人类抗菌肽hepcidin在毕赤酵母中的分泌表达和纯化

报道了一种低分子量的人类抗菌肽hepcidin基因的克隆、表达以及纯化.试验证明在毕赤酵母中能成功分泌有活性的hepcidin,hepcidin在重组菌株中的表达量约为3 mg/L,Western blot显示2.2 kD的重组hepcidin条带,重组hepcidin通过凝胶过滤及反向高效液相色谱纯化,质谱验证,重组蛋白对金黄色葡萄球菌和枯草芽孢杆菌表现出抗菌活性.     

Metabolic signatures altered by in vitro temperature stress in <Emphasis Type="Italic">Ajuga bracteosa</Emphasis> Wall. ex. Benth.

To elucidate how biosynthesis of plant metabolites is affected by temperature, metabolite profiles from in vitro regenerated plants raised under different temperature regimes of 10, 15 °C, 20 °C, 25 °C and 30 °C were obtained using electrospray ionization mass spectrometry (ESI-MS), and principal component analysis (PCA) was carried out to identify key metabolites. Several bin masses were detected by PCA loading scatter plots which separated the samples. In-house bin program selectively manifested the putative known metabolites depending on % total ions count and intensity of selected bins in the plant samples. Total phenolic and flavonoid content were harvested to highest levels (12.9 mg GAE/g DW and 9.3 mg QE/g DW), respectively, at 15 °C. Besides, pinoresinol (lignan), some of the vital amino acids such as serine, methionine, histidine and glutamine were found to be at higher amount in plants raised at 15 °C. Significant phenylpropanoids like cinnamic acid, caffeic acid and quercitol were detected at a higher concentration in plants raised at 15 °C as compared to other treatments. However, phosphoenolpyruvate, and oxalosuccinate (intermediates of the pentose phosphate pathway) were accumulated the most in plants raised at 30 °C and they were detected with lowest values at 10 °C. Glucose and deoxy-xylose 5 phosphate (intermediates of TCA cycle) were found in higher amounts at temperature treatments of 15 and 25 °C, respectively. We conclude that a low-temperature treatment (15 °C) results in a stress-induced accumulation of a variety of pharmacologically important secondary metabolites.     

Separate and combined effects of silicon and selenium on salt tolerance of wheat plants

Soil salinity is the leading global abiotic stress which limits agricultural production with an annual increment of 10%. Therefore; a pot experiment was conducted with the aim to alleviate the salinity effects on wheat seedlings through exogenous application of silicon (Si) and selenium (Se). Treatments included in the study were viz. (Ck) control (no NaCl nor Si and Se added), only salinity (50 mM NaCl), salinity + Si (50 mM NaCl with 40 mM Si), salinity + Se (50 mM NaCl with 40 mM Se) and salinity + Si + Se (50 mM NaCl + 40 mM Si + 40 mM Si). The salt stress impaired the growth (root and shoot dry weight, root: shoot ratio, seedlings biomass), water relations, photosynthetic attributes, transpiration rate and chlorophyll contents of wheat seedlings. Nonetheless, the foliar application of Si and Se alone and in combination improved the growth, water relations, photosynthetic attributes, transpiration rate and chlorophyll contents of wheat seedlings under stressed conditions. Moreover, an increase in antioxidant enzyme activity and accumulation of osmo-protectants (proline, soluble protein and soluble sugar) was noted under stressed conditions, which was more pronounced in wheat seedling which experienced combined application of Si and Se. To conclude that, foliar application of Si alone mitigated the adverse effect of salinity, while the combined application of Si and Se was proved to be even more effective in alleviating the toxic effects of salinity stress on wheat seedlings.     

