Efficacy of hysteroscopic levonorgestrel­releasing intrauterine device fixation in the treatment of adenomyosis: A cohort study.

The present study aimed to investigate the efficacy of hysteroscopic levonorgestrel-releasing intrauterine device (LNG-IUD) fixation surgery in the treatment of adenomyosis through a cohort study. The cohort study was performed at the Affiliated Jinhua Hospital of Wenzhou Medical University (Jinhua, China). A total of 31 women with adenomyosis were initially recruited from June 2020 to June 2022 and divided into an experimental group and a control group. The experimental group underwent hysteroscopic LNG-IUD fixation surgery and the control group underwent conventional implantation of the levonorgestrel-releasing intrauterine system. The assessed efficacy outcomes included the time of LNG-IUD expulsion, postoperative vaginal bleeding time, dysmenorrhea, and the menstrual blood loss score (MBLS). A total of 31 participants completed the research. The LNG-IUD expulsion rate was 6.25 and 60% (P<0.05) in the experimental and control group, respectively. The LNG-IUD in place time was 20.50 months (Q1, 15.75; Q3, 24.00) in the experimental group and 10.00 months (Q1, 6.50; Q3, 15.00) in the control group (P<0.05); the time of vaginal bleeding after surgery in the experimental and control groups were 12.50 days (9.25, 16.25) and 120.00 days (75.00, 120.00), respectively (P<0.05). Multiple-factor Cox regression analysis revealed that the LNG-IUD expulsion in patients with adenomyosis is associated with the hysteroscopic LNG-IUD fixation surgery [hazard ratio (HR), 1954.09], uterine cavity depth (HR, 16.63), MBLS (HR, 1.14), history of gonadotropin-releasing hormone agonist treatment in the previous 6 months (HR, 2.10), history of vaginal delivery (HR, 1.79) and history of cervical laceration (HR, 3.69). In conclusion, hysteroscopic LNG-IUD fixation reduces the rate of LNG-IUD expulsion, prolongs the time of LNG-IUD in the uterine cavity, reduces the time of postoperative vaginal bleeding, relieves the symptoms of dysmenorrhea and reduces the menstrual volume in the patients with adenomyosis. The present trial was retrospectively registered in the Chinese Clinical Trial Registry on 28th December 2023 (registration no. ChiCTR2300079233).

Pleomorphic rhabdomyosarcoma of the vagina: A case report.

BACKGROUND: Rhabdomyosarcoma (RMS) of the vagina in postmenopausal women is an extremely rare malignant tumor that was originally described as a unique group of soft tissue sarcomas originating from primitive mesenchymal cells. It was first reported in postmenopausal women in 1970, and fewer than 50 postmenopausal patients have been reported to date. CASE SUMMARY: A 68-year-old multiparous female was admitted to the hospital on October 11, 2023, with the chief complaint of a mass causing vaginal prolapse with incomplete urination that had persisted for 4 months. The vaginal mass was approximately the size of a pigeon egg; after lying down, the vaginal mass retracted. Complete resection was performed, and vaginal pleomorphic RMS was diagnosed based on pathology and immunohistochemical staining features. The patient is currently undergoing chemotherapy. The present study also reviewed the clinical, histological, and immunohistochemical features and latest treatment recommendations for vaginal RMS. Any abnormal vaginal mass should be promptly investigated through pelvic examination and appropriate imaging. The current initial treatment for vaginal RMS is biopsy and primary chemotherapy. CONCLUSION: When surgery is planned for vaginal RMS, an organ-preserving approach should be considered.

Teaching nursing students: As an umbrella review of the effectiveness of using high-fidelity simulation.

