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The present study was designed to estimate the active ingredients in the aqueous extract of fennel Foeniculum vulgare L. fruits and test the effects of different concentrations of the extract on serum estrogen and prolactin levels in female rats. The work was conducted to prepare the aqueous extract in the laboratory, while the secondary active substances in the extract were estimated using High-Performance Liquid Chromatography (HPLC) technology. The experiments were conducted in the animal house of the College of Science, Tikrit university,on a total of 12 adult albino virgin female rats divided into four groups, each having three rats.The aqueous extract of the fruit plant was administrated orally to animals at three concentrations (50%, 75%, and 100%) per day for 45 days. The fourth group was the control group that was left without treatment. Blood samples were obtained from the corner of the eye with a capillary tube and the serum was extracted to determine the hormone levels. The results showed that the aqueous extract of fennel fruits contains eleven active secondary metabolites, namely Eucalyptol, Terpinene, Anisole, Camphor, Anethole, Anisaldehyde, Apiole, a-pinene, Estragole, Fenchone, and Limonene, which seemed to played a role in regulating the levels of prolactin and estrogen hormones in rats treated with the extract. The results showed a direct relationship between the concentration of the fruit extract given to the animals and their serum levels of estrogen and prolactin. Increases in the levels of estrogen (44.56 ± 0.90 ng/ml) and prolactin (134.66 ± 0.80 ng/ml) were recorded using the concentration of 100% of the watery extract of Foeniculum vulgare L., as compared to the control group (126.33 ± 1.18; 6.37 ± 0.45 ng / ml, respectively). The results demonstrated the effective role of the aqueous extract of fennel fruits in stimulating estrogen and prolactin in female rats through their content of active compounds, which can be of great importance in stimulating the process of milk secretion in animals.

Background Helicobacter pylori (HP) infection has been associated with worse motor function in Parkinson's disease (PD). Objective We aimed to evaluate the effects of HP eradication on PD symptoms. Methods In this parallel-group, double-blind, randomized placebo-controlled, single-center trial, patients with PD with positive HP urea breath test and serology were block randomized (1:1) to receive standard eradication triple therapy or identically appearing placebo capsules for 1 week. Prespecified motor (International Parkinson and Movement Disorder Society Unified PD Rating Scale [MDS-UPDRS], timed tests, and home-based wearable sensor measurements), nonmotor (Leeds Dyspepsia Questionnaire and Montreal Cognitive Assessment), and quality-of-life (Parkinson's Disease Questionnaire-39) outcome measures were assessed at weeks 6, 12, 24, and 52. The primary outcome was the baseline-to-week 12 change in ON medication MDS-UPDRS motor scores. Lactulose-hydrogen breath testing for concomitant small intestinal bacterial overgrowth was performed at baseline and repeated at week 24, together with the urea breath test. Results A total of 310 patients were screened for eligibility and 80 were randomly assigned, of whom 67 were included in the full-analysis set (32 treatment group patients, 35 placebo patients). HP eradication did not improve MDS-UPDRS motor scores at week 12 (mean difference 2.6 points in favor of placebo, 95% confidence interval: -0.4 to 5.6, P = 0.089). There was no significant improvement in any motor, nonmotor, or quality-of-life outcome at weeks 12 and 52. Both the full-analysis and per-protocol analyses (based on eradication status) supported these conclusions. Small intestinal bacterial overgrowth status did not influence treatment results. Conclusions HP eradication does not improve clinical outcomes in PD, suggesting that there is no justification for routine HP screening or eradication with the goal of improving PD symptoms. © 2020 International Parkinson and Movement Disorder Society.

Pre‐release dietary treatment with methoprene, a juvenile hormone analogue, decreases the age at which male Queensland fruit flies mature and hence may decrease the post‐release delay until released sterile flies participate in sterile insect technique (SIT) programmes. However, if matings of young methoprene‐treated males are not effective at inducing sexual inhibition in their mates, then this treatment may not enhance SIT. The present study investigates efficacy of matings of methoprene‐treated males at inducing sexual inhibition in their mates. Methoprene incorporated into a diet of sugar and yeast hydrolysate (w/w 3:1) for 48 hr after emergence resulted in significantly increased male mating propensity when flies were

A study was conducted during the period of January to July, 2012 in the Entomology Field Laboratory, Bangladesh Agricultural University, Mymensingh to manage the Yellow Stem Borer (YSB) of rice eco-friendly following the Randomized Complete Block Design (RCBD) with four replications using the rice variety TN1. To keep in view this point, three insecticides viz. Dursban 20 EC, Convoy 25 EC, Belt 24 WG and three botanical extracts viz. Neem, Tobacco, Karanja extracts were used to compare their effectiveness against Yellow Stem Borer (YSB),Scirpophaga incertulus and also against natural enemies of Yellow Stem Borer (YSB) as Yellow Stem Borer (YSB),Scirpophaga incertulus causes dead heart and white head symptoms at vegetative and reproductive stage of rice respectively, the number of dead heart and white head symptoms were counted at different time interval viz. 7, 15, 21 days after spraying (DAS) to assess the effectiveness of the treatments. The chemicals and botanicals caused significant difference in their effects against Yellow Stem Borer (YSB). Among the chemicals Dursban 20 EC caused highest reduction in dead heart and white head symptoms and in case of botanicals Neem extracts caused highest reduction in the symptoms. The chemicals and botanicals were also evaluated for their side effects on natural enemies of rice. The botanicals were found less harmful than insecticides. Natural enemies like Lady Bird Beetle and Spider were abundant in the Neem extract sprayed rice field after several weeks of its application. The insecticides and botanicals reduced the infestation of Yellow Stem Borer (YSB), Scirpophaga incertulus and thereby significantly influenced the yield performance of rice. Dursban 20 EC treated plot showed highest yield (1.80 Kg/ plot) and Neem extract treated plot showed the yield 1.40 Kg/ plot. Considering the efficacy and eco-friendly nature of Neem extracts it could be considered as an effective botanicals in successful management of the pest Yellow Stem Borer (YSB), Scirpophaga incertulus of rice.

