Effect of supplementation of cryoprotectant solution with hydroxypropyl cellulose for vitrification of bovine oocytes.

BACKGROUND: Successful cryopreservation of bovine oocytes is very important for research and commercial applications. However, the survival and development rate of vitrified-thawed (VT) oocytes are lower than those of non-vitrified-thawed (non-VT) oocytes. OBJECTIVE: To investigate the effect of adding hydroxypropyl cellulose (HPC) to the vitrification solution for bovine oocytes. MATERIALS AND METHODS: For vitrification, bovine metaphase II oocytes were pretreated with a solution containing 10% ethylene glycol supplemented with 0, 10, 50, or 100 ug/mL HPC for 5 min, exposed to a solution containing 30% ethylene glycol supplemented with 0, 10, 50, or 100 ug/mL HPC for 30 s, and then directly plunged into liquid nitrogen. RESULTS: The survival rate of oocytes was significantly higher in the 50 HPC group than in the 0, 10, and 100 HPC groups. The reactive oxygen species level was lower in the non-VT and 50 HPC groups than in the other groups. The mRNA levels of proapoptotic genes (Bax) were lower in the non-VT, 0, and 50 HPC groups than in the other groups. The mRNA levels of antiapoptotic genes (BCl2) were higher in the non-VT than in the other groups. The development rates of embryos (day 8) obtained via parthenogenetic activation (PA) were determined in the non-VT, 0 HPC, and 50 HPC groups. The cleavage rate was significantly higher in the non-VT group. CONCLUSION: Supplementation of vitrification solution with HPC improves the survival of VT bovine oocytes and the development capacity of embryos derived from these oocytes via PA. doi.org/10.54680/fr23110110212.

Validation of a scoring tool to predict drug-resistant pathogens in hospitalised pneumonia patients.

BACKGROUND: Health care-associated pneumonia (HCAP) affects a heterogeneous group of patients in frequent contact with health care systems. However, HCAP criteria poorly predict infection with drug-resistant (DR) pathogens. OBJECTIVE: To validate our previously reported risk-scoring model (predictive of DR pathogen infection) in patients admitted to hospital with pneumonia. DESIGN: We evaluated 580 patients admitted with culture-positive bacterial pneumonia. We identified risk factors, evaluated the risk-scoring model's capacity to predict infection by DR pathogens and compared the model's diagnostic accuracy with that of current HCAP criteria. RESULTS: DR pathogens were observed in 227/580 patients (39.1%). Of 269 HCAP patients, 153 (56.9%) were infected with DR pathogens. Overtreatment was more common in HCAP than in community-acquired pneumonia (58.7% vs. 41.2%, P < 0.001). Recent hospitalisation, admission from a long-term care facility, recent antibiotic treatment and tube feeding were independently associated with DR pathogens. For pathogen prediction, the risk-scoring model showed better diagnostic accuracy than HCAP criteria (area under receiver operating-characteristic curve = 0.723 vs. 0.673, P < 0.001). CONCLUSION: According to current HCAP criteria, half of the HCAP patients were treated unnecessarily with broad-spectrum antibiotics. Risk scoring by stratifying risk factors could improve the identification of patients likely to be infected with DR pathogens.

Aqueous extract of Schizandra chinensis fruit causes endothelium-dependent and -independent relaxation of isolated rat thoracic aorta.

An aqueous extract of Schizandra chinensis fruit (ScEx) has long been used to promote the vascular health of postmenopausal women in Korea. This study investigated the ability of ScEx to relax rat aorta constricted with norepinephrine (NE) and the mechanism(s) of such relaxation. ScEx induced partial, endothelium-dependent relaxation. In particular, the relaxation induced by lower concentrations of ScEx (0.1 and 0.3 mg/ml) was largely endothelium-dependent, and was essentially abolished by NG-nitro-L-arginine, methylene blue, 1H-[1,2,3] oxadiazole [4,4-a] quinoxalin-1-one, indomethacin, or ICI 182,780. The results indicate that the response to ScEx involves enhancement of the nitric oxide (NO)-cGMP system, and that it occurs via estrogen receptors. The magnitude of the inhibition with these treatments decreased with increasing ScEx concentration, however, indicating that other vasorelaxation mechanisms are involved, which depend on the ScEx concentration. Calcium concentration-dependent contraction curves in high potassium depolarization medium were shifted significantly to the right and downward after incubation with ScEx (0.3 and 1.0 mg/ml), implying that ScEx is also involved in inhibition of the extracellular calcium influx to vascular smooth muscle. These data demonstrate that ScEx caused both endothelium-dependent and -independent vasorelaxation, which may contribute to understanding the cardiovascular protective effect of ScEx.

Nitric oxide-mediated vasorelaxation by Rhizoma Ligustici wallichii in isolated rat thoracic aorta.