AIM: To conduct an umbrella review of the effectiveness of using high-fidelity simulation in nursing student teaching, thereby supporting continuous improvement in teaching practitioners' implementation of high-fidelity simulation intervention strategies. BACKGROUND: Several systematic reviews have investigated the effectiveness of high-fidelity simulation in nursing student teaching in recent years. However, conclusions vary and a systematic assessment is lacking. DESIGN: This review encompasses an umbrella review. METHODS: A search of PubMed, Embase, Cochrane Library, Web of Science and OVID databases was conducted to retrieve data on an umbrella review of high-fidelity simulation effectiveness in nursing student teaching from database inception to November 2023. The quality of the included systematic reviews was independently assessed by two reviewers using the AMSTAR 2 and PRISMA scales. Outcome indicators from the included systematic reviews were graded using the GRADE system. RESULTS: Twelve systematic reviews focusing on high-fidelity simulation effectiveness in nursing student teaching were included. Notably, all 12 systematic reviews exhibited very-low methodological quality, with 9 exhibiting some degree of reporting deficiencies, 2 exhibiting severe information deficiencies and 1 reporting relatively complete information. A total of 22 outcome indicators and 53 pieces of evidence were included. The results revealed 15 pieces of low-quality evidence and 38 pieces of very-low-quality evidence. Mounting evidence suggests that high-fidelity simulation teaching effectively enhances nursing students' theoretical performance, practical skills and various clinical comprehensive abilities, highlighting a positive teaching effect. However, further validation through high-quality, large-sample studies is warranted. CONCLUSION: The overall evidence quality of the current systematic reviews evaluating high-fidelity simulation effectiveness in nursing student teaching is low. Additionally, the methodological quality and the degree of reporting standardization require further improvement. Therefore, high-quality, large-sample randomized controlled trials are essential for further substantiating high-fidelity simulation effectiveness in nursing student teaching.

Reproductive outcomes following copper­containing intrauterine device after hysteroscopic lysis for intrauterine adhesions.

The present study aimed to investigate the reproductive outcomes of copper-containing intrauterine devices (IUDs) after hysteroscopic lysis in patients with mild to severe intrauterine adhesions (IUAs), according to the American Fertility Society (AFS) classification. Therefore, a prospective randomized controlled study was conducted at the Affiliated Jinhua Hospital of Wenzhou Medical University (Jinhua, China). A total of 173 women with IUAs were initially recruited between January 2020 and June 2021 and were then randomized to the copper-containing IUD group or the no barrier device group. Following hysteroscopic procedure, the fertility and obstetric outcomes were analyzed. Among the 173 patients enrolled, a total of 109 participants completed the study protocol. The results showed that AFS scores were not significantly different between the two groups prior to hysteroscopy. In addition, no statistically significant differences were recorded in pregnancy and live birth rates between the copper-containing IUD and no barrier device groups. Overall, the results of the current study indicated that the copper-containing IUDs had no positive effect on pregnancy and live birth rates in patients with mild to severe IUAs after hysteroscopic adhesiolysis. The present trial was retrospectively registered in the Chinese Clinical Trial Registry on 28th December 2023 (registration no. ChiCTR2300079233).

[A case of high-pressure injection injury of hand complicated with deep chemical burn caused by industrial cement].

This article reports a patient with extensive high-pressure injection injury of the hand combined with deep chemical burn caused by high-pressure injection of industrial cement materials was diagnosed and treated in the Department of Hand Surgery, Xiaolan Hospital Affiliated to Southern Medical University in 2022. The nerves, tendons and blood vessels of the left hand were involved, and the ulnar skin of the left thumb was extensively necrosis, and a large number of extensive cement foreign bodies remained under the skin. Part of the cement was inserted into the joint capsule of the interphalangeal joint. After emergency surgical treatment, the patient was saved successfully, and the wound healed well without chemical poisoning and other related complications, which created conditions for the second stage of flap repair.

Areas with high HIV prevalence: A spatial analysis of nationwide claims data in Germany.