HIV-1 replicates in cells that express a wide array of innate immune sensors and may do so simultaneously with other pathogens. How a coexisting innate immune stimulus influences the outcome of HIV-1 sensing, however, remains poorly understood. Here, we demonstrate that the activation of a second signaling pathway enables a cyclic GMP-AMP synthase (cGAS)-dependent type I interferon (IFN-I) response to HIV-1 infection. We used RNA sequencing to determine that HIV-1 alone induced few or no signs of an IFN-I response in THP-1 cells. In contrast, when supplemented with suboptimal levels of bacterial lipopolysaccharide (LPS), HIV-1 infection triggered the production of elevated levels of IFN-I and significant upregulation of interferon-stimulated genes. LPS-mediated enhancement of IFN-I production upon HIV-1 infection, which was observed in primary macrophages, was lost by blocking reverse transcription and with a hyperstable capsid, pointing to viral DNA being an essential immunostimulatory molecule. LPS also synergistically enhanced IFN-I production by cyclic GMP-AMP (cGAMP), a second messenger of cGAS. These observations suggest that the DNA sensor cGAS is responsible for a type I IFN response to HIV-1 in concert with LPS receptor Toll-like receptor 4 (TLR4). Small amounts of a TLR2 agonist also cooperate with HIV-1 to induce type I IFN production. These results demonstrate how subtle immunomodulatory activity renders HIV-1 capable of eliciting an IFN-I response through positive cross talk between cGAS and TLR sensing pathways. IMPORTANCE Innate immune activation is a hallmark of HIV-1 pathogenesis. Thus, it is critical to understand how HIV-1 infection elicits innate immune responses. In this work, we show that HIV-1 infection of macrophages leads to a robust type I interferon (IFN) production only when a second signaling event is initiated by a coexisting immunostimulatory molecule. Our results show that HIV-1 infection alone is not sufficient for triggering a strong IFN response. We find that bacterial membrane components, which are recognized by endosomal innate sensors, enable production of elevated levels of IFNs and significant upregulation of interferon-stimulated genes upon HIV-1 infection. This IFN response is dependent on viral DNA synthesis and prevented by a stable capsid, pointing to an essential role for a DNA sensing molecule. These observations provide new insights into how different innate immune recognition pathways synergize during HIV-1 infection and determine the outcome of innate responses.

Queensland fruit flies Bactrocera tryoni (Froggatt) have a long adult maturation phase which, together with high mortality rates, can substantially reduce number of released flies that survive to mature and contribute to sterile insect technique (SIT) programmes. This constraint on SIT can potentially be addressed by incorporating methoprene, a juvenile hormone analogue, into an adult diet of sugar and yeast hydrolysate for 2 days after emergence. Methoprene treatments have been found to accelerate sexual development of male Queensland fruit fly, resulting in increased mating propensity of 5–7-day-old males in no-choice laboratory trials. Before considering deployment of methoprene as a pre-release treatment in SIT, it is necessary to demonstrate mating competitiveness and compatibility of methoprene-treated flies under field-like conditions. In the present study, we assessed whether methoprene treatment increases ability of sterile males (5 and 7 days old) to compete with mature (wild or laboratory) males for matings with mature (wild or laboratory) females in field cages. We also investigated mating compatibility to test for sexual isolation between sterile flies and mature (wild or laboratory) fertile flies. In mating competitiveness tests, methoprene-treated males of either age outperformed mature wild or laboratory males for matings with mature wild or laboratory females, respectively. Untreated 5- and 7-day-old males were less competitive than mature wild or laboratory mature males and hence had lower relative sterility indexes. Methoprene-treated males mated earlier in the evening and continued mating for longer than untreated sterile males and mature wild or laboratory males. In mating compatibility trials, methoprene-treated males mated equally with methoprene-treated females and mature females, whereas methoprene-treated females tended to mate more often with mature males than with methoprene-treated males. However, untreated flies of both sexes exhibited substantial sexual isolation. Pairings that comprised methoprene-treated males and mature females had shorter mating latency and longer copulations than other pairings. Unlike males, methoprene-treated females did not exhibit changes in mating latency or duration. Overall, the present study supports the use of pre-release dietary methoprene treatment in Queensland fruit fly SIT.

Sterile insect technique (SIT) is an environmentally benign pest management technique that involves releasing millions of sterile insects to suppress reproduction of pest populations. Many fruit flies, including Queensland fruit fly (Bactrocera tryoni Froggatt, ‘Q-fly’), have long adult maturation periods such that pre-maturation mortality can greatly reduce abundance of sexually active sterile males and impede SIT efficacy. Q-fly is the most difficult and costly challenge to market access for Australia’s horticulture industries, and has been targeted for intensive use of SIT program. We here demonstrate potential of pre-release caffeine supplements as a novel means to accelerate sexual maturation in male Q-fly. In mating trials, analytical caffeine was very effective at accelerating sexual maturation, while no positive effects of caffeine-containing instant coffee or guarana supplements were detected. In parallel, development of testes and ejaculatory apodemes was accelerated in males provided analytical caffeine but not instant coffee or guarana. High doses of guarana and instant coffee reduced longevity while even the highest doses of analytical caffeine did not affect longevity. Pre-release caffeine supplements promote sexual maturation in Q-flies, and similar benefits are expected in other fruit flies having long adult maturation periods.

Larval diets used for artificial rearing can have a significant effect on insect biology. The Queensland fruit fly (aka ‘Qfly’), Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), is one of the greatest challenges for fruit growers in Australia. The sterile insect technique (SIT) is being developed to manage outbreaks in regions that remain free of Qfly and to reduce populations in regions where this species is endemic. Factory scale rearing is essential for SIT; however, artificial larval diets are known to affect the microbiome of Qfly, which may then affect fly performance. In this study, high-throughput Illumina sequencing was used to assess the Qfly microbiome in colonies reared, for five generations from nature, on two common artificial diets (carrot and gel). At generation five (G5), the microbiome was assessed in larvae, pupae, adult males and adult females and standard fly quality control parameters were assessed together with additional performance measures of mating propensity and survival under nutritional stress. At the genus level, bacterial communities were significantly different between the colonies reared on the two larval diets. However, communities converged at Phyla to family taxonomic levels. Interestingly, pathogenic bacterial genera (e.g., Morganella, Citrobacter, Providencia, Burkholderia) were highly abundant in all developmental stages of Qfly reared on the gel diet, when compared to the carrot diet. Despite abundance of these commonly pathogenic genera, a greater percentage of egg hatching, heavier pupal weight and a higher percentage of fliers were found in the Qfly reared on the gel diet. Mating propensity and survival under nutritional stress was similar for adult Qfly that had been reared on the two larval diets. Overall, our findings demonstrate that the artificial larval diet strongly influences the microbiome and quality control measures of Qfly, with likely downstream effects on performance of flies released in SIT programs.

