Search

High temperature negatively affects cotton production worldwide. In Pakistan, cotton crop faces high temperature at peak flowering during June–July, which is a major reason for yield losses. The present study was conducted to find some agro-physiological markers for high-temperature tolerance. Fifty cotton genotypes were raised under normal sown (high-temperature stress) and late sown (optimum temperature) conditions for 2 years. Data were recorded for relative cell injury percentage (RCI), chlorophyll content (CC), canopy temperature (CT), boll retention percentage (BR), bolls per plant (BP), boll weight (BW), hundred seed weight (HSW), sympodial branches per plant (SBP), plant height (PH) and seed cotton yield (SCY). A large amount of variability was found among cotton genotypes as proved by descriptive statistics. Stable genotypes were selected based on higher SCY and yield components (BP, BW, HSW, SBP, PH and SCY) performance along with less RCI%, high CC and low CT. The results of heatmap analysis showed that the six cotton genotypes viz., FH-Noor, FH-Lalazar, FH-458, FH-466, NIAB-545 and NIAB-878 performed better under high-temperature stress. These selected genotypes can be a source for breeding high-temperature tolerance. Furthermore, the results from correlation analysis confirmed that the traits such as RCI%, CC and CT can be considered as early selection criteria due to their positive association with SCY. Whilst some other parameters such as BP, BW, BR, SBP and PH might be used as direct selection indices for SCY under high-temperature stress.

Toxoplasma gondii and Cryptosporidium parvum are protozoan parasites that are highly prevalent and opportunistically infect humans worldwide, but for which completely effective and safe medications are lacking. Herein, we synthesized a series of novel small molecules bearing the diacyl urea scaffold and related structures, and screened them for in vitro cytotoxicity and antiparasitic activity against T. gondii and C. parvum. We identified one compound (GMG-1-09), and four compounds (JS-1-09, JS-2-20, JS-2-35 and JS-2-49) with efficacy against C. parvum and T. gondii, respectively, at low micromolar concentrations and showed appreciable selectivity in human host cells. Among the four compounds with efficacy against T. gondii, JS-1-09 representing the diacyl urea scaffold was the most effective, with an anti-Toxoplasma IC50 concentration (1.21 μM) that was nearly 53-fold lower than its cytotoxicity IC50 concentration, indicating that this compound has a good selectivity index. The other three compounds (JS-2-20, JS-2-35 and JS-2-49) were structurally more divergent from JS-1-09 as they represent the acyl urea and acyl carbamate scaffold. This appeared to correlate with their anti-Toxoplasma activity, suggesting that these compounds’ potency can likely be enhanced by selective structural modifications. One compound, GMG-1-09 representing acyl carbamate scaffold, depicted in vitro efficacy against C. parvum with an IC50 concentration (32.24 μM) that was 14-fold lower than its cytotoxicity IC50 concentration in a human intestinal cell line. Together, our studies unveil a series of novel synthetic acyl/diacyl urea and acyl carbamate scaffold-based small molecule compounds with micromolar activity against T. gondii and C. parvum that can be explored further for the development of the much-needed novel anti-protozoal drugs., Graphical abstract Image 1, Highlights • A focused library of acyl/diacyl ureas and acyl carbamates was efficiently prepared. • Select compounds inhibited the in vitro growth of T. gondii and C. parvum parasites. • The diacyl urea compound JS-1-09 showed good activity against T. gondii parasites. • The identified hit shows potential for further development as novel antiprotozoal agent.

The herbal use for medical purposes is increasing world over. Aloe vera is onesuch herb with established anti-inflammatory action. It has great prospect in terms of replacingtraditional NSAIDS due to better side effect profile on gastrointestinal tract but its effect onplatelet aggregation is what needs to be determined. Objectives: This study was designed tosee the concentration dependent action of Aloe vera gel on platelet aggregation. Study Design:Comparative study. Setting: Post Graduate Medical Institute Lahore, Children Hospital Lahore.Duration of Study: One Year. Methodology: This study was conducted on healthy volunteersselected from staff and students of Ameer-ud-din Medical College. After determining baselineHb and platelet counts, PRP was prepared and then incubated with 2 different concentrationsof Aloe vera low (AVL) and Aloe vera high (AVH) for 30 minutes. Aggregation was stimulatedby adding the agonist arachidonic acid. Light transmission aggregometer was used to recordplatelet aggregation activity graphically for 3 minutes. Results: The data was analyzed usingSPSS version 20. Kruskal Wallis H test was performed to compare the platelet aggregation,which revealed that platelet aggregation with AVL and AVH were statistically significantly lower,amounting to 25.89% and 16.72% respectively as compared to 88.28% observed with control.Conclusion: This study has established in vitro anti-platelet effect of Aloe vera which is dosedependent.