OBJECTIVE: We aimed to identify spatial clusters of high HIV prevalence in Germany. METHODS: Using nationwide outpatient claims data comprising information of about 88% of the total German population (N = 72 041 683), we examined spatial variations and spatial clusters of high HIV prevalence at the district level (N = 401). People with HIV were identified using the International Statistical Classification of Diseases and Related Health Problems, Tenth Revision (ICD-10 codes) B20, B22, and B24 (HIV disease) documented as 'confirmed'. RESULTS: Among 72 041 683 people with statutory health insurance in Germany in 2021, 72 636 had diagnosed HIV, which corresponds to a prevalence of 101 per 100 000 individuals (0.10%). Of these, 56 895 were males (78%). At a district level, the HIV prevalence varied by a factor of 32 between 13 in a rural district in Bavaria and 417 per 100 000 individuals in the German capital, Berlin. The spatial autocorrelation coefficient was 0.24 (p < 0.0001, Global Moran's I). Several high-prevalence spatial clusters of different sizes were identified, mostly located in western Germany. The largest cluster comprised eight districts in the southern part of Hesse, including the city of Frankfurt and the city of Mainz in Rhineland-Palatinate. The second cluster consisted of four districts in North Rhine-Westphalia, including the cities of Cologne and Düsseldorf. Two districts in southern Germany (Mannheim and Ludwigshafen) formed the third cluster. Only urban districts were observed in spatial clusters of high HIV prevalence. CONCLUSIONS: The current study identified for the first time spatial clusters with high HIV prevalence in Germany. This understanding is of particular importance when planning the general and specialized medical care of patients with HIV and to support preventive measures.

Evaluation of high dose N- Acetylcysteine on airway inflammation and quality of life outcomes in adults with bronchiectasis: A randomised placebo-controlled pilot study.

BACKGROUND: High dose N acetylcysteine (NAC), a mucolytic, anti-inflammatory and antioxidant agent has been shown to significantly reduce exacerbations, and improve quality of life in placebo controlled, double blind randomised (RCT) studies in patients with COPD, and in an open, randomised study in bronchiectasis. In this pilot, randomised, double-blind, placebo-controlled study, we wished to investigate the feasibility of a larger clinical trial, and the anti-inflammatory and clinical benefits of high dose NAC in bronchiectasis. AIMS: Primary outcome: to assess the efficacy of NAC 2400 mg/day at 6 weeks on sputum neutrophil elastase (NE), a surrogate marker for exacerbations. Secondary aims included assessing the efficacy of NAC on sputum MUC5B, IL-8, lung function, quality of life, and adverse effects. METHODS: Participants were randomised to receive 2400 mg or placebo for 6 weeks. They underwent 3 visits: at baseline, week 3 and week 6 where clinical and sputum measurements were assessed. RESULTS: The study was stopped early due to the COVID pandemic. In total 24/30 patients were recruited, of which 17 completed all aspects of the study. Given this, a per protocol analysis was undertaken: NAC (n = 9) vs placebo (n = 8): mean age 72 vs 62 years; male gender: 44% vs 50%; baseline median FEV11.56 L (mean 71.5 % predicted) vs 2.29L (mean 82.2% predicted). At 6 weeks, sputum NE fell by 47% in the NAC group relative to placebo (mean fold difference (95%CI: 0.53 (0.12,2.42); MUC5B increased by 48% with NAC compared with placebo. Lung function, FVC improved significantly with NAC compared with placebo at 6 weeks (mean fold difference (95%CI): 1.10 (1.00, 1.20), p = 0.045. Bronchiectasis Quality of life measures within the respiratory and social functioning domains demonstrated clinically meaningful improvements, with social functioning reaching statistical significance. Adverse effects were similar in both groups. CONCLUSION: High dose NAC exhibits anti-inflammatory benefits, and improvements in aspects of quality of life and lung function measures. It is safe and well tolerated. Further larger placebo controlled RCT's are now warranted examining its role in reducing exacerbations.

Reconstructing a long-term water storage-based drought index in the Yangtze River Basin.

Drought is a prolonged dry period in the natural climate cycle, and is one of the most costly weather events. The Gravity Recovery and Climate Experiment (GRACE) derived terrestrial water storage anomalies (TWSA) have been widely used to assess drought severity. However, the relatively short cover period of GRACE and GRACE Follow-On limit our knowledge about the characterization and evolution of drought over decades time scale. This study proposes a standardized GRACE reconstructed TWSA index (SGRTI) to assess the drought severity based on a statistical reconstruction method calibrated by GRACE observations. Results show that the SGRTI correlates well with 6-month scale SPI and SPEI, with correlation coefficients reaching 0.79 and 0.81 in the YRB from 1981 to 2019. Soil moisture can capture drought condition like the SGRTI, while cannot further reflect deeper water storage depletion. The SGRTI is also comparable to the SRI and in-situ water level. As a case study for the Yangtze River Basin, its three sub-basins experience more frequent droughts, shorter drought duration, and lower severity drought, as identified by SGRTI during 1992-2019 relative to 1963-1991. The presented SGRTI in this study can provide a valuable supplement to the drought index before the GRACE era.