Double-stranded RNAs produced during viral infections serve as pathogen-associated molecular patterns (PAMPs) and bind pattern recognition receptors to stimulate IFN production. RNase L is an IFN-regulated endoribonuclease that is activated in virus-infected cells and cleaves single-stranded viral and cellular RNAs. The RNase L-cleaved dsRNAs signal to Rig-like helicases to amplify IFN production. This study identifies a novel role of antiviral stress granules induced by RNase L as an antiviral signaling hub to coordinate the RNA ligands with cognate receptors to mount an effective host response during viral infections., Virus infection leads to activation of the interferon (IFN)-induced endoribonuclease RNase L, which results in degradation of viral and cellular RNAs. Both cellular and viral RNA cleavage products of RNase L bind pattern recognition receptors (PRRs), like retinoic acid-inducible I (Rig-I) and melanoma differentiation-associated protein 5 (MDA5), to further amplify IFN production and antiviral response. Although much is known about the mechanics of ligand binding and PRR activation, how cells coordinate RNA sensing with signaling response and interferon production remains unclear. We show that RNA cleavage products of RNase L activity induce the formation of antiviral stress granules (avSGs) by regulating activation of double-stranded RNA (dsRNA)-dependent protein kinase R (PKR) and recruit the antiviral proteins Rig-I, PKR, OAS, and RNase L to avSGs. Biochemical analysis of purified avSGs showed interaction of a key stress granule protein, G3BP1, with only PKR and Rig-I and not with OAS or RNase L. AvSG assembly during RNase L activation is required for IRF3-mediated IFN production, but not IFN signaling or proinflammatory cytokine induction. Consequently, cells lacking avSG formation or RNase L signaling produced less IFN and showed higher susceptibility during Sendai virus infection, demonstrating the importance of avSGs in RNase L-mediated host defense. We propose a role during viral infection for RNase L-cleaved RNAs in inducing avSGs containing antiviral proteins to provide a platform for efficient interaction of RNA ligands with pattern recognition receptors to enhance IFN production to mount an effective antiviral response. IMPORTANCE Double-stranded RNAs produced during viral infections serve as pathogen-associated molecular patterns (PAMPs) and bind pattern recognition receptors to stimulate IFN production. RNase L is an IFN-regulated endoribonuclease that is activated in virus-infected cells and cleaves single-stranded viral and cellular RNAs. The RNase L-cleaved dsRNAs signal to Rig-like helicases to amplify IFN production. This study identifies a novel role of antiviral stress granules induced by RNase L as an antiviral signaling hub to coordinate the RNA ligands with cognate receptors to mount an effective host response during viral infections.

Juvenile hormone is an important regulator of sexual development in insects, and application of methoprene, a juvenile hormone analogue, together with access to a protein-rich diet, has been found to accelerate sexual maturation of several tephritid fruit fly species including Queensland fruit fly Bactrocera tryoni (‘Q-fly’). Such accelerated development is a potentially valuable means to increase participation of released males in sterile insect technique programs. However, there is a risk that benefits of accelerated maturation might be countered by increased vulnerability to starvation and desiccation. The present study investigates this possibility. After emergence, flies were treated with three levels of methoprene (0, 0.05%, and 0.5%) incorporated into a diet of sugar and yeast hydrolysate for two days after emergence. Survival of groups and individual flies was assessed under conditions of food stress, food and water stress, and ad libitum access to diet, and survival of individual flies was also assessed under desiccation stress. Most flies provided ad libitum access to diet were still alive at 7 days, whereas all stressed flies died within 4 days. Desiccation stressed flies had the shortest survival followed by food and water stress, and then food stress. Methoprene supplements increased susceptibility of flies to each stress. Flies subjected to food and water stress had the least lipid reserves at death, whereas flies subjected to desiccation stress retained the least water reserves. To investigate mechanisms that might underlie reduced survival under stress; we also quantified activity level of flies that were subjected to food and water stress and desiccation stress. Activity level was greater for flies provided methoprene, but did not vary with stress type or sex, suggesting that increased vulnerability of flies to stress is related to elevated metabolism associated with elevated activity. Deleterious effects of methoprene supplements on stress tolerance indicate a need for careful consideration of the conditions that will be encountered by flies in the field before deploying methoprene as a pre-release treatment in Q-fly sterile insect technique programs.

Sterile insect technique (SIT) is an environmentally benign pest management technique that relies on released sterile male insects mating with, and curtailing reproduction of, wild females. However, for species with high mortality rates and long adult maturation phases, a large proportion of the released insects can die before maturing and so fail to contribute to SIT. To counter this problem, inclusion of yeast hydrolysate in pre-release diets and treatment of pupae or adults with methoprene, a juvenile hormone analogue, have been investigated as means of accelerating development of some fruit flies, including Queensland fruit fly, Bactrocera tryoni (Froggatt) (‘Q-fly’). Methoprene has most often been administered topically in acetone solution, which is toxic, flammable, and impractical for operational settings. As a practical alternative, we incorporated methoprene (0, 0.05, 0.1, and 0.5%) into Q-fly adult diet of sugar only or sugar mixed with yeast hydrolysate for 2 days, and then provided sugar only for the rest of the trial period. Mating performance of males and females was tested from 4 to 30 days of age. Flies provided sugar mixed with yeast hydrolysate had increased mating propensity in comparison with flies that were provided sugar only. At all ages and for both diets, all methoprene doses increased male mating probability. Methoprene treatment did not affect copula latency of males that received yeast hydrolysate, but males that received only sugar mated earlier if they had received 0.05% methoprene. Methoprene treatment of males was also associated with longer copulations, which may affect fertility of females that later remate. Females differed from males in that methoprene treatment did not significantly affect mating probability or latency, but resembled males in that methoprene treatment resulted in longer copulations. Sex differences in response to methoprene may lead to male-biased operational sex ratio when bisex Q-fly strains are used in SIT. Yeast hydrolysate increased longevity of both males and females, but methoprene treatment did not affect longevity. Overall, findings of the present study indicate that Q-fly sexual maturation can be accelerated, and SIT might hence be enhanced, by incorporation of methoprene and yeast hydrolysate in pre-release diet.