In the current study, we analyzed the efficacy of manganese oxide nanoparticle (MnNP)-water dispersion as an immobilization matrix for bacterial tannase. The tannase-secreting Bacillus subtilis strain NJKL.tan.2 obtained from tannery effluent soil was subsequently purified and cloned in pET20b vector. The activity of MnNP-tan (tannase activated by manganese nanoparticles) was 1.51- and 3.5-fold higher at 20 °C and 80 °C, respectively, compared with the free enzyme. MnNP-tan decreased Km by 41.66 % and 3-fold, whereas free tannase showed two-fold and six-fold improvement in Kcat at 37 °C and 80 °C, respectively. MnNP-tan showed an increase in (half-life)t1/2and Ed by 13-fold and 50.05 units, respectively, at 80 °C, in contrast to the native enzyme. MnNP-tan retained its residual activity by 78.2 % at 37 °C and 34.24 % at 80 °C after 180 min of incubation when compared with untreated set. MnNP-tan retained 51 % of its activity after 120 days with the native enzyme losing ∼50 % functionality following 40 days of incubation. The MnNP-mediated tannase immobilization technique is being reported for the first time. The technique has numerous advantages due to the use of MnNP as a potential matrix for biomolecule immobilization, which can be further extended to immobilize other biocatalysts used in agro-industrial and lab-based applications.

// Pia Jensen 1 , Christa L. Myhre 2 , Pernille S. Lassen 1 , Athanasios Metaxas 2 , Asif M. Khan 2, 5 , Kate L. Lambertsen 2, 3, 4 , Alicia A. Babcock 2 , Bente Finsen 2, 4, * , Martin R. Larsen 1, * and Stefan J. Kempf 1 1 Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark 2 Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark 3 Department of Neurology, Odense University Hospital, Odense, Denmark 4 BRIDGE, Brain Research–Inter-Disciplinary Guided Excellence, Department of Clinical Research, University of Southern Denmark, Odense, Denmark 5 Current address: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark * These authors are equally contributed to these work Correspondence to: Stefan J. Kempf, email: stefank@bmb.sdu.dk Keywords: mTOR, neuroinflammation, alzheimer’s disease, post translational modification, LPS Received: March 21, 2017 Accepted: July 11, 2017 Published: July 21, 2017 ABSTRACT Neuroinflammation is a hallmark of Alzheimer’s disease and TNFα as the main inducer of neuroinflammation has neurodegenerative but also pro-regenerative properties, however, the dose-dependent molecular changes on signaling pathway level are not fully understood. We performed quantitative proteomics and phospho-proteomics to target this point. In HT22 cells, we found that TNFα reduced mitochondrial signaling and inhibited mTOR protein translation signaling but also led to induction of neuroprotective MAPK-CREB signaling. Stimulation of human neurons with TNFα revealed similar cellular mechanisms. Moreover, a number of synaptic plasticity-associated genes were altered in their expression profile including CREB . SiRNA-mediated knockdown of CREB in human neurons prior to TNFα stimulation led to a reduced number of protein/phospho-protein hits compared to siRNA-mediated knockdown of CREB or TNFα stimulation alone and countermeasured the reduced CREB signaling. In vivo data of TNFα knockout mice showed that learning ability did not depend on TNFα per se but that TNFα was essential for preserving the learning ability after episodes of lipopolysaccharide-induced neuroinflammation. This may be based on modulation of CREB/CREB signaling as revealed by the in vitro / in vivo data. Our data show that several molecular targets and signaling pathways induced by TNFα in neurons resemble those seen in Alzheimer’s disease pathology.

