered any medication the first night, but all were randomly administered sildenafil 50 mg or placebo the second night and vice versa the third night. Sildenafil and placebo were administered 1 hour before bedtime. The following parameters of sleep‐related erections, after taking sildenafil or placebo, were analyzed: total number of valid erections, total duration of rigidity more than or equal to 70% of a tightening force of 2.8 N applied by the recording device, total duration of increase in penile circumference more than or equal to 30 mm, maximum rigidity, mean of maximum rigidity, and maximum increase of tumescence. Apart from the maximum increase of tumescence, all the parameters analyzed were significantly higher after sildenafil than after placebo administration during the first 4 hours of monitoring in all subjects (n = 44) (study 1). All the parameters were significantly higher after sildenafil than after placebo administration during the whole 8 hours of monitoring in 25 of 44 subjects (study 2A) who slept at least 8 hours. Comparing both the first and the second 4 hours in the 25 of 44 subjects who slept at least 8 hours (study 2B), all the parameters were significantly higher after sildenafil than after placebo administration, apart from maximum rigidity and mean of maximum rigidity during the first 4 hours. Our data suggest that sildenafil, administered at bedtime, is efficacious in improving sleep‐related erections in normal men, indirectly confirming that the nitric oxide pathway is crucial in the physiology of erections during sleep. The effect of sildenafil is prolonged up to 8–9 hours after its administration.
Investigation of the Differences in the Pharmacokinetics of CYP2D6 Substrates, Desipramine, and Dextromethorphan in Healthy African Subjects Carrying the Allelic Variants CYP2D6*17 and CYP2D6*29, When Compared with Normal Metabolizers
Authors: Marasanapalle Venugopal P. , Masimirembwa Collen , Sivasubramanian Rama , Sayyed Sarfaraz , Weinzierl‐Hinum Armin , Mehta Dheeraj , Kapungu Nyasha Nicole , Kanji Comfort , Thelingwani Roslyn , Zack Julia 
Published: October 2023

AbstractThis study investigated the differences in the pharmacokinetics (PK) of dextromethorphan and desipramine in healthy African volunteers to understand the effect of allelic variants of the human cytochrome P450 2D6 (CYP2D6) enzyme, namely the diplotypes of CYP2D6*1/*2 (*1*1, *1*2, *2*2) and the genotypes of CYP2D6*17*17 and CYP2D6*29*29. Overall, 28 adults were included and split into 3 cohorts after genotype screening: CYP2D6*1/*2 (n = 12), CYP2D6*17*17 (n = 12), and CYP2D6*29*29 (n = 4). Each subject received a single oral dose of dextromethorphan 30 mg syrup on day 1 and desipramine 50 mg tablet on day 8. The PK parameters of area under the plasma concentration‐time curve from time of dosing to time of last quantifiable concentration (AUClast), and extrapolated to infinity (AUCinf), and the maximum plasma concentration (Cmax) were determined. For both dextromethorphan and desipramine, AUCinf and Cmax were higher in subjects of the CYP2D6*29*29 and CYP2D6*17*17 cohorts, as compared with subjects in the CYP2D6*1/*2 diplotype cohort and with normal metabolizers from the literature. All PK parameters, including AUCinf, Cmax, and the elimination half‐life, followed a similar trend: CYP2D6*17*17 > CYP2D6*29*29 > CYP2D6*1/*2. The plasma and urinary drug/metabolite exposure ratios of both drugs were higher in subjects of the CYP2D6*17*17 and CYP2D6*29*29 cohorts, when compared with subjects in the CYP2D6*1/*2 diplotype cohort. All adverse events were mild, except in 1 subject with CYP2D6*17*17 who had moderately severe headache with desipramine. These results indicate that subjects with CYP2D6*17*17 and CYP2D6*29*29 genotypes were 5‐10 times slower metabolizers than those with CYP2D6*1/*2 diplotypes. These findings suggest that dose optimization may be required when administering CYP2D6 substrate drugs in African patients. Larger studies can further validate these findings.
Systematic compositional analysis of exosomal extracellular vesicles produced by cells undergoing apoptosis, necroptosis and ferroptosis
Authors: Cappe Benjamin , Vadi Mike , Sack Eliza , Wacheul Ludivine , Verstraeten Bruno , Dufour Sara , Franck Julien , Xie Wei , Impens Francis , Hendrix An , Lafontaine Denis L. J. , Vandenabeele Peter , Riquet Franck B. 
Published: October 2023