An experiment was conducted in the Field Laboratory, Department of Entomology, (BAU), Mymensingh on the varietal preference of okra jassid, Amrasca devastans (Dist.) using 14 okra varieties viz. F 1 Tamanna, Hybrid Okra Gunyon, Okra F 1 Green Soft, OK 285, Okra F 1 Green Star 33, F 1 Hybrid Fresh Power, F 1 Hybrid Sarosh 3, Hybrid Okra (MDR), Okra Nabil, BARI-1 (Samrat), Hybrid Chamak and Local variety during March to June 2014. The results indicated that host preference of jassid differed significantly among the 14 varieties tested. Three varieties viz. Hybrid Chamak, Arka Anamika and Kolatia were the least preferred by jassid. Five varieties viz. Okra Nabil, Hybrid Okra (MDR), F 1 Hybrid Fresh Power, Local variety, and OK 285 were comparatively highly preferred and rest of the six varieties viz. Okra F 1 Green Star 33, BARI-1 (Samrat), F 1 Tamanna, F 1 Hybrid Sarosh 3, Hybrid Okra Gunyon, and Okra F 1 Green Soft were moderately preferred by jassid. This variation of preference of okra jassid might be due to different morphological and physiological characters of selected varieties. It could be concluded that comparatively resistant varieties selected from the present study might be incorporated to develop of IPM package for jassid. J. Bangladesh Agril. Univ. 15(2): 227-233, December 2017

Methoprene supplements added to diets of yeast hydrolysate and sugar promote early expression of sexual behaviour and mating in male Queensland fruit fly (Bactrocera tryoni; 'Q-fly') and show promise as a pre-release treatment for sterile insect technique programs. Currently it is not known whether the early mating behaviour of methoprene-treated male Q-flies is only behavioural or is coupled with accelerated development of reproductive organs. Accordingly, the present study investigates whether incorporation of methoprene into diets of yeast hydrolysate and sugar (1:3) or sugar alone, accelerate development of testes, ejaculatory apodeme, and accessory glands in male Q-flies and ovaries in females. All organs increased in size as the flies aged and matured, and development rate of all organs was far greater when the flies were provided yeast hydrolysate in addition to sugar. Incorporation of methoprene into diets containing yeast hydrolysate was found to strongly accelerate development of testes and ejaculatory apodeme, but not accessory glands, in males. In the absence of yeast hydrolysate, methoprene treatment had only a modest effect on male organ development. In contrast to males, development of ovaries in female Q-flies did not respond to dietary methoprene supplements, regardless of whether they were fed yeast hydrolysate and sugar or sugar alone. These findings of diet-dependent effects of methoprene supplements on reproductive organs are a close match to previous studies investigating effects of methoprene supplements on mating behaviour. Overall, methoprene supplements substantially enhance the positive effects of protein rich adult diet on the early expression of sexual behaviour and accelerate development of reproductive organs in male, but not female, Q-flies. Methoprene supplements added to pre-release diets of yeast hydrolysate and sugar show promise as a means of accelerating reproductive development of Q-flies released in sterile insect technique programs, and may also bias operational sex ratio in favour of males.

The HIV-1 capsid executes essential functions that are regulated by capsid stability and host factors. In contrast to increasing knowledge on functional roles of capsid-interacting host proteins during postentry steps, less is known about capsid stability and its impact on intracellular events. Here, using the antiviral compound PF-3450074 (PF74) as a probe for capsid function, we uncovered a novel phenotype of capsid stability that has a profound effect on innate sensing of viral DNA by the DNA sensor cGAS. A single mutation, R143A, in the capsid protein conferred resistance to high concentrations of PF74, without affecting capsid binding to PF74. A cell-free assay showed that the R143A mutant partially counteracted the capsid-destabilizing activity of PF74, pointing to capsid stabilization as a resistance mechanism for the R143A mutant. In monocytic THP-1 cells, the R143A virus, but not the wild-type virus, suppressed cGAS-dependent innate immune activation. These results suggest that capsid stabilization improves the shielding of viral DNA from innate sensing. We found that a naturally occurring transmitted founder (T/F) variant shares the same properties as the R143A mutant with respect to PF74 resistance and DNA sensing. Imaging assays revealed delayed uncoating kinetics of this T/F variant and the R143A mutant. All these phenotypes of this T/F variant were controlled by a genetic polymorphism located at the trimeric interface between capsid hexamers, thus linking these capsid-dependent properties. Overall, this work functionally connects capsid stability to innate sensing of viral DNA and reveals naturally occurring phenotypic variation in HIV-1 capsid stability. IMPORTANCE The HIV-1 capsid, which is made from individual viral capsid proteins (CA), is a target for a number of antiviral compounds, including the small-molecule inhibitor PF74. In the present study, we utilized PF74 to identify a transmitted/founder (T/F) strain that shows increased capsid stability. Interestingly, PF74-resistant variants prevented cGAS-dependent innate immune activation under a condition where the other T/F strains induced type I interferon. These observations thus reveal a new CA-specific phenotype that couples capsid stability to viral DNA recognition by cytosolic DNA sensors.

Queensland fruit flies Bactrocera tryoni (‘Q-fly’) have long adult prereproductive development periods, which can present challenges for sterile insect technique (SIT) programs. Holding the sterile flies in release facilities is expensive for control programs. Alternatively, releases of sexually immature males can lead to substantial mortality of sterile males before they mature. Recent studies have reported effectiveness of dietary supplementation with a mosquito larvicide (NOMOZ) that contains S-methoprene, a juvenile hormone analogue, for accelerating sexual development of fertile Q-fly males. However, it is not known whether effects on sterile flies are comparable to effects on fertile flies, or whether effects of methoprene-containing larvicide are comparable to effects of analytical standard methoprene such has been used in most studies. Here we address both knowledge gaps, investigating the effects of analytical standard methoprene and NOMOZ mixed with food and provided for 48 h following emergence on sexual development and longevity of fertile and sterile Q-flies. Compared with controls, fertile and sterile male Q-flies that were provided diets supplemented with methoprene from either source exhibited substantially accelerated sexual development by 2–3 d and longer mating duration. Unlike males, females did not respond to methoprene treatment. Although fertile and sterile flies were generally similar in sexual development and response to methoprene treatment, sterile flies of both sexes tended to have shorter copula duration than fertile flies. Neither methoprene supplements nor sterilization affected longevity of flies. The present study confirms effectiveness of dietary methoprene supplements in accelerating sexual development of both fertile and sterile male (but not female) Q-flies, and also confirms that low-cost mosquito larvicides that contain methoprene can achieve effects similar to those for high-cost analytical grade methoprene as prerelease supplements for Q-fly SIT.

