In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig

In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.

Influence of green manures on epiedaphic macrofauna in cerrado/pantanal ecotone / Influência de adubos verdes sobre a macrofauna epiedáfica no ecótono cerrado/pantanal

The aim of this study was to determine the similarity between nine species of green manure regarding the occurrence of epiedaphic macrofauna. The experiment was conducted during the months of August 2014 to July 2015 at the Crop Science sector of the State University of Mato Grosso do Sul, Unit of Aquidauana. Treatmentes evaluated were: Canavalia ensiformes, Dolichos lablab, Mucuna pruriens, Mucuna aterrima, Crotalaria spectabilis, Crotalaria juncea, Crotalaria ochroleuca, Cajanus cajan, Cajanus cajan and two control (fallow area and native vegetation). Experimental design was completely randomized with two replications. Each plot consists of 10 lines with 4.5 m long. Epiedaphic macrofauna inhabiting the interphase between the vegetation cover and the soil surface was collected weekly by the Pitfall model. Individuals collected in all green manure species in the same month were added and subjected to cluster analysis. The months of September and October composed group 2 due to higher averages of epiedaphic macrofauna individuals. The month of July had the highest number of individuals from Isoptera order, composing the group 3. The month of November was in which was collected the greatest number of individuals from Hymenoptera order, which has allocated this month in group 4. The others months of the year formed the group 1, indicating that the region climatic conditions in this period do not affect significantly the distribution of epiedaphic macrofauna individuals.

O objetivo deste trabalho foi verificar a similaridade entre nove espécies de adubo verde quanto à ocorrência de macrofauna epiedáfica. O experimento foi conduzido durante os meses de agosto de 2014 a julho de 2015 no setor de Fitotecnia da Universidade Estadual de Mato Grosso do Sul, Unidade de Aquidauana. Os tratamentos avaliados foram: Canavalia ensiformes, Dolichos lablab, Mucuna pruriens, Mucuna aterrima, Crotalaria spectabilis, Crotalaria juncea, Crotalaria ochroleuca, Cajanus cajan, Cajanus cajan e duas testemunhas (área de pousio e mata nativa). O delineamento experimental utilizado foi o inteiramente casualizado, com duas repetições. Cada parcela foi composta por 10 linhas com 4,5 m de comprimento. A macrofauna epiedáfica, que habita na interfase entre a cobertura vegetal e a superfície do solo foi coletada semanalmente por meio de armadilhas modelo Pitfall (de queda). Os indivíduos coletados em todas as espécies de adubação verde no mesmo mês foram somados e submetidos à análise de agrupamento. Os meses de setembro e outubro compuseram o grupo 2 devido as maiores médias de indivíduos da macrofauna epiedáfica. O mês de julho obteve o maior número de indivíduos da ordem Isoptera, compondo o grupo 3. O mês de novembro foi no qual foi coletado o maior número de indivíduos da ordem Hymenoptera, o que alocou este mês no grupo 4. Os demais meses do ano formaram o grupo 1, o que indica que as condições climáticas da região neste período não afetam significativamente a distribuição dos indivíduos da macrofauna epiedáfica.

Purification of a protease inhibitor from Dolichos biflorus using immobilized metal affinity chromatography.

Plant protease inhibitors (PIs) are generally small proteins which play key roles in regulation of endogenous proteases and may exhibit antifeedant, antifungal, antitumor and cytokine inducing activities. Dolichos biflorus (horse gram) is an unexploited legume, which is rich in nutrients and also has therapeutic importance. It contains a double-headed PI, which is an anti-nutritional factor. As there is no report available on its simultaneous removal and purification in single step, in this study, a double-headed PI active against both trypsin and chymotrypsin was purified from Dolichos biflorus to ~14-fold with ~84% recovery using an immobilized metal affinity chromatography (IMAC) medium consisting of Zn-alginate beads. The method was single-step, fast, simple, reliable and economical. The purified inhibitor showed a single band on SDS-PAGE corresponding to molecular mass of 16 kDa and was stable over a pH range of 2.0-12.0 and up to a temperature of 100°C for 20 min. The optimum temperature for trypsin and chymotrypsin inhibitor was observed to be 50°C and 37°C, respectively and pH optimum was pH 7.0 and 8.0, respectively. Thus, IMAC using Zn-alginate beads was useful in simultaneous purification and removal of an anti-nutritional factor from horse gram flour in single step. This procedure may also be employed for purification of other plant PIs in one step.

