Largazole Pharmacokinetics in Rats by LC-MS/MS

A highly sensitive and specific LC-MS/MS method for the quantitation of largazole thiol, the active species of the marine-derived preclinical histone deacetylase inhibitor, largazole (prodrug), was developed and validated. Largazole thiol was extracted with ethyl acetate from human or rat plasma along with the internal standard, harmine. Samples were separated on an Onyx Monolithic C18 column by a stepwise gradient elution with 0.1% formic acid in methanol and 0.1% aqueous formic acid employing multiple reaction monitoring (MRM) detection. Linear calibration curves were obtained in the range of 12.5–400 ng/mL with 200 µL of human plasma. The overall intra-day precision was from 3.87% to 12.6%, and the inter-day precision was from 7.12% to 9.8%. The accuracy at low, medium and high concentrations ranged from 101.55% to 105.84%. Plasma protein bindings of largazole thiol in human and rat plasma as determined by an ultrafiltration method were 90.13% and 77.14%, respectively. Plasma drug concentrations were measured by this LC-MS/MS method. The pharmacokinetics of largazole thiol in rats was studied following i.v. administration at 10 mg/kg and found to follow a two-compartment model. Largazole thiol was rapidly eliminated from systemic circulation within 2 h. The established LC-MS/MS method is suitable for the analysis of largazole thiol in human plasma, as well.     

Factors affecting porcine sperm mediated gene transfer

“Sperm mediated gene transfer” (SMGT) is based on the ability of sperm cells to bind exogenous DNA. The main objective of this study was to improve the production of transgenic pigs by SMGT. Taking into account that there is a lack of repeatability in studies of SMGT and that the mechanism of binding and internalization of exogenous DNA is a question that has not been solved, different factors involved in the production of transgenic animals by SMGT method were evaluated. Here we set out to: (1) evaluate the sperm capacity to bind exogenous DNA after DMSO treatment; (2) determine the location of the transgene–spermatozoa interaction; and (3) evaluate the efficiency of production of transgenic piglets by deep intrauterine artificial insemination (AI) with sperm incubated with DNA. The percentage of DNA binding was higher than 30% after 2 h of co-culture, but it was not affected by sperm treatment with DMSO (0.3% or 3%). The integrity of the sperm plasma membrane plays a critical role in DNA interaction, and altered plasma membranes facilitate interactions with exogenous DNA. DNA bound mainly to spermatozoa with reduced viability. DNA molecules were found to be mainly associated to the post-acrosomal region (61.9%). After deep intrauterine AI a total of 29 piglets were obtained, but none of them integrated the transgene. In conclusion, although it has been confirmed that DNA can associate with boar spermatozoa, the efficiency of producing transgenic pigs by AI was not confirmed by the present experiments, mainly due to a reduced DNA binding to functional spermatozoa.     

Avian regulation of crop and forest pests,a meta-analysis

BACKGROUND

Birds have been shown to reduce pest effects on various ecosystem types. This study aimed to synthesize the effect of birds on pest abundance, product damage and yield in agricultural and forest systems in different environments. Our hypothesis is that birds are effective pest regulators that contribute to a reduction in pest abundance, enhancement of yield quality and quantity and economic profit, and that pest regulation may depend on moderators such as the type of ecosystem, climate, pest, and indicator (ecological or economic).

RESULTS

We performed a systematic literature review of experimental and observational studies related to biological control in the presence and absence of regulatory birds. We retained 449 observations from 104 primary studies that were evaluated through qualitative and quantitative analyses. Of the 79 studies with known effects of birds on pest regulation, nearly half of the 334 observations showed positive effects (49%), 46% showed neutral effects, and very few (5%) showed negative effects. Overall effect sizes were positive (mean Hedges’ d = 0.38 ± 0.06). A multiple model selection retained only ecosystem and indicator types as significant moderators.

CONCLUSION

Our results support our hypothesis that there is a positive effect of avian control of pests for each analyzed moderator and this effect was significant for both ecological and economic indicators. Avian regulation of pests is a potential effective approach for environmentally friendly pest management that can reduce pesticide use regardless of the context of implementation. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Indirect fluorescent antibody techniques for demonstration of trout viruses and corresponding antibody

Two variations of the indirect fluorescent antibody technique (FAT) have been utilized in the work concerning two important virus diseases of trout, viral haemorrhagic septicaemia (VHS) and infectious pancreatic necrosis (IPN). A “two layer” indirect FAT allowed demonstration of the respective viruses in cell cultures and a “three layer” indirect FAT allowed demonstration of trout antibody to the viruses. Antibody, by means of the latter technique, could be demonstrated only in artificially immunized trout.     

MUSCULOSKELETAL LESIONS AND LAMENESS IN 121 HORSES WITH CARPAL SHEATH EFFUSION (1999–2010)

Equine carpal sheath effusion has multiple etiologies. The purpose of this retrospective study was to describe the prevalence of distinct musculoskeletal lesions lameness in a sample of horses with a clinical diagnosis of carpal sheath effusion. A total of 121 horses met inclusion criteria. Seventy‐four percent (89/121) of horses were lame at presentation; middle‐aged (9–18 years, 80%) and older (> 18 years, 85%) horses were lame more frequently than young horses (< 9 years, 44%). Ninety‐three percent (113/121) were diagnosed with osseous and/or soft tissue abnormalities. Of these 113 horses, 10 exhibited osseous abnormalities, whereas 111 were diagnosed with soft tissue lesions. Eighty‐four percent (93/111) of the soft tissue injuries extended from the caudodistal antebrachium to the palmar metacarpus. The superficial digital flexor tendon (98/111; 88%) and accessory ligament of the superficial digital flexor tendon (64/111; 58%) were the most commonly injured structures, with both structures affected in 41 (41/111; 37%) horses. Injuries within the caudodistal antebrachium included the superficial digital flexor musculotendinous junction (66), the accessory ligament of the superficial digital flexor tendon (64), and deep digital flexor muscle (21), in isolation or in combination with other structures. Increased echogenicity in the medial superficial digital flexor musculotendinous junction was detected in 40 horses and was significantly associated with increasing age (middle‐aged, 19/40; old, 18/40). Findings from this study indicated that age should be taken into consideration for horses presented with carpal sheath effusion and that adjacent structures within the caudodistal antebrachium should be included in evaluations.     

Genetics of tomato spotted wilt virus resistance coming from Lycopersicon peruvianum

New resistance sources coming from Lycopersicon peruvianum, especially those introgressed in UPV 32 line, are studied. UPV 32 resistance is controlled by a single gene. Resistance and dominance levels of this gene are conditioned by thrips transmission and isolate aggressiveness. A partial overcoming of resistance occurs due to the incomplete penetrance of the gene. Incomplete dominance of resistance also happens, which suggests gene dosage dependence. UPV 32 gene segregates independently of both Sw-5 gene and UPV 1 resistance gene, also coming from Lycopersicon peruvianum. It is proposed to name Sw-6 this new locus from UPV 32. Sw-5 gene and UPV 1 resistance gene show higher resistance than Sw-6. Heterozygotes for UPV 1 resistance gene were more resistant than heterozygotes for Sw-5. The lower dependence of UPV 1 resistance gene on the gene dosage effect makes it very useful for the development of commercial hybrids.