Effects of osmotic pressure on motility, plasma membrane integrity and viability in fresh and frozen-thawed buffalo spermatozoa

In the first experiment, osmotic pressure of semen and seminal plasma in a semen sample from each of the 20 mature Nili-Ravi buffalo bulls was determined. In the second experiment, effects of osmotic pressure on motility (%), plasma membrane integrity (%) and viability (%) in fresh and frozen-thawed semen samples from each of the seven mature Nili-Ravi buffalo bulls was determined. In the first experiment, seminal plasma was harvested by centrifuging semen at 400 × g for 10 min at 37°C and osmotic pressure was determined using an osmometer. In the second experiment, motility (%) was assessed in fresh and frozen-thawed (37°C for 30 s) semen samples using a phase-contrast microscope (×400). Plasma membrane integrity (%) was determined by mixing 50 μl each of fresh and frozen-thawed semen with 500 μl of solution having an osmotic pressure of 50, 100, 150, 190 or 250 mOsm/l (hypotonic treatments of fructose + sodium citrate) and incubating at 37°C for 1 h. Viability (%) of fresh and frozen-thawed spermatozoa before and after challenging them to osmotic pressure (hypotonic treatments) was assessed using supravital stain under a phase-contrast microscope (×400). In the first experiment, the mean ± s.e. osmotic pressures of the buffalo semen and seminal plasma were 268.8 ± 1.17 and 256.0 ± 1.53 mOsm/l, respectively. In the second experiment, motility (%) decreased (P < 0.05) in frozen-thawed semen samples as compared with fresh semen (60.1 ± 1.34 v. 81 ± 1.57, respectively). The plasma membrane integrity (%) and magnitude of osmotic stress in fresh and frozen-thawed semen samples was higher (P < 0.05) at 50, 100, 150 and 190 mOsm/l as compared with 250 mOsm/l. Loss of viability (%) in fresh and frozen-thawed semen samples was higher (P < 0.05) at 50 mOsm/l (59% in fresh, 70% frozen thawed) as compared with other osmotic pressures, while it was lowest at 250 mOsm/l (4.1% for fresh, 9.7% frozen thawed). In conclusion, osmotic pressure of Nili-Ravi buffalo semen and seminal plasma is determined. Furthermore, variation in osmotic pressure below 250 mOsm/l is not favorable to fresh and frozen-thawed buffalo spermatozoa.     

Butyrylcholinesterase, lipoxygenase inhibiting and antifungal alkaloids from Isatis tinctoria

Phytochemical investigations on the alkaloidal fraction of the whole plant of the Isatis tinctoria led to the isolation of the alkaloids 1-6. Compounds 3, 2 were found to be potent butyrylcholinesterase and lipoxygenase enzymes inhibitors in a concentration-dependent manner with the IC(50) values 16.3 +/- 0.06 and 19.7 +/- 0.03 microM against BChE and 30.6 +/- 0.02 and 33.7 +/- 0.05 microM against LOX, respectively. The compounds (1-6) showed significant antifungal activity against Trichophyton schoen leinii, Aspergillus niger, Candida albicans, Trichophyton simii, and Macrophomina phaseolina.     

Endocytosis of hepatitis B immune globulin into hepatocytes inhibits the secretion of hepatitis B virus surface antigen and virions

Hepatitis B immunoglobulin is used for prophylaxis against hepatitis B virus (HBV) and is thought to act by neutralization of virions and hepatitis B virus surface antigen (HBsAg)-containing particles in circulation. Using a panel of hepatocyte-derived cell lines, the present study investigated in vitro whether HBs-specific immunoglobulin G (IgG) is internalized in hepatocytes and whether it interacts with HBsAg in the cells. By immunoelectron microscopy and immunoblotting, human IgG and FcRn receptor for IgG were demonstrated on cellular membranes and in cytoplasmic extracts, irrespective of the HBsAg status of the cells. Furthermore, HBsAg and anti-HBs were shown to be colocalized in the same cellular compartment by two-color confocal microscopy. Endocytosis of HBs-specific IgG caused intracellular accumulation of HBsAg in a dose-dependent manner and inhibited the secretion of HBsAg and HBV virions from the cells. These effects were not observed with F(ab)(2) fragments or nonimmune IgG as controls. The specificity of intracellular HBsAg- anti-HBs interaction was further investigated in cells transfected with HBV genomes expressing wild-type HBsAg or immune escape HBsAg (with a G145R mutation). Monoclonal anti-HBs markedly reduced the secretion of wild-type HBsAg, while the secretion of mutant HBsAg was not affected. These results suggest that HBs-specific IgG binds to hepatocytes and interacts with HBsAg within the cells. This may be relevant for the selection of surface antibody escape mutations.     