This study aims to determine the effect of alkaline solution (NaOH) and Ethanol solution (C2H5OH) with different soaking times on the tensile strength of agave fiber and bamboo fiber composite on the tensile strength with diverse percentages of solution, and to determine the personal frequency and stiffness of this composite in diverse fiber directions, both numerically and experimentally. The research method was tensile strength test on the combination of agave fiber and bamboo fiber which was treated with alkaline solution (NaOH) and ethanol solution (C2H5OH) with percentages of solution of 10%, 20%, 30%, and 40%, for 2 hours and 4 hours immersion. Furthermore, the vibration test on the agave fiber and bamboo fiber composite was also done with a variation of four position placements of exciter. A simple pinch pedestal was used to get the value of personal frequency and stiffness of the composite beams made from agave fiber and bamboo fiber. The target to be achieved in the study is to produce composite beams with good mechanical properties.This study aims to determine the effect of alkaline solution (NaOH) and Ethanol solution (C2H5OH) with different soaking times on the tensile strength of agave fiber and bamboo fiber composite on the tensile strength with diverse percentages of solution, and to determine the personal frequency and stiffness of this composite in diverse fiber directions, both numerically and experimentally. The research method was tensile strength test on the combination of agave fiber and bamboo fiber which was treated with alkaline solution (NaOH) and ethanol solution (C2H5OH) with percentages of solution of 10%, 20%, 30%, and 40%, for 2 hours and 4 hours immersion. Furthermore, the vibration test on the agave fiber and bamboo fiber composite was also done with a variation of four position placements of exciter. A simple pinch pedestal was used to get the value of personal frequency and stiffness of the composite beams made from agave fiber and bamboo fiber. The target to be achieved in the study is to produce composite ...

Cleavage and polyadenylation specificity factor subunit 6 (CPSF6), a host factor that interacts with the HIV-1 capsid (CA) protein, is implicated in diverse functions during the early part of the HIV-1 life cycle, including uncoating, nuclear entry, and integration targeting. Preservation of CA binding to CPSF6 in vivo suggests that this interaction is fine-tuned for efficient HIV-1 replication in physiologically relevant settings. Nevertheless, this possibility has not been formally examined. To assess the requirement for optimal CPSF6-CA binding during infection of primary cells and in vivo , we utilized a novel CA mutation, A77V, that significantly reduced CA binding to CPSF6. The A77V mutation rendered HIV-1 largely independent from TNPO3, NUP358, and NUP153 for infection and altered the integration site preference of HIV-1 without any discernible effects during the late steps of the virus life cycle. Surprisingly, the A77V mutant virus maintained the ability to replicate in monocyte-derived macrophages, primary CD4 + T cells, and humanized mice at a level comparable to that for the wild-type (WT) virus. Nonetheless, revertant viruses that restored the WT CA sequence and hence CA binding to CPSF6 emerged in three out of four A77V-infected animals. These results suggest that the optimal interaction of CA with CPSF6, though not absolutely essential for HIV-1 replication in physiologically relevant settings, confers a significant fitness advantage to the virus and thus is strictly conserved among naturally circulating HIV-1 strains. IMPORTANCE CPSF6 interacts with the HIV-1 capsid (CA) protein and has been implicated in nuclear entry and integration targeting. Preservation of CPSF6-CA binding across various HIV-1 strains suggested that the optimal interaction between CA and CPSF6 is critical during HIV-1 replication in vivo . Here, we identified a novel HIV-1 capsid mutant that reduces binding to CPSF6, is largely independent from the known cofactors for nuclear entry, and alters integration site preference. Despite these changes, virus carrying this mutation replicated in humanized mice at levels indistinguishable from those of the wild-type virus. However, in the majority of the animals, the mutant virus reverted back to the wild-type sequence, hence restoring the wild-type level of CA-CPSF6 interactions. These results suggest that optimal binding of CA to CPSF6 is not absolutely essential for HIV-1 replication in vivo but provides a fitness advantage that leads to the widespread usage of CPSF6 by HIV-1 in vivo .

The viral capsid of HIV-1 interacts with a number of host factors to orchestrate uncoating and regulate downstream events, such as reverse transcription, nuclear entry, and integration site targeting. PF-3450074 (PF74), an HIV-1 capsid-targeting low-molecular-weight antiviral compound, directly binds to the capsid (CA) protein at a site also utilized by host cell proteins CPSF6 and NUP153. Here, we found that the dose-response curve of PF74 is triphasic, consisting of a plateau and two inhibitory phases of different slope values, consistent with a bimodal mechanism of drug action. High PF74 concentrations yielded a steep curve with the highest slope value among different classes of known antiretrovirals, suggesting a dose-dependent, cooperative mechanism of action. CA interactions with both CPSF6 and cyclophilin A (CypA) were essential for the unique dose-response curve. A shift of the steep curve at lower drug concentrations upon blocking the CA-CypA interaction suggests a protective role for CypA against high concentrations of PF74. These findings, highlighting the unique characteristics of PF74, provide a model in which its multimodal mechanism of action of both noncooperative and cooperative inhibition by PF74 is regulated by interactions of cellular proteins with incoming viral capsids. IMPORTANCE PF74, a novel capsid-targeting antiviral against HIV-1, shares its binding site in the viral capsid protein (CA) with the host factors CPSF6 and NUP153. This work reveals that the dose-response curve of PF74 consists of two distinct inhibitory phases that are differentially regulated by CA-interacting host proteins. PF74's potency depended on these CA-binding factors at low doses. In contrast, the antiviral activity of high PF74 concentrations was attenuated by cyclophilin A. These observations provide novel insights into both the mechanism of action of PF74 and the roles of host factors during the early steps of HIV-1 infection.

Tephritid fruit flies are amongst the most damaging insect pests of horticulture globally. Some of the key fruit fly species are managed using the sterile insect technique (SIT), whereby millions of sterile males are released to suppress reproduction of pest populations. Male annihilation technique (MAT), whereby sex specific lures are used to attract and kill males, is often used to reduce wild male numbers before SIT programs commence, providing released sterile males an increased numerical advantage. Overall program efficacy might be improved if MAT could be deployed simultaneously with SIT, continuously depleting fertile males from pest populations and replacing them with sterile males. However, such 'male replacement' requires a means of suppressing attraction of released sterile males to lures used in MAT. Previous studies have found that exposure of some fruit flies to lure compounds as mature adults can suppress subsequent response to those lures, raising the possibility of pre-release treatments. However, this approach requires holding flies until after maturation for treatment and then release. The present study takes a novel approach of exposing immature adult male Queensland fruit flies (Bactrocera tryoni, or 'Qfly') to raspberry ketone (RK) mixed in food, forcing these flies to ingest RK at ages far younger than they would naturally. After feeding on RK-supplemented food for two days after emergence, male Qflies exhibited a reduction in attraction to cuelure traps that lasted more than 20 days. This approach to RK exposure is compatible with current practises, in which Qflies are released as immature adults, and also yields advantages of accelerated reproductive development and increased mating propensity at young ages.