The vasorelaxant effect of Rhizoma Ligustici wallichii and its possible mechanism of action on the vasomotor tone of the rat thoracic aortic rings were examined in an organ bath. Chloroform extracts of Rhizoma Ligustici wallichii (Ch1LW) elicited a dose-dependent, transient, relaxing response in endothelium-intact rat aorta contracted with norepinephrine (NE). This relaxant effect was abolished by removal of the endothelium and also by pretreatment with nitric oxide synthase inhibitors. Neither a muscarinic receptor antagonist nor a cyclooxygenase inhibitor altered the Ch1LW-induced relaxation. Tetramethylpyrazine, derived from Rhizoma Ligustici wallichii as a potent vasodilating component, induced a complete relaxation in both endothelium-intact and denuded rat aortas contracted by NE, but nitric oxide synthase inhibitors did not affect the relaxation. Ch1LW-induced endothelium-dependent relaxation was mediated by nitric oxide released from the endothelium, and could be caused by component(s) other than tetramethylpyrazine.

Anti-HIV type 1 activity of 3'-fluoro-3'-deoxythymidine for several different multidrug-resistant mutants.

The objective of this work was to test the antiviral activity of a potent nucleoside reverse transcriptase inhibitor, 3'-fluoro-3'-deoxythymidine (FLT), on both a wild-type human immunodeficiency virus (HIV-1) isolate and multidrug-resistant HIV-1 patient isolates. Drug-resistant viral isolates were selected on the basis of four different categories of well-characterized and representative multidrug-resistant mutants. The isolates included three variants containing 151M alone or in combination; three variants containing 215Y and 41L, 67N, 184V, 210W, and 219N in combination; two insertion mutant viruses (69 + EA and 69 + SA); and two deletion mutant viruses (del67NG and del67GS), the latter two groups both also containing other significant mutations. The activity of FLT and AZT against these isolates was determined by drug susceptibility assays and by measuring viral antigen p24 by ELISA. The cytotoxicity of FLT and AZT was assessed in PHA-stimulated PBMCs. Development of resistant mutants under FLT pressure was attempted by passaging HIV-1 isolates in SupT1 cells and stepwise increasing the concentration of FLT. The multidrug-resistant mutant HIV-1 isolates exhibited 7-fold to >100-fold increased resistance to AZT, but showed IC(50) values for FLT of 0.0014-0.0168 microM, which were lower than or similar to that of wild type (0.0075 microM). The cellular cytotoxicities of FLT and AZT fell into a similar range in PBMCs. The development of HIV mutants resistant to FLT appeared to be slower than for other RT inhibitors. HIV isolates with mutations resulting in multidrug resistance had no evidence of resistance to FLT. FLT may be useful in salvage therapies for patients harboring resistant strains and a reassessment of its therapeutic potential seems required.

Effect of an oral dose of vitamin E on the vitamin E and cholesterol content of tissues of the vitamin E-deficient rat.

Weanling rats (female Sprague-Dawley) were fed until maturity a vitamin E-deficient diet or the deficient diet supplemented with 66 IU RRR-alpha-tocopheryl acetate/kg. Vitamin E, vitamin E quinone and total cholesterol levels in plasma, liver, paraovarian adipose tissue, lung, ovary and adrenal tissue were measured by high performance liquid chromatography. Vitamin E levels were greatly diminished, but cholesterol levels were unchanged in all tissues except adipose tissue of animals fed the deficient diet. Vitamin E-deficient animals received a single oral dose of 2 or 16.7 mg of RRR-alpha-tocopherol, and tissues were examined at 12 and 48 h. Plasma and liver formed a vitamin E pool that peaked at 12 h, had a high vitamin E/cholesterol ratio at 12 h and contained only trace amounts of vitamin E quinone. Adipose tissue, lung, ovary and adrenal concentrated vitamin E throughout the 48-h period, had low vitamin E/cholesterol ratios and contained small but significant amounts of vitamin E quinone. Vitamin E levels (micrograms/gram) at 48 h in lung, ovary and adrenal were higher than the vitamin E level in liver but the liver contained much more vitamin E (micrograms/organ) than the other tissues combined. Cholesterol levels (micrograms/gram) in plasma and liver decreased 45 to 55% in a dose- and time-dependent manner when a single oral dose of vitamin E was administered to deficient animals. Cholesterol levels in adipose tissue, lung and ovary were unchanged while the cholesterol level in adrenal increased 122% in a time-dependent manner with a single oral dose of vitamin E. These data show that a single oral dose of vitamin E has a profound effect on cholesterol levels in short-time experiments with the vitamin E-deficient rat. This rat model is appropriate for studies on the relationship between vitamin E and cholesterol metabolism in plasma, liver and the adrenal.