TRANSCRIPTOME ANALYSIS REVEALED THE MOLECULAR SIGNATURES OF CISPLATIN-FLUOROURACIL COMBINED CHEMOTHERAPY RESISTANCE IN GASTRIC CANCER.

Gastric cancer (GC) is among the top five malignant tumors worldwide in terms of morbidity and death. Chemotherapy is the primary treatment for unresectable or advanced postoperative GC. Chemotherapy resistance developed against cisplatin-fluorouracil (CF) combined chemotherapy is one of the most common clinical issues in patients with GC, leading to poor prognosis. Two different methods were used to analyze GSE14210, and two gene sets were obtained. The first method involved performing the traditional difference analysis (adjusted p<0.05, |log2FC|>=1) by Network Analyst to obtain gene set 1, followed by conducting gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis on the obtained gene set. The second method involved using iDEP to make the weighted gene co-expression network analysis (WGCNA) and performing GO and KEGG enrichment analysis, to obtain gene set 2. Thereafter, the STRING database and Cytoscape were used to construct Protein-Protein Interaction (PPI) networks, screen core clusters, and hub genes of the two gene sets. Furthermore, the hub genes were verified in GSE14210 by the survival analysis method of the Kaplan-Meier plotter database. Finally, we analyzed the mRNA expression of the hub genes by UALCAN and the protein expression of the same by Human Protein Atlas (HPA). Three real hub genes with the same mRNA expression as that of protein were identified, including CENPB, MTA1, and GCNT3. Finally, we performed single gene GO and KEGG enrichment analyses to explore the possible mechanisms of action of these three genes. The mRNA and protein expressions of CENPB, MTA1, and GCNT3 were upregulated in CF-resistant GC patients, and they were significantly associated with bad overall survival (OS). CENPB, MTA1 and GCNT3 are expected to be biomarkers with promising clinical applications as potential therapeutic targets for patients with refractory GC treated with CF combined chemotherapy.

Clinical Diagnosis of Cervical Lesions Combined with Programmed Death Ligand and miR-124 Detection in Peripheral Blood.

Objective: This study aims to analyze the expression of colposcopy combined with PD-L1 (programmed death ligand-1) and miR-124 (microRNA-124) in CC (cervical cancer) and CIN (cervical precancerous lesions), providing insights for clinical screening and diagnosis of these conditions. Method: A total of 60 patients with suspicious cervical lesions were selected from the gynecological clinic at Jinhua People's Hospital between June 2021 and December 2021. The patients were divided into three groups: LSIL (low-grade squamous intraepithelial lesions), HSIL (high-grade squamous intraepithelial lesions), and no SIL group, with 20 cases per group. This sample distribution ensures a comprehensive representation of different lesion severities. Pathological tissues were collected from each group for immunohistochemistry analysis to assess PD-L1 expression. Peripheral blood samples were also obtained from the patients for PCR analysis to evaluate miR-124 expression. These techniques allowed us to examine the expression levels of PD-L1 and miR-124 in the samples accurately. Result: The HSIL group exhibited a higher rate of positive PD-L1 expression compared to the LSIL and no lesion groups. Additionally, the expression level of miR-124 was lower in the HSIL group compared to the LSIL and no lesion groups (P < .05). Statistical measures such as means, standard deviations, and P values were used to quantify these differences, providing a more comprehensive understanding of the results. Conclusions: Combining colposcopy results with the expression of PD-L1 and miR-124 can effectively evaluate precancerous lesions of cervical cancer. This combined approach holds significant clinical implications by potentially enhancing early detection, diagnosis, and treatment strategies for CC and CIN. Further research in this area may lead to improved patient outcomes and contribute to the development of targeted therapies.