Insulin-like growth factor-1 (IGF-1) is a pleiotropic molecule with neurotrophic and immunomodulatory functions. Knowing the capacity of chronically activated microglia to produce IGF-1 may therefore show essential to promote beneficial microglial functions in Alzheimer's disease (AD). Here, we investigated the expression of IGF-1 mRNA and IGF-1 along with the expression of tumor necrosis factor (TNF) mRNA, and the amyloid-beta (A beta) plaque load in the hippocampus of 3- to 24-month-old APP(swe)/PS1(Delta E9) transgenic (Tg) and wild-type (WT) mice. As IGF-1, in particular, is implicated in neurogenesis we also monitored the proliferation of cells in the subgranular zone (sgz) of the dentate gyrus. We found that the A beta plaque load reached its maximum in aged 21- and 24 month-old APP(swe)/PS1(Delta E9) Tg mice, and that microglial reactivity and hippocampal IGF-1 and TNF mRNA levels were significantly elevated in aged APP(swe)/PS1(Delta E9) Tg mice. The sgz cell proliferation decreased with age, regardless of genotype and increased IGF-1/TNF mRNA levels. Interestingly, IGF-1 mRNA was expressed in subsets of sgz cells, likely neuroblasts, and neurons in both genotypes, regardless of age, as well as in glial-like cells. By double in situ hybridization these were shown to be IGF1 mRNA(+) CD11b mRNA(+) cells, i.e., IGF-1 mRNA-expressing microglia. Quantification showed a 2-fold increase in the number of microglia and IGF-1 mRNA-expressing microglia in the molecular layer of the dentate gyrus in aged APP(swe)/PS1(Delta E9) Tg mice. Double-immunofluorescence showed that IGF-1 was expressed in a subset of A beta plaque-associated CD11b(+) microglia and in several subsets of neurons. Exposure of primary murine microglia and BV2 cells to A beta(42) did not affect IGF-1 mRNA expression. IGF-1 mRNA levels remained constant in WT mice with aging, unlike TNF mRNA levels which increased with aging. In conclusion, our results suggest that the increased IGF-1 mRNA levels can be ascribed to a larger number of IGF-1 mRNA-expressing microglia in the aged APP(swe)/PS1(Delta E9) Tg mice. The finding that subsets of microglia retain the capacity to express IGF-1 mRNA and IGF-1 in the aged APP(swe)/PS1(Delta E9) Tg mice is encouraging, considering the beneficial therapeutic potential of modulating microglial production of IGF-1 in AD.

Introduction Treatment with selective serotonin reuptake inhibitors has been suggested to mitigate amyloid-β (Aβ) pathology in Alzheimer's disease, in addition to an antidepressant mechanism of action. Methods We investigated whether chronic treatment with paroxetine, a selective serotonin reuptake inhibitor, mitigates Aβ pathology in plaque-bearing double-transgenic amyloid precursor protein (APP)swe/presenilin 1 (PS1)ΔE9 mutants. In addition, we addressed whether serotonin depletion affects Aβ pathology. Treatments were assessed by measurement of serotonin transporter occupancy and high-performance liquid chromatography. The effect of paroxetine on Aβ pathology was evaluated by stereological plaque load estimation and Aβ42/Aβ40 ratio by enzyme-linked immunosorbent assay. Results Contrary to our hypothesis, paroxetine therapy did not mitigate Aβ pathology, and depletion of brain serotonin did not exacerbate Aβ pathology. However, chronic paroxetine therapy increased mortality in APPswe/PS1ΔE9 transgenic mice. Discussion Our results question the ability of selective serotonin reuptake inhibitor therapy to ameliorate established Aβ pathology. The severe adverse effect of paroxetine may discourage its use for disease-modifying purposes in Alzheimer's disease., Highlights • Chronic paroxetine treatment does not mitigate established amyloid-β pathology in APPswe/PS1ΔE9 mice. • Chronic paroxetine treatment increases the mortality of APPswe/PS1ΔE9 but not littermate wild-type mice. • Serotonergic depletion does not influence established amyloid-β pathology in APPswe/PS1ΔE9 mice.

