Differential immunolocalization between lingual antimicrobial peptide and lactoferrin in mammary gland of dairy cows

Lingual antimicrobial peptide (LAP), one of the β-defensins in bovines, and lactoferrin (LF) are synthesized in mammary epithelium and have bactericidal and bacteriostatic functions. However, it is not known whether they have similar expression patterns. Therefore, the present study was undertaken to compare (1) immunolocalization of LAP and LF in the mammary gland and (2) changes in concentration of these two components in milk after lipopolysaccharide (LPS) challenge. Bovine mammary tissues without LPS challenge were collected and their sections were immunostained with antibodies to LAP or LF. Milk from our previous study was collected every hour up to 12h and twice daily from d 1 to 7 after LPS challenge (the day of infusion was considered as d 0). These milk samples were measured for LAP but not LF in our previous report. Therefore, concentration of LF was measured by enzyme immunoassay in the present study. Epithelial cells of some alveoli showed immunopositive reaction for LF, but negative for LAP. Conversely, some alveoli were LAP positive in their epithelial cells but LF negative. Many alveoli had immunoreactions for neither LAP nor LF. The concentration of LAP in milk was elevated significantly at 3h after LPS infusion compared with pre-infusion values and remained at a high level until 12h. However, LF concentration in milk remained low at d 0 and increased at d 2. These results suggest that LAP and LF were mostly differentially localized in the alveolar epithelium in mammary glands. The different spatial expressions between them may be associated with their different temporal expression mechanisms.     

Immunolocalization of lingual antimicrobial peptide (LAP) in the bovine mammary gland

Lingual antimicrobial peptide (LAP), a member of the β-defensin family in cows, is involved in the innate immune system and plays a crucial role in killing a large variety of microorganisms. The aim of the present study was to demonstrate the immunolocalization of LAP in the mammary glands of cows. A LAP antibody was raised in a rabbit by immunity with a synthetic 11 amino acid sequence out of a 42-amino acid sequence of the mature form of LAP. The specificity of the LAP antibody was checked using a competitive immunoassay and Western blotting. Paraffin sections of the mammary gland were immunostained with LAP antibody. In the competitive immunoassay, an increase of synthetic LAP concentration suppressed the optical density. Western blotting analysis for LAP revealed the presence of the LAP peptide in mammary alveolar tissue. When the mammary gland was immunostained with LAP antibody, epithelial cells of both infected and non-infected alveoli were immunopositive. These results indicate that LAP is localized in the epithelium of non-infected as well as infected alveolus in the mammary gland in cows.     

Effects of fallen snow on the voluntary intake and grazing behavior of Hokkaido native horses in winter woodland with underlying Sasa senanensis

The intake, digestibility, and grazing behavior of Hokkaido native horses were investigated in winter woodland with underlying Sasa senanensis. Twenty Hokkaido native horses were grazed in a woodland (8.52 ha) for 14 days in December (snowy; 20 cm of snow fall) and in November (non‐snowy) of the next year. Three mares (360 kg of average bodyweight) were used in each experiment. The fecal collection, used to investigate the intake and digestibility by the double‐indicator method, and behavioral observations were carried out for 24 h on the 6th day in each experiment. During the snowy and non‐snowy periods, the dry matter intakes of sasa foliage were 7.3 and 9.6 kg/d, respectively, which is 2.1 and 2.7% when expressed as a percentage of bodyweight. The apparent digestibility of nutrients and energy released during the snowy and non‐snowy periods were similar. The proportions of digestible energy intakes to maintenance requirement in the mares were 119 and 164% during the snowy and non‐snowy periods, respectively. Their bodyweight remained constant during the non‐snowy period; however, a slight loss in their bodyweight was observed during the snowy period. The duration of grazing on sasa foliage was 416 and 544 min/d during the snowy and non‐snowy periods, respectively, the daily grazing area of the mares during these periods was 2.0 and 5.1 ha, respectively. In other words, the area during the snowy period was smaller than that during the non‐snowy period (P < 0.05). Fallen snow, even with a depth of approximately 20 cm, inhibited the grazing behavior and intake of sasa foliage by Hokkaido native horses grazing in winter woodland pasture.     

Life in the fast lane: Revisiting the fast growth—High survival paradigm during the early life stages of fishes

Early life survival is critical to successful replenishment of fish populations, and hypotheses developed under the Growth-Survival Paradigm (GSP) have guided investigations of controlling processes. The GSP postulates that recruitment depends on growth and mortality rates during early life stages, as well as their duration, after which the mortality declines substantially. The GSP predicts a shift in the frequency distribution of growth histories with age towards faster growth rates relative to the initial population because slow-growing individuals are subject to high mortality (via starvation and predation). However, mortality data compiled from 387 cases published in 153 studies (1971–2022) showed that the GSP was only supported in 56% of cases. Selection against slow growth occurred in two-thirds of field studies, leaving a non-negligible fraction of cases showing either an absence of or inverse growth-selective survival, suggesting the growth-survival relationship is more complex than currently considered within the GSP framework. Stochastic simulations allowed us to assess the influence of key intrinsic and extrinsic factors on the characteristics of surviving larvae and identify knowledge gaps on the drivers of variability in growth-selective survival. We suggest caution when interpreting patterns of growth selection because changes in variance and autocorrelation of individual growth rates among cohorts can invalidate fundamental GSP assumptions. We argue that breakthroughs in recruitment research require a comprehensive, population-specific characterization of the role of predation and intrinsic factors in driving variability in the distribution and autocorrelation of larval growth rates, and of the life stage corresponding to the endpoint of pre-recruited life.     