Is catalase an effective additive to alleviate oxidative stress during cryopreservation of zebrafish sperm at the repository level?

The goal of this study was to investigate whether supplementation of cryoprotective medium with catalase (CAT), an antioxidation enzyme, is efficient for zebrafish sperm cryopreservation from the viewpoint of high-throughput genetic repository operations. Three cryoprotectants (10%, v/v), dimethylacetamide (DMA), dimethylformamide (DMF), and methanol were used. The objectives were to evaluate the effects of CAT on sperm motility, plasma membrane integrity, and concentration for: 1) fresh sperm at equilibration up to 60 min; 2) post-thaw sperm after cooling at 10, 20, and 40 °C/min), and 3) post-thaw fertilization and embryo survival rates. Catalase addition did not improve sperm motility, regardless of the cryoprotectants added. After 10-min exposure to DMA or methanol, membrane integrity was significantly decreased (70-75%) compared to controls. With catalase, sperm cells maintained membrane integrity and after 50 min equilibration, cell concentrations were maintained with CAT compared to cryoprotectant-only test groups. However, after cryopreservation and thawing, CAT did not affect the outcome of motility, membrane integrity, cell concentration, fertilization, or embryo survival assays. Analysis of cooling rates also indicated that CAT did not affect 3-hpf fertilization or 24-hpf survival rates. Overall, addition of CAT could provide some protection of sperm from oxidative stress before freezing, but not after thawing. We propose that decisions concerning routine use of CAT for repositories, especially those handling tens of thousands of frozen samples per year, would depend on whether efficient high-throughput operation, or specific research questions are programmatic goals.

Temporal and Concentration Effects of Methanol on Cryopreservation of Zebrafish (Danio rerio) Sperm.

We evaluated the cryoprotective effects of methanol on zebrafish sperm at different concentrations, exposure times, and stages during cryopreservation. Samples were collected by crushing of dissected testes or abdominal stripping. After exposure to 0%, 2%, 5%, 8%, and 10% methanol for 0-11 min, fresh sperm (1 × 106 cells/mL) did not show changes in plasma membrane integrity (measured by flow cytometer), but cell size changes (light scatter) were observed after exposure to 8% or 10%. After exposure for 0-60 min, fresh sperm (1 × 108 cells/mL) did not show significant changes in survival or membrane integrity. Sperm cryopreserved in 5%, 8%, and 10% methanol showed high post-thaw survival, in 5% and 8% showed high post-thaw motility, and in 5% showed highest post-thaw membrane integrity compared to other concentrations between 0% and 10%. Within 0-60 min after thawing, no significant differences in cell survival and membrane integrity were found for any concentration (p ≥ 0.269). Comparison of 5% and 8% methanol for dissected testes (n = 20) revealed no difference in post-thaw motility, membrane integrity, cell survival, fertilization, or hatching, embryo viability; for stripped sperm (n = 10), no differences were observed in post-thaw membrane integrity, fertilization, and embryo viability, however, higher motility and survival were detected in 5% than in 8% methanol. Thus, a concentration of 5% methanol seems most suitable for cryopreserving zebrafish sperm based on post-thaw survival and motility.

Effect of miR-10a on sepsis-induced liver injury in rats through TGF-ß1/Smad signaling pathway.