The interferon antiviral pathways and prostate cancer genetics converge on a regulated endoribonuclease, RNase L. Positional cloning and linkage studies mapped Hereditary Prostate Cancer 1 (HPC1) to RNASEL. To date, there is no correlation of viral infections with prostate cancer, suggesting that RNase L may play additional roles in tumor suppression. Here, we demonstrate a role of RNase L as a suppressor of androgen receptor (AR) signaling, cell migration and matrix metalloproteinase activity. Using RNase L mutants, we show that its nucleolytic activity is dispensable for both AR signaling and migration. The most prevalent HPC1-associated mutations in RNase L, R462Q and E265X, enhance AR signaling and cell migration. RNase L negatively regulates cell migration and attachment on various extracellular matrices. We demonstrate that RNase L knockdown cells promote increased cell surface expression of integrin β1 which activates Focal Adhesion Kinase-Sarcoma (FAK-Src) pathway and Ras-related C3 botulinum toxin substrate 1-guanosine triphosphatase (Rac1-GTPase) activity to increase cell migration. Activity of matrix metalloproteinase (MMP)-2 and -9 is significantly increased in cells where RNase L levels are ablated. We show that mutations in RNase L found in HPC patients may promote prostate cancer by increasing expression of AR-responsive genes and cell motility and identify novel roles of RNase L as a prostate cancer susceptibility gene.

Statins are 3-Hydroxy-3-methylgutaryl CoA (HMG CoA) reductase inhibitors and mainly used in cardiovascular diseases. However, they are also widely known for their anticancer activities mediated by antiproliferative, proapoptotic, anti-invasive, and radio sensitizing properties. They are tested alone in several clinical trials for their anticancer activity and the response in not satisfactory due to high dose requirements in humans. Tumor inhibitory concentration of statins is 10-100 μM for different cancer cell lines when tested in vitro. But statins can cause anorexia and death in some individuals if concentration is reached to 20-25 μM in serum. So, there is high risk factor. To avoid these unwanted harmful effects, we can give statins in combination with other chemotherapeutic drugs for synergism. It will reduce the concentration of statins in dosage, and can avoid unwanted toxic effects. When used along with the other chemo therapeutic agents like prenylation inhibitors, NSAIDS and standard chemotherapeutic agents, they have shown much better results at lower doses with little or no toxicity.

To study the effect of short term (2 wk) zinc supplementation on hemoglobin and iron status of children with acute diarrhea. This study was a prospective, open label, single arm interventional trial conducted from June 2008 through October 2009 in a teaching hospital of North India. Three to sixty months old children presenting with acute diarrhea participated in the study. Subjects were supplemented with recommended doses of oral zinc gluconate for 2 wk. Changes in levels of hemoglobin, serum iron, total iron binding capacity, and serum ferritin were the main outcome measures. Sixty-two patients completed the study successfully. The prevalence of anemia before and after 2 wk of zinc supplementation remained unchanged. However, a small decline (p > 0.05) was observed in mean hemoglobin (from 8.95 ± 1.4 to 8.73 ± 1.43 g/dL), serum iron (79.56 ± 45.81 to 78.61 ± 44.41 μg/dL) and ferritin (84.77 ± 45.35 to 83.55 ± 44.10 ng/mL) levels. Total iron binding capacity increased from 331.60 ± 109.72 to 341.30 ± 119.90 μg/dL post supplementation (p > 0.05). Even though statistically insignificant, the small change observed in the levels of hemoglobin, and indicators of iron status following short term zinc supplementation might assume significance in some settings in developing countries where children receive short courses of zinc repeatedly for frequent diarrheal episodes.

The aim of this study was to explore the phytochemical composition, heavy metals analysis and the antibacterial activity of six medicinal plants i.e., Terminalia chebula Retz (fruits), Aegle marmelos L., (fruits), Curcuma longa L., (rhizomes), Syzygium aromaticum L., (flower buds), Piper nigrum L., (seeds), Cinnamomum cassia L., (barks) and its two remedial recipes (recipe 1 and 2) used against diarrhea obtained from the local herbal practitioners (Hakeems). A preliminary phytochemical screening of the above-mentioned plants extract in methanol, chloroform, n-hexane and distilled water revealed the presence of various constituents such as alkaloids, flavonoids, tannins and saponins by using standard procedures. The quantitative phytochemical studies shows that alkaloids, flavonoid and saponins were in maximum amount in Terminalia chebula. The concentration of Cd, Ni, Pb, Fe, Cr, Cu and Zn were investigated by using an atomic absorption spectrometer. The obtained analysis shows that Cr, Fe and Pb were present in the highest concentration in medicinal plants and their recipes. The antibacterial activities of the crude extract found in the recipes of methanol, chloroform, n-hexane and distilled water were analyzed by using agar well disc diffusion assay and minimum inhibitory concentration (MIC) by broth dilution method against four bacterial strains, namely, E. coli, Salmonella, Shigella and Methicillin-resistant Staphylococcus aureus (MRSA), respectively. The maximum zones of inhibition in methanol, water, chloroform and n-hexane extracts were seen in recipe 2 against Shigella (22.16 &plusmn, 0.47 mm), recipe 2 against Shigella (20.33 &plusmn, 0.24 mm), recipe 1 against Shigella (20.30 &plusmn, 0.29 mm) and recipe 2 against E. coli (30.23 &plusmn, 0.12 mm), respectively. Furthermore, the recipe extracts are more active against the tested bacterial strains than the extracts from individual plants. Therefore, it is concluded that the use of herbal plants and their recipes are the major source of drugs in a traditional medicinal system to cure different diseases.

