Swim-up of tammar wallaby (Macropus eugenii) spermatozoa in Biggers, Whitter and Whittingham (BWW) medium: maximisation of sperm motility, minimisation of impairment of sperm metabolism and induction of sperm hyperactivation.

A variety of media were compared for their ability to sustain the motility of tammar wallaby spermatozoa over an 8-h period following swim-up from coagulated semen. The study demonstrated that a modified Tyrode's solution, Biggers, Whitter and Whittingham medium (BWW) was significantly better than any of the other assessed media in supporting wallaby sperm motility. After 8h of incubation in BWW, motility was maintained at 79.3±9.3%, with 77.0±10.4% rapid and 65.7±8.7% progressively motile spermatozoa. By contrast, motility was <10% at the same 8-h time point in all of the other media assessed. After 2h of incubation in BWW, tammar spermatozoa consumed more oxygen than their counterparts in PBS (52.0±2.7 vs 75.0±6.6µL per 108 spermatozoa per 2h; P<0.001). Motility was not enhanced in any of these media by the addition of 5mM N-acetyl-D-glucosamine, the major energy substrate in wallaby semen. However, addition of dibutyryl cAMP and pentoxifylline in BWW resulted in the extremely rapid induction of hyperactivated motility in the entire sperm population. This burst of hyperactivated motility was entirely dependent on calcium in BWW and significantly inhibited by calmidazolium, a calmodulin inhibitor. A set of computer-assisted sperm analysis parameters were identified that permitted the accurate quantification of hyperactivation rates in this species. This is the first comparative analysis of media for harvesting and incubating marsupial spermatozoa and the first record of hyperactivated motility in any marsupial species.

Efficacy and safety of Huashi Baidu granule plus Nirmatrelvir-Ritonavir combination therapy in patients with high-risk factors infected with Omicron (B.1.1.529): A multi-arm single-center, open-label, randomized controlled trial.

BACKGROUND: Huashi Baidu granule (HSBD) and Paxlovid (Nirmatrelvir-Ritonavir) are antiviral Chinese patent medicine and western medicine specially developed for treating coronavirus disease 2019 (COVID-19). Their efficacy and safety in treating COVID-19 are still under investigated. PURPOSE: To assess and compare the efficacy and safety of HSBD, Paxlovid, and the combination in treating high-risk patients infected with SARS-CoV-2 Omicron. STUDY DESIGN: The study was a prospective single-center, open-label, randomized, controlled clinical trial conducted from April 18 to June 5, 2022. (ClinicalTrial.gov registration number: ChiCTR2200059390) METHODS: 312 severe patients aged 18 years and older infected with SARS-CoV-2 Omicron from Shuguang Hospital in Shanghai were randomly allocated to HSBD monotherapy (orally 137 g twice daily for 7 days, n = 105), Paxlovid monotherapy (orally 300 mg of Nirmatrelvir plus 100 mg of Ritonavir every 12 h for 5 days, n = 103), or combination therapy (n = 104). The primary outcome was SARS-CoV-2 nucleic acid negative conversion within 7-day treatment. The secondary outcome included hospital discharging conditions, severe conversion of symptom, and adverse events. RESULTS: Of 312 participants, 85 (82%) of 104 in combination therapy, 71 (68%) of 105 in HSBD monotherapy, and 73 (71%) of 103 in Paxlovid monotherapy had a primary outcome event. The hazard ratios of primary outcome were 1.37 (95% CI 1.03 - 1.84, p = 0.012) for combination versus HSBD, 1.28 (0.98-1.69, p = 0.043) for combination versus Paxlovid, and 0.88 (0.66-1.18, p = 0.33) for HSBD versus Paxlovid. There was no statistical difference of efficacy between HSBD and Paxlovid, while combination therapy exhibited more effective than either alone. For secondary outcomes, the hospital discharging rates within 7 days exhibited the significant increase in combination therapy than in HSBD or Paxlovid monotherapy (71% (74/104) vs 55% (58/105) vs 52% (54/103), p < 0.05). The risk of severe conversion of symptom showed no statistical significance among three interventions (1% (1/104) vs 3% (3/105) vs 3% (3/103), p > 0.05). No severe adverse events occurred among combination therapy and monotherapies in the trial. CONCLUSION: For patients with severe COVID-19, HSBD exhibits similar efficacy to Paxlovid, while combination therapy is more likely to increase the curative efficacy of Omicron variant than monotherapies, with few serious adverse events.

Differential effects of petroleum-based and bio-based microplastics on anaerobic digestion: A review.

