Strain improvement,optimization and purification studies for enhanced production of streptokinase from <Emphasis Type="Italic">Streptococcus uberis</Emphasis> TNA-M1

Background

Screening of isolates for their potency to produce streptokinase was an important criterion of this research. The current study emphasizes the strain improvement, optimization and purification studies for enhanced production of streptokinase from Streptococcus uberis TNA-M1 isolated from bovine milk.

Methods

The study was carried out on samples collected from milk sample. Primary screening and characterization is used as an excellent source for the isolation of β-hemolytic organisms. Strain improvement was done by both physical & chemical mutagenesis. The enzyme activity was checked by clot lysis assay and confirmed by fibrin plate method. The partially purified and crude enzyme were analysed by high-performance liquid chromatography. Molecular weight & enzyme purity was checked by SDS–PAGE, further confirmed by fibrin zymography.

Results

Out of the 3 isolated strains, only one isolate expressed β-haemolysis with streptokinase (SK) activity. Based on the results of radial caseinolytic assay and blood clot dissolving assay, isolate TNA-M1 demonstrated the highest streptokinase activity. Based on morphological, biochemical and molecular characterization, it was identified as Streptococcus uberis and the strain was named as Streptococcus uberis TNA-M1. The results indicated that ultra-violet (UV) and ethyl methane sulfonate (EMS) were effective mutagenic agents for strain improvement of Streptococcus uberis TNA-M1 and enhanced SK productivity. HPLC analysis was performed in order to confirm the presence of streptokinase with the similar retention time (0.875 min) with its standard (0.854) min. SDS-PAGE of the enzyme showed protein band of approximately 47 kDa and confirmed by fibrin zymography. It exhibited fibrinolytic activity, which was more potent than other fibrinolytic enzymes. Glucose and peptone were recorded to be the optimum carbon and nitrogen sources respectively.

Conclusion

Thus this study presents its novelty by highlighting the potential of Streptococcus uberis TNA-M1 as a significant source for the production of fibrinolytic enzymes.

     

Stress‐induced changes in color expression mediated by iridophores in a polymorphic lizard

Stress is an important potential factor mediating a broad range of cellular pathways, including those involved in condition‐dependent (i.e., honest) color signal expression. However, the cellular mechanisms underlying the relationship between stress and color expression are largely unknown. We artificially elevated circulating corticosterone levels in male tawny dragon lizards, Ctenophorus decresii, to assess the effect of stress on the throat color signal. Corticosterone treatment increased luminance (paler throat coloration) and decreased the proportion of gray, thereby influencing the gray reticulations that produce unique patterning. The magnitude of change in luminance for corticosterone‐treated individuals in our study was around 6 “just noticeable differences” to the tawny dragon visual system, suggesting that lizards are likely to be able to perceive the measured variation. Transmission electron microscopy (TEM) of iridophore cells indicated that luminance increased with increasing density of iridophore cells and increased spacing (and/or reduced size) of crystalline guanine platelets within them. Crystal spacing within iridophores also differed between skin colors, being greater in cream than either gray or yellow skin and greater in orange than yellow skin. Our results demonstrate that stress detectably impacts signal expression (luminance and patterning), which may provide information on individual condition. This effect is likely to be mediated, at least in part, by structural coloration produced by iridophore cells.     

Bis- and mono-substituted Chalcones exert anti-feedant and toxic effects on fall armyworm Spodoptera frugiperda

Spodoptera frugiperda is a highly polyphagous migratory lepidopteran pest species. It causes infestation in crops leading to the severe crop losses. Being a new invasive parasite, its susceptibility to insecticides needs to be explored; and therefore, there is an urgent need to develop the potent insecticides for the effective control of this insect pest. To attain the crop sustainability, the antifeedant, toxicity and nutritional effects on larvae of Spodoptera frugiperda were studied with six mono- and eight bis- substituted chalcones. The antifeedant activity was calculated when 50% of the larvae control ate 50% of the diet through the FR factor. Toxicity was assessed through larval, pupal mortality and the emergence of adults and nutritional effects with consumption rates (IC), growth (GR) and consumption efficiency (EIC). The bis-chalcones 6b, 6e, 6f and 6h caused lethal effect on S. frugiperda in the first larval stages, being 6b the most toxic (85%). Adults who survived showed malformations and decreased size, which led to death. The larvae fed with aggregate in the bis-chalcones diet: 6b, 6e and 6f had the highest percentage of intake and the poorest conversion of nutrient absorption (ECI), which suggests that the larva metabolizes food for energy and results in a decrease of growth and death in early stages. Bis-chalcones showed more toxicity than mono-chalcones and 6b causes the most toxic and dietary change.     

Deception and the origin of honest signals

Deceptive signals are a challenge to explain because on average, signals should be reliable. When being deceived is costly to the receiver, a coevolutionary struggle between senders and receivers can ensue. Recent work by Macías Garcia and Ramirez raises the intriguing possibility that through such a coevolutionary process, cheats can become honest.     

Treatment with bindarit, a blocker of MCP-1 synthesis, protects mice against acute pancreatitis

Chemokines are believed to play a key role in the pathogenesis of acute pancreatitis. We have earlier shown that pancreatic acinar cells produce the chemokine monocyte chemotactic protein (MCP)-1 in response to caerulein hyperstimulation, demonstrating that acinar-derived MCP-1 is an early mediator of inflammation in acute pancreatitis. Blocking chemokine production or action is a major target for pharmacological intervention in a variety of inflammatory diseases, such as acute pancreatitis. 2-Methyl-2-[[1-(phenylmethyl)-1H-indazol-3yl]methoxy]propanoic acid (bindarit) has been shown to preferentially inhibit MCP-1 production in vitro in monocytes and in vivo without affecting the production of the cytokines IL-1, IL-6, or the chemokines IL-8, protein macrophage inflammatory-1alpha, and RANTES. The present study aimed to define the role of MCP-1 in acute pancreatitis with the use of bindarit. In a model of acute pancreatitis induced by caerulein hyperstimulation, prophylactic as well as therapeutic treatment with bindarit significantly reduced MCP-1 levels in the pancreas. Also, this treatment significantly protected mice against acute pancreatitis as evident by attenuated hyperamylasemia neutrophil sequestration in the pancreas (pancreatic MPO activity), and pancreatic acinar cell injury/necrosis on histological examination of pancreas sections.     