Minimally invasive proximal interphalangeal joint arthrodesis using a locking compression plate and tissue engineering in horses: A pilot study

This pilot study assessed the efficacy of 2 minimally invasive techniques for proximal interphalangeal (PIP) joint arthrodesis in horses. The PIP joints of both forelimbs (n = 6) were stabilized with locking compression plates (LCP) using a minimally invasive technique (LCP technique). Subsequently, for 1 randomly selected PIP joint of each horse, surgical drilling (SurD) was performed and tissue engineering (TE) was applied (LCP/SurD/TE technique). Minimally invasive PIP joint arthrodesis with LCP demonstrated low postoperative infection rates. Gross and histological evaluations revealed considerable destruction of the articular cartilage in the LCP/SurD/TE-treated joints. In contrast, almost no destruction of the cartilage was observed in the LCP-treated joints. Our results suggest that the LCP technique alone is not sufficient for PIP joint arthrodesis and that the LCP/SurD/TE technique may be useful for PIP joint arthrodesis in horses.     

in vitro metabolism of azinphosmethyl in susceptible and resistant houseflies

The in vitro metabolism of [¹⁴C-methoxy] or [³²P]azinphosmethyl by subcellular fractions of abdomens from a resistant and a susceptible strain of houseflies was studied. The degradative activity in both strains was associated with the microsomal and soluble fractions and required NADPH and glutathione, respectively. The resistant strain possessed higher activity for both the mixed-function oxidases and the glutathione transferase than the susceptible strain, and both systems appear to be important in the resistance mechanism. The mixed-function oxidases were involved in the oxidative desulfuration as well as the dearylation of azinphosmethyl. A glutathione transferase located in the soluble fraction catalyzed the formation of desmethyl azinphosmethyl and methyl glutathione. This enzyme also demethylated azinphosmethyl oxygen analog. Although the soluble fraction exhibited both glutathione S-alkyltransferase and S-aryltransferase activity against noninsecticidal substrates, no evidence of the transfer of the benzazimide moiety from azinphosmethyl to glutathione was obtained. Sephadex G-100 chromatography of the soluble enzymes revealed a common eluting fraction responsible for both types of transferase activity.     

Reference values of hematological and biochemical parameters for the world smallest microminipigs

In this study, we demonstrated growth curves and reference values for hematological and serum biochemical parameters of Microminipigs, the world smallest experimental minipigs. In both male and female animals, the body weights (BWs) at 3 and 6 months of age were <5 kg and <10 kg, respectively, and growth curve revealed almost plateau (approximately 20 kg BW) after 18 months of age. Major hematological and serum biochemical parameters showed no gender differences and the values were very similar to those in G?ttingen and Yukatan minipigs. The values obtained in this study can serve as fundamental reference, and thereby facilitate the use of Microminipig in life science research.     

Manipulation of tannins in oaks by galling cynipids

We measured the protein precipitation capacity of tannins in the tissues of Quercus crispula and Quercus serrata galls induced by a cynipid (Trigonaspis sp.) to examine the nutrition hypothesis on gall induction. The protein precipitation capacity was significantly lower in the nutritive tissues of galls, on which the cynipids feed, than in ungalled sound leaves and in the outer gall tissues. In addition, the protein precipitation capacities in the leaves and in the outer gall tissues were significantly larger in Q. crispula than in Q. serrata samples, whereas that of the nutritive tissues did not differ between these oak species. These results suggest that gall induction is one of the adaptations of galling cynipids to host–plant defensive compounds, and support the nutrition hypothesis.     

Lateral female strobili production in a Japanese red pine (Pinus densiflora Sieb. et Zucc.) clone by exogenous cytokinin application

Application ofN ⁶-benzylaminopurine (BAP) and gibberellin (GA₃) to a grafted Japanese red pine (Pinus densiflora Sieb. et Zucc.) clone promoted flowering. BAP (250 mg/l) was sprayed five times to the top of branches in July or September 1992. GA₃ (100 or 500 mg/l) was applied five times to the whole plants by spray treatment during the above period, or GA₃ solution of the same concentration was injected once into a small incision made in the main stem of clone trees at the beginning of July or the middle of September 1992. In May 1993, no apical female strobili were observed at the top of the new shoots in any treatment. However, lateral female and bisexual strobili were produced in the lower part of new shoots by BAP application in September, regardless of GA₃ application. BAP applied in September without GA₃ application was sufficient to produce lateral female and bisexual strobili. In such treatment, production of female and bisexual strobili was 47 and 25, and they occurred in 16 out of 68 new shoots, 11 out of 15 treated branches and 5 out of 5 treated trees. The average number of female and bisexual strobili per new shoot was 1.06. BAP treatment either in July or in September significantly decreased (p<0.05) the production of male strobili. These results indicate that BAP application induces the development of lateral female strobili in Japanese red pine when applied during the period of floral differentiation. A part of this paper was presented at the 105th annual meeting of the Japanese Forestry Society.     

Transepithelial transport of alpha-lipoic acid across human intestinal Caco-2 cell monolayers

Alpha-lipoic acid (LA) is used in dietary supplements or food with antioxidative functions. The mechanism for the intestinal absorption of alpha-lipoic acid was investigated in this study by using human intestinal Caco-2 cell monolayers. LA was rapidly transported across the Caco-2 cell monolayers, this transport being energy-dependent, suggesting transporter-mediated transport to be the mechanism involved. The LA transport was strongly dependent on the pH value, being accelerated in the acidic pH range. Furthermore, such monocarboxylic acids as benzoic acid and medium-chain fatty acids significantly inhibited LA transport, suggesting that a proton-linked monocarboxylic acid transporter (MCT) was involved in the intestinal transport of LA. The conversion of LA to the more antioxidative dihydrolipoic acid was also apparent during the transport process.