AbstractFormation of extracellular vesicles (EVs) has emerged as a novel paradigm in cell‐to‐cell communication in health and disease. EVs are notably produced during cell death but it had remained unclear whether different modalities of regulated cell death (RCD) influence the biogenesis and composition of EVs. To this end, we performed a comparative analysis of steady‐state (ssEVs) and cell death‐associated EVs (cdEVs) following TNF‐induced necroptosis (necEVs), anti‐Fas‐induced apoptosis (apoEVs), and ML162‐induced ferroptosis (ferEVs) using the same cell line. For each RCD condition, we determined the biophysical and biochemical characteristics of the cell death‐associated EVs (cdEVs), the protein cargo, and the presence of methylated ribosomal RNA. We found that the global protein content of all cdEVs was increased compared to steady‐state EVs. Qualitatively, the isolated exosomal ssEVs and cdEVs, contained a largely overlapping protein cargo including some quantitative differences in particular proteins. All cdEVs were enriched for proteins involved in RNA splicing and nuclear export, and showed distinctive rRNA methylation patterns compared to ssEVs. Interestingly, necEVs and apoEVs, but strikingly not ferEVs, showed enrichment of proteins involved in ribosome biogenesis. Altogether, our work documents quantitative and qualitative differences between ssEVs and cdEVs.
Validation of a surgical difficulty grading system in laparoscopic cholecystectomy for acute cholecystitis
Authors: Asai Koji , Ohyama Tetsuji , Watanabe Manabu , Moriyama Hodaka , Kujiraoka Manabu , Watanabe Ryutaro , Shigeta Kenta , Kakizaki Nanako , Hagiwara Osahiko , Saida Yoshihisa 
Published: October 2023

AbstractBackgroundThe surgical difficulty of laparoscopic cholecystectomy (LC) for acute cholecystitis varies from case to case, and appropriate intraoperative evaluation would help prevent bile duct injury (BDI).MethodsWe analyzed 178 patients who underwent LC for acute cholecystitis. Expert surgeons and trainees individually evaluated the surgical difficulty. The inter‐rater agreement was analyzed using Conger's κ and Gwet's agreement coefficient (AC). Furthermore, we analyzed the predictive surgical difficulty item for performing subtotal cholecystectomy (STC).ResultsRegarding the inter‐rater agreement between expert surgeons and trainees, 15 of the 17 surgical difficulty items had a Gwet's AC of 0.5 or higher, indicating “moderate” agreement or higher. Furthermore, the highest and total surgical difficulty scores were deemed “substantial” agreement. Scarring and dense fibrotic changes around the Calot's triangle area with easy bleeding with/without necrotic changes were predictive of whether STC should be performed.ConclusionsThis surgical difficulty grading system is expected to be a tool that can be used by any surgeon with LC experience. STC should be performed to prevent BDI according to the changes around the Calot's triangle area.
Effects of transcranial direct current stimulation on the glutamatergic pathway in the male rat hippocampus after experimental focal cerebral ischemia
Authors: Akçay Guven , Aslan Mutay , Kipmen Korgun Dijle , Çeker Tugçe , Akan Ezgi , Derin Narin 
Published: October 2023

AbstractThis study aimed to assess the focal cerebral ischemia‐induced changes in learning and memory together with glutamatergic pathway in rats and the effects of treatment of the animals with transcranial Direct Current Stimulation (tDCS). One hundred male rats were divided into five groups as sham, tDCS, Ischemia/Reperfusion (IR), IR + tDCS, and IR + E‐tDCS groups. Learning, memory, and locomotor activity functions were evaluated by behavioral experiments in rats. Glutamate and glutamine levels, alpha‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazole‐propionate receptor (AMPAR1), N‐Methyl‐D‐Aspartate receptors (NMDAR1 and NMDAR2A), vesicular glutamate transporter‐1 (VGLUT‐1), and excitatory amino acid transporters (EAAT1‐3) mRNA expressions in hippocampus tissues were measured. Ischemic areas were analyzed by TTC staining. The increase was observed in IR + tDCS, and IR + E‐tDCS groups compared to the IR group while a significant decrease was observed in IR group compared to the sham in the locomotor activity, learning, and memory tests. While glutamate and glutamine levels, AMPAR1, NMDAR1, NMDAR2A, VGLUT1, and EAAT1 mRNA expressions were significantly higher in IR group compared to the sham group, it was found to be significantly lower in IR + tDCS and IR + E‐tDCS groups compared to the IR group. EAAT2 and EAAT3 mRNA expressions were significantly higher in IR + tDCS and IR + E‐tDCS groups compared to the IR group. Ischemic areas were significantly decreased in IR + tDCS and IR + E‐tDCS groups compared to the IR group. Current results suggest that tDCS application after ischemia improves learning and memory disorders and these effects of tDCS may be provided through transporters that regulate glutamate levels.
Estimation of wheat crop production using multispectral information fusion
Authors: Jitendra  , Pant Triloki 
Published: October 2023