OBJECTIVE: To observe the effect of the micro ribonucleic acid (miR)-10a on sepsis-induced liver injury in rats through the transforming growth factor-b1 (TGF-b1)/Smad signaling pathway. MATERIALS AND METHODS: The rat model of sepsis was established via cecal ligation and puncture, in which miR-10a was overexpressed and silenced using liposome transfection. The rats were randomly divided into miR-10a mimics group (Mimics group, n=10) and miR-10a inhibitors group (Inhibitors group, n=10), and the sham operation group (Sham group, n=10) was also set up. The transfection efficiency of miR-10a in liver tissues in each group was detected via quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), the serum liver function indexes aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were determined. Moreover, the content of the serum reactive oxygen species (ROS), glutathione (GSH), and GSH peroxidase (GSH-Px) was determined using enzyme-linked immunosorbent assay (ELISA). The content of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and myeloperoxidase (MPO) in liver tissues was detected, and the pathological changes in liver tissues were observed through hematoxylin-eosin (HE) staining. Finally, the expression levels of cytochrome P4502E1 (CYP2E1) and TGF-b1/Smad signaling pathway genes and proteins in liver tissues were detected via qRT-PCR and Western blotting. RESULTS: The expression of miR-10a was significantly increased in Mimics group (p<0.05) and extremely low in the Inhibitors group (p<0.05). In Mimics group, the levels of serum AST, ALT, and LDH were significantly increased (p<0.05), the content of ROS, TNF-α, IL-6, and MPO was substantially increased (p<0.05), while that of GSH and GSH-Px notably declined (p<0.05). According to the HE staining results, the liver cells were orderly arranged in the Inhibitors group, and they were disorderly arranged with more inflammatory cells in the Mimics group. The results of the gene and protein assays showed that the expression levels of CYP2E1, TGF-b1, and Smad2 in Mimics group were markedly higher than those in the Sham group (p<0.05), while they displayed the opposite trends in the Inhibitors group (p<0.05). CONCLUSIONS: Silencing miR-10a can inhibit the occurrence of sepsis-induced liver injury in rats by downregulating the TGF-ß1/Smad pathway.

Learning Desire Is Predicted by Similar Neural Processing of Naturalistic Educational Materials.

Naturalistic stimuli can elicit highly similar brain activity across viewers. How do naturalistic educational materials engage human brains and evoke learning desire? Here, we presented 15 audiovisual course clips (each lasting ∼120 s) to university students and recorded their neural activity through electroencephalography. Upon finishing all the video viewings, subjects ranked 15 courses in order of learning desire and reported the reasons for high learning desire (i.e., "value" and "interest"). The brain activity during the video viewing was measured as the neural similarity via intersubject correlation (ISC), that is, correlation between each subject's neural responses and those of others. Based on averaged learning desire rankings across subjects, course clips were classified with high versus medium versus low motivational effectiveness. We found that the ISC of high effective course clips was larger than that of low effective ones. The ISC difference (high vs low) was positively associated with subjects' learning desire difference (high vs low). Such an association occurred when viewing time accumulated to ∼80 s. Moreover, ISC was correlated with "interest-based" rather than "value-based" learning desire. These findings advance our understanding of learning motivation via the neural similarity in the context of on-line education and provide potential neurophysiological suggestions for pedagogical practices.

A Strategy for Sperm Cryopreservation of Atlantic Salmon, Salmo salar, for Remote Commercial-scale High-throughput Processing.

Sperm cryopreservation is an essential tool for long-term storage of genetic resources for aquaculture fishes. The goal of this study was to develop an efficient and streamlined protocol for high-throughput processing for sperm cryopreservation in Atlantic salmon, Salmo salar. The objectives were to evaluate: 1) osmolality of blood serum for determining extender osmolality; 2) effects of extenders for fresh sperm dilution and refrigerated storage; 3) effects of methanol and dimethyl sulfoxide (DMSO) on fresh sperm motility, and 4) motility and fertility after thawing. In this study, sperm samples were collected at a hatchery site in Canada, and shipped to a freezing site located 2200 miles (3550 km) away in the United States. Evaluation of three extenders indicated that Mounib solution was suitable for diluting dry sperm for sample processing. Ten percent of methanol or DMSO was less toxic to sperm cells than was 15% within 30 min. Further testing with methanol at 5, 10, and 15%, and sperm solution:extender dilutions (v:v) of 1:1, 1:3, 1:19 (at concentrations of ~5×107; 3×108, and 1×109 cells/mL) indicated that methanol at 5% and 10% showed less toxicity to fresh sperm within 1 hr at sperm: extender dilutions of 1:1 and 1:3. Post-thaw motility of sperm cryopreserved with 10% methanol was significantly higher than that with 10% DMSO, and fertility reflected those results (0-1% in DMSO vs. 38-55% in methanol). Further evaluation of sperm cryopreservation with 10 and 15% methanol at sperm dilution ratios of 1:1, 1:3, 1:19 indicated post-thaw motility in 10% methanol was significantly higher than that in 15% methanol, and post-thaw fertility in 10% methanol at 1:1 and 1:3 dilution ratios had fertilization rates similar to that of fresh sperm controls. Sperm samples from 12 males cryopreserved with 10% methanol showed male-to-male variation in post-thaw motility (0-36%). Overall, a simplified standard protocol was established for cryopreservation of shipped sperm of Atlantic salmon using extender without egg yolk and yielded satisfactory post-thaw motility and fertilization rates. This procedure can be readily adopted by aquaculture facilities to take advantage of high-throughput cryopreservation capabilities at remote service centers. Most importantly, this approach lays the groundwork for an alternative commercial model for commercial-scale production, quality control and development of industrial standards. Control of male variability and sperm quality remain important considerations for future work.