A morphological study of vomeronasal organ of Korean black goat (Capra aegagrus hircus) / 대한수의학회지

The vomeronasal organ (VNO) plays an important role in reproduction and social activities in ruminants including goats. A morphological study on the structure of VNO and its epithelial cells was carried out in Korean black goats. Grossly, the VNO of Korean goats opens into mouth through incisive ducts. Microscopically, the epithelium of VNO consisted of medial sensory epithelium and lateral non-sensory epithelium. Several blood vessels and nerve bundles were observed in the lamina propria encased by vomeronasal cartilage. Immunohistochemical staining showed that protein gene product (PGP) 9.5 was immunostained in the receptor cells of the sensory epithelium and in some cells of the non-sensory epithelium. Galectin-3 was mainly observed in the supporting cells of sensory and non-sensory epithelium. Lectins including wheat germ agglutinin, Ulex europaeus agglutinin, Bandeiraea simplicifolia lectin Isolectin B4, Dolichos biflorus agglutinin and soybean agglutinin used in this study were bound in VNO sensory, non-sensory epithelia as well as in the lamina propria with varying intensity. Collectively, this is a first descriptive morphological study of VNO of Korean black goat with special reference to lectin histochemistry.

Histochemical study of lectin-binding patterns in the rat vomeronasal organ during postnatal development / 대한수의학회지

Histochemical patterns of lectin binding during development of the rat vomeronasal organ (VNO) were studied to determine whether glycoconjugates are differently expressed after birth. Three types of lectins, Dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA), and Ulex europaeus agglutinin I (UEA-I), were studied histochemically in the rat VNO at various stages post-birth: postnatal days 1 and 7, the preweaning period (4 weeks after birth), and at sexual maturity (8 weeks after birth). The free border of the vomeronasal sensory epithelium was positive for both WGA and UEA-I in rats of all ages; whereas, VNO receptor cells and supporting cells were positive only for both WGA and UEA-I from 4 weeks after birth. DBA reactivity was detected in the free border but less so in receptor cells and supporting cells. WGA and UEA-I, but not DBA, showed similar patterns in various ages. In the Jacobson's gland, WGA, UEA-I and DBA were detected in some acini from 4 weeks after birth but not at postnatal days 1 or 7. Collectively, reactivity for three lectins, WGA, UEA-I and DBA, increased in receptor cells and gland acini during postnatal development, possibly contributing to the enhanced chemoreception in rats.

Desenvolvimento de plantas de cobertura em solo compactado / Cover crops growth in compacted soil

O objetivo deste trabalho foi avaliar o crescimento aéreo e radicular de três espécies de plantas de cobertura em condições de solo compactado. O experimento foi realizado em vasos montados com anéis de cano PVC. Cada vaso foi constituído por três anéis, sendo os anéis das extremidades com 15 cm de altura e o anel intermediário com 5 cm de altura, todos contendo 20cm de diâmetro. No anel intermediário acomodou-se solo de forma que o mesmo apresenta-se densidades de 1,6 Kg dm-3 (compactado) e de 1,1 Kg dm-3 (não compactado). Após a montagem, os vasos foram cultivados com as espécies de plantas de cobertura: labe labe (Dolichos lab lab), sorgo (Sorghum bicolor L.) e braquiária (Brachiaria ruziziensis). O experimento foi conduzido delineamento inteiramente casualizado com quatro repetições, em esquema fatorial 3x2 (três espécies e dois níveis de compactação. Após 54 dias da emergência das plantas determinou-se a massa seca da parte aérea e a massa seca das raízes em cada camada dos vasos. A compactação do solo não reduziu produção de massa seca da parte aérea e radicular das espécies. Na presença de solo compactado, houve aumento no crescimento radicular de Brachiaria ruziziensis na camada intermediária do vaso. O sorgo e a braquiária foram as espécies com maior potencial para estruturar solos compactados. O sorgo foi a espécie com maior eficiência para colonização de raízes na camada inferior do vaso, independente da presença ou não de camada intermediária compactada.

The aim of this study was to evaluate shoot and root development of three cover crop species in compacted soil. The experiment was conducted in pots fitted with rings of PVC pipe with 20 cm of internal diameter. Each pot was constituted by three rings. The extremities ring had 15 cm high and middle ring had 5 cm. At the intermediate ring the bulk density for 1.6 Kg dm-3 (compressed) or 1.1 Kg dm-3 (uncompressed) was adjusted. The pots were cultivated with the following cover crops: labe labe (Dolichos lab lab), sorgo (Sorghum bicolor L.) and Brachiaria ruziziensis. The study was carried out in a in a completely randomized design, with four replicates, in a 3x2 factorial arrangement (three species and two compaction levels). After 54 days of plant emergence, shoot and root dry matter were evaluated in each layer of the pots. The treatments were analyzed by analysis of variance and means compared by Tukey test at 5%. Soil compaction did not reduce shoot and root dry matter of cover crops.There was an increase in root growth of Brachiaria ruziziensis in the middle layer of the plot, in compacted soil. Sorghum and pasture were the species with the greatest potential for structuring compacted soil. Sorghum was the species with greater efficiency for root colonization in the lower layer of the plot, regardless of the presence or absence of middle layer compacted.