AbstractBACKGROUNDThe present work estimates the area and corresponding wheat crop production in the study area, which comprises the Etah region of Uttar Pradesh, India. For this purpose, multispectral images of multiple sensors, namely Sentinel‐2, Landsat‐8 and Landsat‐9 during the preharvest period, i.e. March for the years 2021 and 2022, were used. A multispectral information fusion approach was proposed, involving image classification as well as vegetation index‐based information extraction. For imposing information fusion, appropriate image bands were identified with the help of separability analysis followed by land cover classification for wheat crop class extraction. Support vector machine (SVM), artificial neural network (ANN) and maximum likelihood (ML) were used for classification, whereas normalized difference vegetation index (NDVI) and fractional vegetation cover (FVC) were used for index‐based crop area extraction.RESULTSA maximum accuracy of 98.34% was achieved for Sentinel‐2 data using ANN, whereas a minimum accuracy of 80.21% was achieved for Landsat‐9 using the ML classifier. The estimated area for Sentinel‐2 data for the year 2021 was 260 540 ha using ANN and 203 240 ha using ML, which is close to the reference data, i.e. 238 600 ha. SVM also showed good performance and calculated least error in estimated crop area for the year 2022 on Sentinel‐2 data. It calculated 8 408 490 tons of wheat for the same year.CONCLUSIONThe proposed method utilizes a single image per year for extraction of information supported by the ground truth data, which makes it a novel approach to information extraction for crop production monitoring. © 2023 Society of Chemical Industry.
Wharton's Jelly mesenchymal stromal cell‐derived extracellular vesicles promote nucleus pulposus cell anabolism in an in vitro 3D alginate‐bead culture model
Authors: Tilotta Veronica , Vadalà Gianluca , Ambrosio Luca , Di Giacomo Giuseppina , Cicione Claudia , Russo Fabrizio , Darinskas Adas , Papalia Rocco , Denaro Vincenzo 
Published: October 2023

AbstractBackgroundIntradiscal transplantation of mesenchymal stromal cells (MSCs) has emerged as a promising therapy for intervertebral disc degeneration (IDD). However, the hostile microenvironment of the intervertebral disc (IVD) may compromise the survival of implanted cells. Interestingly, studies reported that paracrine factors, such as extracellular vesicles (EVs) released by MSCs, may regenerate the IVD. The aim of this study was to investigate the therapeutic effects of Wharton's Jelly MSC (WJ‐MSC)‐derived EVs on human nucleus pulposus cells (hNPCs) using an in vitro 3D alginate‐bead culture model.MethodsAfter EV isolation and characterization, hNPCs isolated from surgical specimens were encapsulated in alginate beads and treated with 10, 50, and 100 μg/mL WJ‐MSC‐EVs. Cell proliferation and viability were assessed by flow cytometry and live/dead staining. Nitrite and glycosaminoglycan (GAG) content was evaluated through Griess and 1,9‐dimethylmethylene blue assays. hNPCs in alginate beads were paraffin‐embedded and stained for histological analysis (hematoxylin–eosin and Alcian blue) to assess extracellular matrix (ECM) composition. Gene expression levels of catabolic (MMP1, MMP13, ADAMTS5, IL6, NOS2), anabolic (ACAN), and hNPC marker (SOX9, KRT19) genes were analyzed through qPCR. Collagen type I and type II content was assessed with Western blot analysis.ResultsTreatment with WJ‐MSC‐EVs resulted in an increase in cell content and a decrease in cell death in degenerated hNPCs. Nitrite production was drastically reduced by EV treatment compared to the control. Furthermore, proteoglycan content was enhanced and confirmed by Alcian blue histological staining. EV stimulation attenuated ECM degradation and inflammation by suppressing catabolic and inflammatory gene expression levels. Additionally, NPC phenotypic marker genes were also maintained by the EV treatment.ConclusionsWJ‐MSC‐derived EVs ameliorated hNPC growth and viability, and attenuated ECM degradation and oxidative stress, offering new opportunities for IVD regeneration as an attractive alternative strategy to cell therapy, which may be jeopardized by the harsh microenvironment of the IVD.
Preliminary Experience with 3‐Dimensional Exoscope‐Assisted Laryngoplasty
Authors: Sanuki Tetsuji , Takemoto Naoki , Sumiya Shogo , Iwasaki Shinichi 
Published: October 2023