For genetic transformation, an efficient in vitro regeneration system in frost sensitive potato genotypes was established in this research study. Two frost sensitive genotypes of potato viz, Kuroda and FD-51-5 with good agronomic features but sensitive to frost were used. Six callus formation media were investigated for callus formation, in which three levels of 2, 4-D (3.5 mg/L, 4.5 mg/L and 5.5 mg/L) alone and in combination with one level of NAA (0.25 mg/L) were used. Of these, four callogenesis media viz, CM1, CM2, CM3 and CM4 were proved to be best for callus formation while genotype Kuroda gave best response. So these four CM and genotype kuroda was selected for in vitro regeneration study in which five regeneration media (RM) were investigated. In these media three levels of BAP (2.75 mg/L, 3.75 mg/L, and 4.75 mg/L) and one level of NAA and kinetin (0.25 mg/L) were used in different combinations. For in vitro regeneration, two age groups of calli viz, 28 days old and 35 days old were studies to check the effect of calli age in vitro regeneration. These calli of genotype kuroda were induced on selected CM and then were put on these RM. Thereby results showed that twenty eight days old calli of genotype Kuroda from CM2 when shifted on RM4 gave maximum number of shoots.

No study in the past has examined the genetic diversity and zoonotic potential of Giardia duodenalis in dairy cattle in India. To assess the importance of these animals as a source of human G. duodenalis infections and determine the epidemiology of bovine giardiasis in India, fecal samples from 180 calves, heifers and adults and 51 dairy farm workers on two dairy farms in West Bengal, India were genotyped by PCR-RFLP analysis of the β-giardin gene of G. duodenalis followed by DNA sequencing of the nested PCR products. The overall prevalence of G. duodenalis in cattle was 12.2% (22/180), the infection being more prevalent in younger calves than in adult cattle. Zoonotic G. duodenalis Assemblage A1 was identified in both calves and workers although the most prevalent genotype detected in cattle was a novel Assemblage E subgenotype. These findings clearly suggest that there is a potential risk of zoonotic transmission of G. duodenalis infections between cattle and humans on dairy farms in India.

Few studies in the past have examined the genetic diversity and zoonotic potential of Cryptosporidium in dairy cattle in India. To assess the importance of these animals as a source of human Cryptosporidium infections, fecal samples from 180 calves, heifers and adults and 51 farm workers on two dairy farms in West Bengal, India were genotyped by PCR-RFLP analysis of the 18S rRNA gene of Cryptosporidium followed by DNA sequencing of the PCR products. Phylogenetic analysis was carried out on the DNA sequences obtained in the study and those available in GenBank. The overall prevalence of Cryptosporidium in cattle was 11.7% though the infection was more prevalent in younger calves than in adult cattle. The occurrence of Cryptosporidium parvum, Cryptosporidium bovis, Cryptosporidium ryanae and Cryptosporidium andersoni in cattle followed an age-related pattern. A Cryptosporidium suis-like genotype was also detected in a calf. Farm workers were infected with Cryptosporidium hominis, C. parvum and a novel C. bovis genotype. These findings clearly suggest that there is a potential risk of zoonotic transmission of Cryptosporidium infections between cattle and humans on dairy farms in India.

The susceptibility of the aging brain to neurodegenerative disease may in part be attributed to cellular aging of the microglial cells that survey it. We investigated the effect of cellular aging induced by telomere shortening on microglia by the use of mice lacking the telomerase RNA component (TERC) and design-based stereology. TERC knockout (KO) mice had a significantly reduced number of CD11b(+) microglia in the dentate gyrus. Because of an even greater reduction in dentate gyrus volume, microglial density was, however, increased. Microglia in TERC KO mice maintained a homogenous distribution and normal expression of CD45 and CD68 and the aging marker, ferritin, but were morphologically distinct from microglia in both adult and old wild-type mice. TERC KO mice also showed increased cellular apoptosis and impaired spatial learning. Our results suggest that individual microglia are relatively resistant to telomerase deficiency during steady state conditions, despite an overall reduction in microglial numbers. Furthermore, telomerase deficiency and aging may provide disparate cues leading to distinct changes in microglial morphology and phenotype.