Sequence analysis of the hypervariable regions of the 56 kDa immunodominant protein genes of Orientia tsutsugamushi strains in Malaysia

The DNA sequences encompassing two hypervariable regions, VD II and III of the 56 kDa immunodominant protein gene of 21 Malaysian strains of Orientia tsutsugamushi were determined. Two strains demonstrated a 100% DNA homology with the Gilliam prototype strain, and one with TH1817 strain and TA678 strain respectively. High percentages of DNA similarity (95-99%) were observed with Karp (4 strains), Gilliam (2 strains), TH1817 (4 strains), TC586 (3 strains) and TA763 (1 strain). The remaining strains demonstrated the highest DNA similarity with TA763 (1 strain, 89%), TA678 (1 strain, 86%) and TA686 (1 strain, 87%). Our study provides additional evidence on the existence and the genetic heterogeneity of TA strains of the Southeast Asia and their closely related strains in Malaysia.     

Extract from Clerodendron colebrookianum Walp protects rat heart against oxidative stress induced by ischemic-reperfusion injury (IRI)

Reactive oxygen species (ROS) have pathogenic effects on ischemic-reperfusion injury of heart. Hence, it is important to identify natural antioxidative agents to mitigate such effects. Recently, it has been reported that Clerodendron colebrookianum (CC) leaf extract has antioxidant and hypolipidemic effects in experimental animals. The aim of this study was to examine whether acute treatment with CC extract offers protection against ischemic-reperfusion injury (IRI) and IRI-induced changes in endogenous antioxidant enzyme activities of rat heart. Isolated rat hearts were perfused using the Langendorff's technique, and 20 min of global ischemia was followed by 40 min of reperfusion. Lipid peroxidation after the ischemic-reperfusion episode was significantly reduced in the CC extract-treated heart compared to the control group and suppressed the leakage of lactate dehydrogenase (LDH) during reperfusion. Moreover, CC extract diminished the depletion of myocardial antioxidant enzymes (SOD, Catalase, GSH and GPx) after ischemia-reperfusion. Furthermore, IRI-induced cellular damage was significantly less in CC extract treated myocytes. These results indicate that CC leaf extract protects against oxidative stress and cellular injury associated with ischemic-reperfusion injury of rat heart and suggests that the protective effects of CC extract depend on its antioxidant properties.     

Sacred groves of Manipur,northeast India:biodiversity value,status and strategies for their conservation

The people of Manipur, a state in northeast India, follow ancestral worship and animism in the form of deity worship, with the central focus on worship in forest patches. The beliefs and taboos associated with the Sylvan deities (Umanglais) in the forest patches are restricted to any sort of disturbance of flora and fauna. These social boundaries help to conserve the entire organism as a whole, which stand the concept of sacred groves. The pleasing of deities is performed every year by the Meiteis, a dominant community of Manipur, in honour of the deities and to gain their favour. Indigenous cultural and rituals practices of the local people in sacred groves serve as a tool for conserving biodiversity. Sacred groves are distributed over a wide ecosystem and help in conservation of rare and endemic species. Well-preserved sacred groves are store houses of valuable medicinal and other plants having high economic value, and serve as a refuge to threatened species. One hundred and sixty-six sacred groves were inventoried in Manipur valley that comprises Imphal east, Imphal west, Thoubal and Bishnupur districts of the state. Detailed studies were carried out in four selected sacred groves, to know the importance of biodiversity status and vegetation characteristics. A total of 173 plant species representing 145 genera under 70 families were recorded through baseline floristic survey. The species diversity indices were compared among the four studied groves. The vegetation composition and community characteristics were recorded. Ethnobotanical uses of species were examined, which reveal that 96% of the species were used as medicine for the treatment of various ailments. Utilization of herbal medicine by the Meiteis is closely related to the cultural and ritual practices. A few of the medicinal plants which have disappeared from the locality are now confined only to the groves. Socio-cultural aspects were investigated taking into account the attitudes of local people, which indicate social beliefs and taboo are eroding, simultaneously degrading the degree of protection of sacred groves. Therefore, conservation measures of sacred groves need to be formulated considering the factor of degradation and the basic necessities of the local people. Until and unless a viable option is provided to the local people (especially those who habitat nearby the adjoining areas) for sustaining their economic condition, no step for conservation of biodiversity will be successful.     

Stem cells in postnatal myogenesis: molecular mechanisms of satellite cell quiescence, activation and replenishment

Satellite cells are the primary stem cells in adult skeletal muscle, and are responsible for postnatal muscle growth, hypertrophy and regeneration. In mature muscle, most satellite cells are in a quiescent state, but they activate and begin proliferating in response to extrinsic signals. Following activation, a subset of satellite cell progeny returns to the quiescent state during the process of self-renewal. Here, we review recent studies of satellite cell biology and focus on the key transitions from the quiescent state to the state of proliferative activation and myogenic lineage progression and back to the quiescent state. The molecular mechanisms of these transitions are considered in the context of the biology of the satellite cell niche, changes with age, and interactions with established pathways of myogenic commitment and differentiation.