Introduction: Conflicting reports are available on the relationship of Iron Deficiency Anaemia (IDA) and iron therapy with oxidative stress. Aim: To study the levels of markers of oxidative stress and antioxidant status in children with IDA and to assess the effect of iron therapy on the same. Materials and Methods: This prospective, single centre, hospital based study was a sub-study of a randomized controlled trial conducted in the Department of Paediatrics, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh in collaboration with the Department of Biochemistry (of the same institution) between October 2009 to February 2011. The sub-study was conducted in two parts: in the first part, levels of a biomarker of oxidative stress {Malondialdehyde (MDA)} and anti-oxidant enzymes {Superoxide Dismutase (SOD), Catalase (CAT), Glutathione Peroxidase (GPx)} were assessed and compared between 67 children with IDA and 31 non-anaemic controls; in the second part, the effect of oral iron (6mg/kg/day) for eight weeks on these markers was studied in a subset of 35 children with IDA. The Bivariate correlations procedure was used to compute pair wise associations for a set of variables. T-tests (Independent samples t-test/Paired sample t-test) and Non-parametric tests (Mann–Whitney test/Wilcoxon signed-rank test) were applied as applicable for normally and non-normally distributed data, respectively. Results: Levels of anti-oxidant enzymes were significantly lower (p

The actin cytoskeleton and its network of associated proteins constitute a physical barrier that viruses must circumvent to gain entry into cells for productive infection. The mechanisms by which the physical signals of infection are sensed by the host to activate an innate immune response are not well understood. The antiviral endoribonuclease RNase L is ubiquitously expressed in a latent form and activated upon binding 2-5A, a unique oligoadenylate produced during viral infections. We provide evidence that RNase L in its inactive form interacts with the actin-binding protein Filamin A to modulate the actin cytoskeleton and inhibit virus entry. Cells lacking either RNase L or Filamin A displayed increased virus entry which was exacerbated in cells lacking both proteins. RNase L deletion mutants that reduced Filamin A interaction displayed a compromised ability to restrict virus entry, supporting the idea of an important role for the RNase L-Filamin A complex in barrier function. Remarkably, both the wild type and a catalytically inactive RNase L mutant were competent to reduce virus entry when transfected into RNase L-deficient cells, indicating that this novel function of RNase L is independent of its enzymatic activity. Virus infection and RNase L activation disrupt its association with Filamin A and release RNase L to mediate its canonical nuclease-dependent antiviral activities. The dual functions of RNase L as a constitutive component of the actin cytoskeleton and as an induced mediator of antiviral signaling and effector functions provide insights into its mechanisms of antiviral activity and opportunities for the development of novel antiviral agents., IMPORTANCE Cells constantly face and sample pathogens on their outer surface. The actin cytoskeleton and interacting proteins associate with the cell membrane and constitute a barrier to infection. Disruption of the actin cytoskeleton allows viruses to enter the cell and induces innate immune responses to clear infections. The molecular mechanisms that link virus-induced physical perturbations to host defense pathways remain unclear. Our studies identified a novel interaction between the antiviral endoribonuclease RNase L and the actin-binding protein Filamin A that enhances host defense by preventing viral entry into naive cells. This role for RNase L is independent of its enzymatic function. Virus infection alters actin dynamics, disrupts the RNase L-Filamin A complex, and releases RNase L to mediate antiviral signaling and effector functions via its established nucleolytic activities. These dual roles for RNase L provide an efficient strategy to protect cells from infection and rapidly respond upon pathogen exposure.

Sushovita Mukherjee, Mohammad Adnan Siddiqui, Shubham Dayal, Yasmine Zakaria Ayoub, Krishnamurthy Malathi Department of Biological Sciences, University of Toledo, Toledo, OH, USA Abstract: Chronic inflammation of the prostate contributes to the increased risk of prostate cancer. Microbial pathogens in the prostate cause inflammation that leads to prostatitis and proliferative inflammatory atrophy frequently associated with the development of prostate cancer. Bacterial lipopolysaccharides and DNA mediate immune responses by engaging Toll-like receptor (TLR) 4 and 9, respectively. Synthetic oligodeoxynucleotides containing CpG motifs (CpG-ODN) mimic bacterial DNA and signal through TLR9 to initiate innate immune responses. Here, we show that stimulation of DU145, PC3, or LnCap prostate cancer cells by the TLR9 agonists, CpG-ODN, induces mRNA expression of IL-6, IL-8, CXCL1, IP-10, CCL5, and TGFβ. In addition, activity of matrix metalloproteinase (MMP)-9 and -2 and cell migration increased on CpG-ODN treatment. Induction of cytokines and chemokines was mediated by NF-ΚB activation and translocation to the nucleus. Treatment with epigallocatechin-3-gallate (EGCG), the major constituent of green tea, prior to CpG-ODN stimulation, inhibits cytokine and chemokine gene induction, activity of MMP-9 and -2, and cell migration. EGCG treatment sequesters the p65 subunit of transcription factor NF-ΚB in the cytoplasm and inhibits transcriptional activity of the NF-ΚB-driven promoter in response to CpG-ODN. Our results suggest that the ability of the TLR9 agonists, CpG-ODN, to induce cytokines, chemokines, and MMP activity, as well as suppression by EGCG are independent of the androgen receptor and p53 status of the cells. EGCG may provide protective effects against inflammation in the prostate and benefit prostate cancer treatment. Keywords: CpG-ODN, EGCG, inflammation, NF-ΚB

Genetic diversity among four indigenous breeds of sheep namely Mengali, Balochi, Beverigh and Harnai of Balochistan was detected by random amplified polymorphic DNA (RAPD) technique. Nineteen RAPD primers were initially applied and seventeen were chosen for further analysis, based on band pattern quality, reproducibility and presence of bands. An average of 92 RAPD fragments were obtained by using 17 primers and out of the 36 fragments (39.13%) illustrated monomorphism while, 56 bands (60.87%) were polymorphic in all the four sheep breeds. The number of bands amplified in all the sheep breeds ranged from 2 to 10. The highest number of polymorphic loci 40 was observed in the Mengali breed, while the lowest 28 was in Balochi. Further, 33 and 31 polymorphic loci were seen in Beverigh and Harnai breeds, respectively. The overall gene diversity was highest in the Mengali (0.1474) while the lowest in Balochi breed (0.0998). Results of genetic similarities showed closer proximity between Balochi and Beverigh (0.992), Balochi and Harnai (0.992), and between Beverigh and Harnai (0.996). The resemblance was observed between Mengali and Balochi (0.918), between Mengali and Beverigh breeds (0.931) and between Mengali and Harnai breeds (0.925). The high level of genetic similarity between Balochi, Beverigh and Harnai sheep indicated the close relationship that might be due to common habitat. Further the present study highlighted the presence of diversity among and within breeds that can be used in the selection or crossbreeding programs of sheep. The present study suggests that RAPD-PCR can effectively be used to determine the genetic distances among the sheep breeds. Key words: Genetic distance, polymorphism, random amplified Polymorphic DNA.