Ethnomedicinal flora of Frontier Region Tank,Fata, Pakistan

Main objective of the study was to record the ethnobotanical uses of indigenous plants of the federally administered tribal area (FATA), Bhittani (Local Tribe). Total interviewed local informants of different ages through questionnaire were 212 (196 male and 16 female). Well-known statistical indices, Use Value (UVi) formula and Fidelity Level (FL%) were used for quantification of the recorded data. After identification, the collected specimens were deposited in the Herbarium of Botany Department, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan. Present findings reveal that 38 plants species belonging to 26 families were being used in the treatment of 31 different diseases. Regarding plant habit, herbs were the leading growth form (63%), followed by trees (24%) and shrubs (13%) respectively. Solanaceae (13.16%) was leading family used in curing various diseases, followed by Asteraceae and Moraceae (7.89% each). Most cited families by the informants were Solanaceae (5 species, 137 citations), Asteraceae (3 species, 81 citations), Moraceae (3 species, 69 citations) and Amaryllidaceae (2 species, 57 citations). Most of taxa were used in curing constipation and stomach problem (9.88% each), followed by pain and cough (7.41% each), digestive and fever (4.96% each). Most frequently used plant part for curing different diseases was leaves (33.33%), followed by fruits (21.67%) and roots (13.33%). Medicinal plants with most use values and high ranks were Withania coagulans (0.88) ranked 1^st and Cichorium intybus 2^nd (0.81) while Cichorium intybus also showed 100% FL value. Plants of study area provide most of the basic requirements for the survival of local communities. There is huge pressure exerted on the natural vegetation due to their overuse by the inhabitants of the area. If proper remedial measures are not taken soon, this process may lead to decline of the valuable plant species from the study area.     

Biochar Application Improves the Drought Tolerance in Maize Seedlings

Application of biochar to agricultural soils is mostly used to improve soil fertility. Experimental treatments were comprised of two factors: i) drought at two level, i.e., 80% and 40% water holding capacity (WHC) which was maintained on gravimetric basis ii) three levels of biochar i.e., control, 2 t ha^-1 and 4 t ha^-1 added to soil. Experimentation was done to examine potential of biochar application to enhance the growth attributes, water relations, photosynthetic pigments and antioxidants activities in maize (Zea mays L.) seedlings. Results of study revealed that biochar application increased the growth qualities (total seedlings biomass, dry weight of shoot and root, shoot length and root length). In addition; contents of photosynthetic pigments (chlorophyll a, b, a + b and a/b), water relation (relative water contents, turgor potential, osmotic potential and water potential) were improved significantly due to addition of biochar. Addition of 4 t ha^-1 biochar led to significant rise activity of enzymatic antioxidant catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD) in leaf of maize seedling sunder drought as well as well watered circumstances. However, biochar applied at the rate 4 t ha^-1 improved the all the physiological and biochemical attributes in maize seedlings under drought. From the results it was concluded that biochar application is an efficient way to alleviate adverse effect of drought stress on maize. In drought prone areas, long term impacts of biochar on production of maize and properties of soil could be recommended as upcoming shove.     

Differentially methylated genomic fragments related with sexual dimorphism of rice pests,Sogatella furcifera

Sogatella furcifera (Hovarth) is a major rice pest with sexual dimorphism. The objective of the current research was to monitor differentially cytosine methylation at CCGG sequences in male and female adults of S. furcifera to determine the association between gene methylation and sexual phenotypes using methylation‐sensitive representational difference analysis. After the second subtractive hybridization, four differentially methylated DNA bands were obtained and sequenced. Ten different fragments were found. One fragment from the positive hybridization was 120 bp, and highly similar to the tramtrack genes from Nasonia vitripennis. Another fragment from the reverse hybridization was 414 bp, and homologous to the 28S rRNA gene of S. furcifera with a similarity rate as high as 99%. We also discussed how DNA methylation of tramtrack and 28S rRNA genes produced effects on sexual differentiation and development. These results provide potential evidence that DNA methylation of some genes may be related to sexual phenotype variations in S. furcifera and will facilitate future studies on the epigenetic mechanisms of insect sexual dimorphism.