The number of plastics is increasing owing to the rapid development of the plastics industry. Microplastics (MPs) are formed during the use of both petroleum-based plastics and newly developed bio-based plastics. These MPs are inevitably released into the environment and are enriched in wastewater treatment plant sludge. Anaerobic digestion is a popular sludge stabilization method for wastewater treatment plants. Understanding the potential impacts of different MPs on anaerobic digestion is critical. This paper provides a comprehensive review of the mechanisms of petroleum-based MPs and bio-based MPs in anaerobic digestion methane production and compares their potential effects on biochemical pathways, key enzyme activities, and microbial communities. Finally, it identifies problems that must be solved in the future, proposes the focus of future research, and predicts the future development direction of the plastics industry.

MicroRNA-145-5p regulates the proliferation of epithelial ovarian cancer cells via targeting SMAD4.

BACKGROUND: Epithelial ovarian cancer (EOC) is one of the most prevalent malignancies affecting females worldwide; however, its etiology mechanism remains unclear. In various malignancies, miR-145-5p is a widely accepted and versatile miRNA. Therefore, our research focused on exploring the activity and etiology of miR-145-5p in the modulation of metastasis, migration, and proliferation of EOC cells. The direct reactions between the 3'UTRs of SMAD4 mRNA and miR-145-5p were verified using dual luciferase reporter test. SKOV-3 cells were subsequently transfected using miR-145-5p mimics. Cell migration, death, and proliferation were evaluated using MTT, flow cytometry, and Transwell test. In addition, SMAD4 transcription and translation were evaluated using qRT-PCR and Western blot. RESULTS: We found that miR-145-5p expression was repressed prevalently in EOC tissues, apart from SMAD4 upregulation. Excessive miR-145-5p expression remarkably reinforced EOC cell death and repressed EOC cell proliferation. Furthermore, upregulated miR-145-5p expression noticeably repressed migration via MMP-2 and MMP-9 downregulation. Moreover, SMAD4 was downregulated via miR-145-5p transfection. The dual luciferase test revealed that miR-145-5p directly targeted SMAD4. CONCLUSIONS: Our research suggests that miR-145-5p serves as a malignancy repressor and exerts an essential impact on inhibiting malignancy generation and reinforcing EOC death via targeting SMAD4. MiR-145-5p application could serve as a promising strategy to treat EOC.

Tracing sperm acrosome differentiation in the testis and maturation in the epididymis of the tammar wallaby (Macropus eugenii) with a 45-kDa acrosome-membrane-associated protein.

A 45-kDa protein was originally extracted from a depression, where the acrosome is lodged, on the anterior end of the sperm nucleus of ejaculated wallaby spermatozoa. Using immunofluorescent and confocal microscopes, this study demonstrates that the 45-kDa protein is persistently localized to the sperm acrosome throughout the periods of spermiogenesis, spermiation, epididymal maturation and ejaculation in the tammar wallaby. The distribution of the 45-kDa protein is always on the perimeter of the acrosome and associated with the acrosomal membrane, so that changes in the shape of the 45-kDa immunofluorescent labelling mirror changes in the shape of the acrosome during its differentiation in the testis and epididymis. Thus, the 45-kDa protein may be used as a molecular marker to study the marsupial acrosome differentiation and to chart the events of testicular and epididymal maturation of the spermatozoa. Furthermore, the behaviour of the 45-kDa protein during the immunostaining process suggests that this protein is a largely insoluble and detergent-resistant protein and may play an important role in attachment of the acrosome to the nucleus during sperm formation, similar to those inner acrosomal-membrane-associated proteins that have been reported in eutherian spermatozoa.

An unusual abscisic acid and gibberellic acid synergism increases somatic embryogenesis, facilitates its genetic analysis and improves transformation in Medicago truncatula.

Somatic embryogenesis (SE) can be readily induced in leaf explants of the Jemalong 2HA genotype of the model legume Medicago truncatula by auxin and cytokinin, but rarely in wild-type Jemalong. Gibberellic acid (GA), a hormone not included in the medium, appears to act in Arabidopsis as a repressor of the embryonic state such that low ABA (abscisic acid): GA ratios will inhibit SE. It was important to evaluate the GA effect in M. truncatula in order to formulate generic SE mechanisms, given the Arabidopsis information. It was surprising to find that low ABA:GA ratios in M. truncatula acted synergistically to stimulate SE. The unusual synergism between GA and ABA in inducing SE has utility in improving SE for regeneration and transformation in M. truncatula. Expression of genes previously shown to be important in M. truncatula SE was not increased. In investigating genes previously studied in GA investigations of Arabidopsis SE, there was increased expression of GA2ox and decreased expression of PICKLE, a negative regulator of SE in Arabidopsis. We suggest that in M. truncatula there are different ABA:GA ratios required for down-regulating the PICKLE gene, a repressor of the embryonic state. In M. truncatula it is a low ABA:GA ratio while in Arabidopsis it is a high ABA:GA ratio. In different species the expression of key genes is probably related to differences in how the hormone networks optimise their expression.