ObjectiveLaryngoplasty requires the manipulation of the vocal folds, which are not visible during the operation. The widespread use of this technique is limited by the need for adequate knowledge of anatomy, the small surgical field, and the high level of skill required for the procedure. An exoscope has been developed to provide a stereoscopic view similar to that of a microscope while using the same compact endoscopic tool. This study aimed to determine whether the three‐dimensional (3D) exoscopic surgical technique could be applied to laryngoplasty and explore its possibility to ultimately replace the current approach.MethodsThis was a retrospective case series analysis, which included 28 patients with hoarseness who underwent surgery with (Exoscope; n = 12) or without (Macrosurgery; n = 16) a 3D exoscope between July 2018 and February 2021. The feasibility of performing all surgical steps with the 3D exoscope was evaluated. The Exoscope and Macrosurgery groups were compared for surgical time, vocal function outcomes, and complications. Questionnaires were completed by medical staff regarding the usefulness of medical education.ResultsNo intraoperative or postoperative complications occurred in either procedure. The operative time was similar in both groups. The vocal function outcomes were also comparable between the groups. Questionnaires revealed that the exoscope was useful in terms of sharing information on surgical procedures and anatomy, as well as functioning as an educational tool.ConclusionWhile this was a preliminary study, our results indicated that the exclusive use of the 3D exoscope was feasible for open approaches.Level of Evidence4 Laryngoscope, 134:1813–1819, 2024
Health Utility Score in Thai Patients with Chronic Rhinosinusitis: Pre‐ and Postoperative Analyses
Authors: Sangubol Melissa , Snidvongs Kornkiat , Lawpoolsri Saranath , Mongkolkul Kittichai , Kowatanamongkon Patlada , Chitsuthipakorn Wirach 
Published: October 2023

BackgroundEndoscopic sinus surgery (ESS) can increase the health utility score (HUS) of patients with chronic rhinosinusitis (CRS) who require the surgery. HUS varies depending on the geographical or living environment. HUS in CRS has never been evaluated in Thailand. The objective of this study was to evaluate the HUS of Thai patients with CRS before and after ESS through multiple approaches.MethodsThai patients (age ≥ 18) with CRS scheduled for ESS were enrolled in this prospective study. The preoperative (baseline) demographics, 22‐items sinonasal outcome test, endoscopic score, and CT score were recorded. The HUS was evaluated using four methods: the EuroQoL‐5 Dimension‐5 level (EQ‐5D‐5L), Visual Analog Scale (VAS), Standard gamble (SG), and Time trade‐off (TTO); at baseline, three months and six months post‐operation.ResultsData from 60 patients were analyzed. The mean baseline HUS scores by EQ‐5D‐5L, VAS, SG, and TTO were 0.75, 0.65, 0.79, and 0.85, respectively. The postoperative HUS significantly improved to 0.96, 0.91, 0.96, 0.97 at three months, and 0.97, 0.92, 0.97, and 0.98 at six months, respectively. ESS raised the HUS by 0.12–0.27 points. Among the four methods, VAS showed the lowest HUS at all time points.ConclusionThe preoperative HUS in Thai patients with CRS generally increased to near‐perfect values after the ESS. The increase in HUS reflecting the improved general quality of life, was demonstrated at three and up to six months after ESS.Level of Evidence4 Laryngoscope, 134:2070–2076, 2024