Vitrification as an Alternative Approach for Sperm Cryopreservation in Marine Fishes.

The Southern Flounder Paralichthys lethostigma is a high-value species and a promising aquaculture candidate. Because sperm volume can be limited in this species (<500 µL), new sperm cryopreservation methods need to be evaluated. Vitrification is an alternative to conventional slow-rate freezing, whereby small volumes are cryopreserved at high cooling rates (>1,000°C/min). The goal of this work was to develop a standardized approach for vitrification of Southern Flounder sperm. The specific objectives were to (1) evaluate thawing methods and vitrification solutions, (2) evaluate the postthaw membrane integrity of sperm vitrified in different cryoprotectant solutions, (3) examine the relationship between membrane integrity and motility, and (4) evaluate the ability of vitrified sperm to fertilize eggs. From the vitrification solutions tested, the highest postthaw motility (28 ± 9% [mean ± SD]) and membrane integrity (11 ± 4%) was observed for 20% ethylene glycol plus 20% glycerol. There was no significant difference in postthaw motility of sperm thawed at 21°C or at 37°C. Fertilization from vitrified sperm in one trial yielded the same fertilization rate (50 ± 20%) as the fresh sperm control, while the sperm from the other two males yielded 3%. This is the first report of fertilization by vitrified sperm in a marine fish. Vitrification can be simple, fast, inexpensive, performed in the field, and, at least for small fishes, offers an alternative to conventional cryopreservation. Because of the minute volumes needed for ultrarapid cooling, vitrification is not presently suited as a production method for large fishes. Vitrification can be used to reconstitute lines from valuable culture species and biomedical models, conserve mutants for development of novel lines for ornamental aquaculture, and transport frozen sperm from the field to the repository to expand genetic resources.

3-D printing provides a novel approach for standardization and reproducibility of freezing devices.

Cryopreservation has become an important and accepted tool for long-term germplasm conservation of animals and plants. To protect genetic resources, repositories have been developed with national and international cooperation. For a repository to be effective, the genetic material submitted must be of good quality and comparable to other submissions. However, due to a variety of reasons, including constraints in knowledge and available resources, cryopreservation methods for aquatic species vary widely across user groups which reduces reproducibility and weakens quality control. Herein we describe a standardizable freezing device produced using 3-dimensional (3-D) printing and introduce the concept of network sharing to achieve aggregate high-throughput cryopreservation for aquatic species. The objectives were to: 1) adapt widely available polystyrene foam products that would be inexpensive, portable, and provide adequate work space; 2) develop a design suitable for 3-D printing that could provide multiple configurations, be inexpensive, and easy to use, and 3) evaluate various configurations to attain freezing rates suitable for various common cryopreservation containers. Through this approach, identical components can be accessed globally, and we demonstrated that 3-D printers can be used to fabricate parts for standardizable freezing devices yielding relevant and reproducible cooling rates across users. With standardized devices for freezing, methods and samples can harmonize into an aggregated high-throughput pathway not currently available for aquatic species repository development.

Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development.

