Diverse functions of perlecan in central nervous system cells in vitro

Therapeutic treatment targeting one cell type is considered ineffective in remedying any injury to the central nervous system (CNS). Perlecan, a multi‐functional, heparan sulfate proteoglycan, shows diverse effects on distinct cell types, suggesting that it is one of the candidates that can augment the regenerative mechanisms in the injured CNS. Therefore, we examined the functions of perlecan in CNS cells in vitro by using perlecan purified from bovine kidney. Perlecan‐coated cell culture plates, unlike their type I/III collagen‐coated counterparts, did not inhibit the adhesion of neural stem/progenitor cells (NS/PCs) and neurons. The coated perlecan and the perlecan added to the culture medium suppressed astrocyte proliferation; however, perlecan added to the medium promoted NS/PC proliferation. Neurons were promoted to extend their neurites on the perlecan‐coated substrate, and perlecan added to the medium also showed a similar effect. NS/PC proliferation and neurite extension is a major regenerative reaction in CNS injury, whereas excess proliferation of astrocytes cause hypertrophy of glial scars, which repels neurons. Our in vitro study suggests that perlecan is an attractive candidate to promote regenerative mechanisms and to suppress reactions that hamper regenerative processes in cases of CNS injury.     

Disruption of endogenous perlecan function improves differentiation of rat articular chondrocytes in vitro

Heparan sulfate (HS) and heparan sulfate proteoglycans (HSPG) are necessary for normal cartilage development and chondrocyte differentiation. However, recent studies demonstrated that HSPG accelerate dedifferentiation and catabolism in chondrocytes from degenerative cartilage. In this study, we investigated the inhibitory effect of HSPG on chondrocyte differentiation in vitro. Rat articular chondrocytes were cultured at low (0.3 × 10⁴ cells/cm²) and high (1.5 × 10⁵ cells/cm²) density in the presence or absence of heparitinase I, an HS degrading enzyme. Cells cultured at low density dedifferentiated and exhibited an elongated morphology, and treatment with heparitinase I precluded cell elongation. Conversely, populations of chondrocytes cultured at high density exhibited either a dedifferentiated or differentiated phenotype. Glycosaminoglycan accumulation increased in heparitinase I‐treated cells. To determine the function of perlecan, an important HSPG for cartilage development, in chondrocyte differentiation, rat chondrocyte cultures were exposed to an anti‐perlecan antiserum to inhibit perlecan function. Western blotting analysis indicated that preventing perlecan activity increased type II collagen synthesis. Our results suggest that HSPG are negative regulators of chondrocyte differentiation in vitro and that perlecan contributes to chondrocyte dedifferentiation in vitro.     

Simple method of genomic DNA extraction from Rhizobia in dried nodules of Phaseolus vulgaris for strain differentiation by PCR-based DNA fingerprinting

Repetitive DNA peR fingerprinting of bacterial genomic DNA is a useful tool for typing and differentiation of rhizobial strains. The method was reported to be suitable for strain differentiation of Rhizobia present in individual root nodules of some leguminous plants without the need for isolation and cultivation of the strains, in which rhizobial genomic DNA was extracted directly from each fresh or frozen nodule. We developed a new protocol of rhizobial genomic DNA extraction/purification from dried nodules of Phaseolus vulgaris for generating repetitive DNA peR fingerprints of Rhizobia present in the nodules. The simplified protocol consists of only three major steps, heat extraction of genomic DNA from rhizobial cells prepared from dried nodules, ethanol precipitation of the DNA and Sephadex G-50 column purification of the DNA, and generated fingerprints with good quality for differentiation of Rhizobia strains. The protocol will be useful to examine the nodule occupancy of inoculated rhizobial strains in field experiments.     