Hypolipidemic effect of methanolic extract of Dolichos biflorus Linn. in high fat diet fed rats.

High fat diet fed rats showed significant increased levels of plasma and tissue total cholesterol, triglycerides, free fatty acids, phospholipids, plasma LDL cholesterol and decreased level of plasma HDL cholesterol. Methanolic extract of D. biflorus administration to high fat diet fed rats showed near to normal levels of the above lipids in plasma and tissues. Higher dose of the extract (400 mg/kg body weight) showed comparable results with standard drug atorvastatin. It is concluded that the methanolic extract of D. biflorus possesses hypolipidemic activity in high fat diet fed rats.

Effect of Cr in irrigation water on germination and growth of French beans (Dolichos lablab L.).

The waters containing chromium (Cr) mainly from industries enter into the sewers, municipal treatment plant, streams and lakes that are used for irrigation purposes. The effect of Cr in irrigation water on seed germination and growth of French bean (Dolichos lablab L.) was investigated. Germination studies were conducted in the lab and field conditions. Seeds were treated with different concentrations (0 to 25 mg/1) of Cr in irrigation water. The percentage germination of seeds and plant growth showed a gradual decrease with increase in Cr concentration. Stimulatory effect in growth of plant was observed at lower concentration (1 to 2 mg/l). Accumulation of Cr was in the order of Root > Leaf > Stem > Pod. There was an increase in the total concentration of Cr content of soil with increase in Cr concentration in irrigation water.

Lectins Binding in Squamous Cell Carcinoma of the Uterine Cervix as a Diagnostic and Prognostic Marker / 대한산부인과학회잡지

OBJECTIVE: The aim of the study was to evaluate the corelation between an expression of lectins and survival of patients with squamous cell carcinoma of uterine cervix. METHODS: The cell surface carbohydrate profile of formalin-fixed paraffin embedded tissue section of squamous cell carcinoma of the uterine cervix was evaluated using ulex europaeus agglutinin(UEA-1), peanut agglutinin(PNA), dolichos biflorus agglutinin(DBA), soybean agglutinin(SBA) and lotus tetragonobus lectin(LTL) by the avidin-biotin complex method. Fifty-one cases of squamous cell carcinoma and 10 cases of normal squamous epithelium of the uterine cervix were selected from the file which were treated during Jan. 89 to 31 Dec. 1992 in Department of Obstetrics and Gynecology, Korea University Hospital. RESULTS: UEA-1 and LTL were negative in normal cervix while positive in 76.5%, 47.1% of squamous cell carcinoma respectively, and useful markers for differential diagnosis between normal and squamous cell carcinoma. SBA and LTL were useful for differential diagnosis of keratinizing and non-keratinizing squamous cell carcinoma. UEA-1 may play an important role in lymphovascular invasion of squamous cell carcinoma of the uterine cervix. CONCLUSIONS: There was no correlation among clinical staging, patient's survival and lectins binding in squamous cell carcinoma.

The significance of lectins in the differentiation between lichen sclerosus et atrophicus(LSA)and scleroderma / 대한피부과학회지

BACKGROUND: Differential diagnosis of lichen sclerosus et atrophicus( SA) and scleroderma is occasionally difficult. OBJECTIVE: The purpose of this study was to attempt differentiation between the two diseases using imrnunohistochemical stain and lectins. MEHTODS: Paraffin-embeddred sections of 4 cases of LSA and 11 cases of scleroderma were evaluated for this study. Using lectins, such as peanut agglutinin(PNA), siybean agglutinin(SBA), Ulex europaeus agglutinin-I(UEA-I) and Dolichos biflorus agglutinin(DBA) and the avidin-biotin-peroxi-dase complex(ABC) technique, differential lectin binding patterns betv een the two diseases were examined. RESULTS: In the case of LSA, PNA and SBA stained the upper and lower spinous layer of the epidermis, and UEA I also stained the spinous layer of the epidermis weakly, but no DBA was stained. In the case of scleroderma, PNA stained not only the spinous layer but also the basal layer, SBA stained the upper half of the spinous layer but not the lower half of the pinous layer of epidermis. But UEA-I stained the vascular endothelial cells of dermis instead of epidermis, and DBA stained only the basal layer of epidermis. CONCLUSION: Staining of these 4 lectins on paraffin-embedded sectians using ABC teehnique could be helpful in differenting LSA and scleroderma.