Inhibition of heat shock protein 90 (HSP90) results in the degradation of oncoproteins that drive malignant progression and induce cell death, thus making HSP90 a potential target of cancer therapy. 6-Chloro-9-(4-methoxy-3, 5-dimethyl-pyridin-2-ylmethyl)-9H-purin-2-ylamine (BIIB021), a synthetic HSP90 inhibitor, exhibited promising antitumor activity in preclinical models. It is currently in phase II clinical trials for the oral treatment of breast cancer. The objective of this study was to obtain both quantitative and qualitative metabolic profiles of [(14)C]BIIB021 in rat, dog, and human liver microsomes and hepatocytes to provide support for in vivo safety and clinical studies. The metabolites of [(14)C]BIIB021 were identified using liquid chromatography-tandem mass spectrometry coupled with radiometric detection. BIIB021 was extensively metabolized in both liver microsomes and hepatocytes. The major oxidative metabolic pathways identified for all species were due to hydroxylation (M7) and O-demethylation (M2) of the methoxy-dimethylpyridine moiety. The majority of M7 in dog hepatocytes was further conjugated to form the glucuronide (M4). Oxidative dechlorination (M6), monooxygenation (M10), and oxidative N-dealkylation of the methoxy-dimethylpyridine moiety (M11 and M12) were observed as the minor metabolic pathways in hepatocytes of all three species. A glutathione conjugate (M18) was also identified in all species. Its formation was catalyzed, in part, by soluble glutathione transferase via direct displacement of the chlorine on the amino-chloropurine moiety. Subsequent minor secondary metabolites M13, M14, M15, and M17 were observed in human, dog, and rat hepatocytes. Results from incubations of BIIB021 with human recombinant cytochrome P450 (P450) isoforms and a P450 antibody inhibition study in human liver microsomes suggested that the formation of M7 is mainly catalyzed by CYP2C19 and CYP3A4, whereas the formation of minor metabolite M2 in human liver microsomes probably could be attributed to CYP3A4.

Optimization of stable in vitro regeneration system is indispensible to apply molecular approaches in crops. Due to its profound impact on genetic transformation studies, we established a reproducible and effectual in vitro regeneration system, in two whip smut (Ustilago scitaminea) susceptible genotypes viz., S-2003-us-127 and S-2003-us-371. Twelve callus formation media (CFM) were investigated for callus formation, in which four levels of 2,4-D (1 mg/L, 2 mg/L, 3 mg/L and 4 mg/L), two levels of BAP (0 mg/L and 0.5 mg/L) and two levels of kinetin (0 and 0.1 mg/l) were used in different combinations with basal MS Salt. CFM3 (3 mg/L 2,4-D), CFM4 (4 mg/L 2,4-D), CFM11 (3 mg/L 2,4-D and 0.1 mg/L kinetin) and CFM12 (4 mg/L 2,4-D and 0.1 mg/L kinetin) were proved to be the best for callus formation response in genotype S-2003-us-127. But in case of genotype S-2003-us-371, CFM3, CFM11 and CFM12 showed good response for callus induction. Among good responsive CFM, we selected CFM with low dose of 2,4-D (CFM3 and CFM11) for our regeneration experiment. For regeneration study, four regeneration media (RM) with different plant growth regulators viz., 2,4-D (0.1 mg/L), BAP (0.5 mg/L and 1 mg/L), kinetin (0.25 mg/L) and proline (250 mmg/L) plus MS salt were used. Calli of three (3) different ages, viz., 21 days, 28 days and 35 days from CFM3 and CFM11 were shifted on four regeneration media (RM). Among these four regeneration media, RM2 (0.1 mg/L 2,4-D and 1 mg/L BAP) gave an excellent regeneration response for genotype S-2003-us-127, when 28 days old calli from CFM11 were transferred to this. This combination was selected as combination of choice in genotype S-2003-us-127 for genetic transformation studies. Genotype S-2003-us-371, showed maximum regeneration, when 35 days old calli from CFM11 were kept on RM4 (0.1 mg/L 2,4-D, 1 mg/L BAP, 0.25 mg/L kinetin and 250 mg/L proline). Genetic stability of regenerated plants of selected media combination was confirmed with RAPD (PCR) analysis by using 5 RAPD primers.