Improved survival and growth performances with photoperiod and feeding schedule manipulation in bagrid catfish Mystus nemurus (Cuvier & Valenciennes 1840) larvae

Manipulation of photoperiod: 24 h light (24L), 12 h light:12 h dark (12L:12D) and 24 h dark (24D); and feeding schedules: day and night feeding (DNF), day feeding (DF) and night feeding (NF) was conducted to determine effects on survival, cannibalism and growth of larval bagrid catfish Mystus nemurus 2–14 days after hatching (dAH). Photoperiod insignificantly affected all parameters. Feeding schedule significantly affected survival and total length at 6 (P < 0.049; P < 0.009), 10 (P < 0.033; P < 0.000) and 14 dAH (P < 0.013; P < 0.000), respectively, but affected cannibalism at 10 (P < 0.043) and 14 dAH (P < 0.013). Survival for DNF was significantly higher than DF. Cannibalism for DNF was significantly lower than NF at 10 and 14 dAH. Total length for DNF was significantly higher than DF and NF at 10 and 14 dAH. At 14 dAH, feeding schedule significantly affected feed intake, final weight and coefficient of variation. For feed intake and final weight, DNF was significantly higher than DF and NF. For coefficient of variation, NF was significantly higher than DF. This study suggests that larval bagrid catfish can be reared at 24L, 12L:12D or 24D but should be fed day and night for improved growth, survival and reduced cannibalism.     

Comparison on growth performance,body coloration changes and stress response of juvenile river catfish,Pangasius hypophthalmus reared in different tank background colour

The effects of different tank background colours (green, black and white) on the growth performance and body coloration of juvenile river cafish, Pangasius hypophthalmus, were examined in this study. The experimental fish with an initial body weight (BW) 0.13 ± 0.05 g and total length (TL) 2.54 ± 0.37 cm were used, and groups of 15 fish per tank were randomly distributed. The experiment was conducted for 20 days as a triplicate for each treatment. Survival, growth in BW, body weight gain (BWG), total length (TL), total length gain (TLG), specific growth rate (SGR), condition factor, feed conversion ratio (FCR), swimming speed, body coloration and stress response were evaluated. River catfish reared in white and green tanks attained significantly higher BW, BWG, TL, TLG and SGR compared to the black tank. The white tank attained significantly better FCR compared to the black tank. River catfish showed the most active swimming behaviour in the green tank (5.49 ± 1.32 cm/s) while the least in the white tank (3.37 ± 0.93 cm/s). Body coloration of river catfish in the black tank showed almost pure black body coloration (75.59 ± 9.08), followed by the green tank (126.23 ± 17.46) and the white tank (148.19 ± 10.51). However, there was no significant difference in the stress level of river catfish reared in white and black tanks and insignificant to those reared in the green tank. The findings in this study showed that the white and green background colour tanks can be used for promoting growth of river catfish culture while the black background colour tank can enhance body coloration that is suitable as ornamental fish.     

Optimum low salinity to reduce cannibalism and improve survival of the larvae of freshwater African catfish <Emphasis Type="Italic">Clarias gariepinus</Emphasis>

The freshwater African catfish Clarias gariepinus is carnivorous and cannibalistic even during the larval and juvenile stages and this behavior causes economic losses in aquaculture. This study examined for the first time the effect of salinity on cannibalism, survival, and growth of African catfish larvae in the hatchery. Larvae (4 days old, median 7.8 mm TL, 2.8 mg BW) of the African catfish were reared for 21 days at nominal salinity 0, 1, 2, 3, 4, 5, 6, and 7 ppt. After 21 days, they grew to 10–39 mm (median 22 mm) and 10–490 mg (median 90 mg), with no significant difference by salinity treatments. Survival ratios were similarly low (24–31%) at 0, 1, 3, and 7 ppt and significantly higher (49–55%) at 2, 4, 5, and 6 ppt. Cannibalism was significantly lower, 15–30% at 4–6 ppt, than the 40–50% at 0–3 and 7 ppt. Size variation was lower at 4–6 ppt and higher at 0–3 and 7 ppt. We recommend hatchery rearing of African catfish at the optimum low salinity of 4–6 ppt rather than in full fresh water at least up to 21 days. This rearing method fosters larval welfare and improves hatchery production.