Cryopreservation in aquatic species in general has been constrained to research activities for more than 60 years. Although the need for application and commercialisation pathways has become clear, the lack of comprehensive quality assurance and quality control programs has impeded the progress of the field, delaying the establishment of germplasm repositories and commercial-scale applications. In this review we focus on the opportunities for standardisation in the practices involved in the four main stages of the cryopreservation process: (1) source, housing and conditioning of fish; (2) sample collection and preparation; (3) freezing and cryogenic storage of samples; and (4) egg collection and use of thawed sperm samples. In addition, we introduce some key factors that would assist the transition to commercial-scale, high-throughput application.

Metabolic syndrome, dyslipidemia and prostate cancer recurrence after primary surgery or radiation in a veterans cohort.

BACKGROUND: Metabolic syndrome (MetS) has been hypothesized to be associated with cancer, including prostate cancer (PCa), but the relationship is not well characterized. We analyze the relationship between MetS features and localized PCa recurrence after treatment. METHODS: Men having primary treatment for localized PCa were included from a multi-site regional veteran network. Recurrence was defined as nadir PSA +2 ng ml(-1) (radiation) or PSA⩾0.2 ng ml(-1) (prostatectomy). MetS was based on consensus professional society guidelines from the American Heart Association and International Diabetes Federation (three of: hypertension >130/85 mm Hg, fasting blood glucose ⩾100 mg dl(-1), waist circumference >102 cm, high-density lipoprotein <40 mg dl(-1), triglycerides ⩾150 mg dl(-1)). Closely related abnormality in low-density lipoprotein (LDL; >130 mg dl(-1)) was also examined. Analysis of PCa recurrence risk included multivariable Cox proportional hazards regression with propensity adjustment. RESULTS: Of the 1706 eligible men, 279 experienced recurrence over a median follow-up period of 41 months (range 1-120 months). Adjustment variables associated with PCa recurrence included: index PSA, Gleason, and tumor stage. Independent variables of interest associated with PCa recurrence were hyperglycemia and elevated LDL. Elevated LDL was associated with PCa recurrence (multivariable hazard ratio (HR) 1.34, 95% confidence interval (CI) 1.03, 1.74; propensity adjusted HR 1.33, 95% CI 1.03, 1.72). There was also an association between impaired fasting glucose and PCa recurrence in (multivariable HR 1.54, 95% CI 1.10, 2.15; propensity adjusted HR 1.41, 95% CI 1.01, 1.95). MetS was not associated with PCa recurrence (multivariable: HR 0.96, 95% CI 0.61, 1.50; propensity adjusted HR 1.04, 95% CI 0.67, 1.62). CONCLUSIONS: PCa recurrence is not associated with MetS but is associated with elevated LDL and impaired fasting glucose. If confirmed, these data may help provide modifiable targets in preventing recurrence of PCa.

Simulation modeling of high-throughput cryopreservation of aquatic germplasm: a case study of blue catfish sperm processing.

Emerging commercial-level technology for aquatic sperm cryopreservation has not been modeled by computer simulation. Commercially available software (ARENA, Rockwell Automation, Inc. Milwaukee, WI) was applied to simulate high-throughput sperm cryopreservation of blue catfish (Ictalurus furcatus) based on existing processing capabilities. The goal was to develop a simulation model suitable for production planning and decision making. The objectives were to: 1) predict the maximum output for 8-hr workday; 2) analyze the bottlenecks within the process, and 3) estimate operational costs when run for daily maximum output. High-throughput cryopreservation was divided into six major steps modeled with time, resources and logic structures. The modeled production processed 18 fish and produced 1164 ± 33 (mean ± SD) 0.5-ml straws containing one billion cryopreserved sperm. Two such production lines could support all hybrid catfish production in the US and 15 such lines could support the entire channel catfish industry if it were to adopt artificial spawning techniques. Evaluations were made to improve efficiency, such as increasing scale, optimizing resources, and eliminating underutilized equipment. This model can serve as a template for other aquatic species and assist decision making in industrial application of aquatic germplasm in aquaculture, stock enhancement, conservation, and biomedical model fishes.