An experiment was conducted in Field Laboratory, Department of Entomology at Bangladesh Agricultural University, Mymensingh, during 2013 to manage the mango hopper,Idioscopus clypealisL, using three chemical insecticides, Imidacloprid (0.3%), Endosulfan (0.5%), and Cypermethrin (0.4%), and natural Neem oil (3%) with three replications of each. All the treatments were significantly effective in managing mango hopper in comparison to the control. Imidacloprid showed the highest efficacy in percentage of reduction of hopper population (92.50 ± 9.02) at 72 hours after treatment in case of 2nd spray. It also showed the highest overall percentage of reduction (88.59 ± 8.64) of hopper population and less toxicity to natural enemies including green ant, spider, and lacewing of mango hopper. In case of biopesticide, azadirachtin based Neem oil was found effective against mango hopper as 48.35, 60.15, and 56.54% reduction after 24, 72, and 168 hours of spraying, respectively, which was comparable with Cypermethrin as there was no statistically significant difference after 168 hours of spray. Natural enemies were also higher after 1st and 2nd spray in case of Neem oil.

Autophagy is a tightly regulated mechanism that mediates sequestration, degradation, and recycling of cellular proteins, organelles, and pathogens. Several proteins associated with autophagy regulate host responses to viral infections. Ribonuclease L (RNase L) is activated during viral infections and cleaves cellular and viral single-stranded RNAs, including rRNAs in ribosomes. Here we demonstrate that direct activation of RNase L coordinates the activation of c-Jun N-terminal kinase (JNK) and double-stranded RNA-dependent protein kinase (PKR) to induce autophagy with hallmarks as accumulation of autophagic vacuoles, p62(SQSTM1) degradation and conversion of Microtubule-associated Protein Light Chain 3-I (LC3-I) to LC3-II. Accordingly, treatment of cells with pharmacological inhibitors of JNK or PKR and mouse embryonic fibroblasts (MEFs) lacking JNK1/2 or PKR showed reduced autophagy levels. Furthermore, RNase L-induced JNK activity promoted Bcl-2 phosphorylation, disrupted the Beclin1-Bcl-2 complex and stimulated autophagy. Viral infection with Encephalomyocarditis virus (EMCV) or Sendai virus led to higher levels of autophagy in wild-type (WT) MEFs compared with RNase L knock out (KO) MEFs. Inhibition of RNase L-induced autophagy using Bafilomycin A1 or 3-methyladenine suppressed viral growth in initial stages; in later stages autophagy promoted viral replication dampening the antiviral effect. Induction of autophagy by activated RNase L is independent of the paracrine effects of interferon (IFN). Our findings suggest a novel role of RNase L in inducing autophagy affecting the outcomes of viral pathogenesis.

Miraculin is a taste modifying plant protein, which displays a peculiar property of being able to modify sour taste into sweet taste. Thus, there has been an increasing interest in miraculin due to this amazing property. In the present study, miraculin gene was introduced into the Miyagawa Wase Satsuma mandarin (Citrus unshiu Marc.) callus by Agrobacterum-mediated transformation to produce a citrus transgenic plant and express the recombinant miraculin protein under the control of 35S promoter. Satsuma mandarin is one of the choicest citrus varieties grown widely and commercially in Korea, especially in Jeju Island, and Japan. Expression of this protein in the transgenic plant resulted in the accumulation of a significant amount of the miraculin protein in the leaves. To investigate whether the expressed protein was correctly modified, the dimerization and N-glycosylation of recombinant miraculin in the transgenic plants were analyzed. The recombinant protein also showed a sufficient biological activity. These results open up a new way of expression system in woody plants such as citrus and can provide a suitable alternative for producing recombinant miraculin.

Type II chromosomal toxin-antitoxin (TA) modules consist of a pair of genes that encode two components: a stable toxin and a labile antitoxin interfering with the lethal action of the toxin through protein complex formation. Bioinformatic analysis of Streptococcus mutans UA159 genome identified a pair of linked genes encoding a MazEF-like TA. Our results show that S. mutans mazEF genes form a bicistronic operon that is cotranscribed from a σ70-like promoter. Overproduction of S. mutans MazF toxin had a toxic effect on S. mutans which can be neutralized by coexpression of its cognate antitoxin, S. mutans MazE. Although mazF expression inhibited cell growth, no cell lysis of S. mutans cultures was observed under the conditions tested. The MazEF TA is also functional in E. coli , where S. mutans MazF did not kill the cells but rather caused reversible cell growth arrest. Recombinant S. mutans MazE and MazF proteins were purified and were shown to interact with each other in vivo , confirming the nature of this TA as a type II addiction system. Our data indicate that MazF is a toxic nuclease arresting cell growth through the mechanism of RNA cleavage and that MazE inhibits the RNase activity of MazF by forming a complex. Our results suggest that the MazEF TA module might represent a cell growth modulator facilitating the persistence of S. mutans under the harsh conditions of the oral cavity.

Contagious caprine pleuropneumonia (CCPP) caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp) is a disease of goats which causes high morbidity and mortality and is reported in many countries of the world. There are probably no reports on the molecular prevalence of Mccp, Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in Balochistan and any other part of Pakistan. Thirty goats (n = 30) with marked respiratory symptoms were selected and procured from forty goat flocks in Pishin district of Balochistan in 2008. The genomic deoxyribonucleic acid (DNA) from the lung samples (n = 30) of the slaughtered goats was purified and subjected to polymerase chain reaction (PCR) assays for the presence of Mycoplasma mycoides cluster members and Mp. The PCR-RFLP (restriction fragment length polymorphism) was also used to further confirm the Mccp. Of the thirty lung samples 17 (56.67%) were positive for the molecular prevalence of Mcc, Mccp and Mp. In total the molecular prevalence was observed as 17.65% for Mccp (n = 3), 70.59% for Mcc (n = 12) and 11.76% for Mp (n = 2). The RFLP profile has also validated the PCR results of Mccp by yielding two bands of 190 and 126 bp. The results of PCR-RFLP coupled with the presence of fibrinous pleuropneumonia and pleurisy during postmortem of goats (n = 3) strongly indicated the prevalence of CCPP in this part of world. Moreover the prevalence of Mcc and Mp is also alarming in the study area. We report for the very first time the molecular prevalence of Mcc, Mccp, and Mp in the lung tissues of goats in the Pishin district of Balochistan, Pakistan.

Autophagy is a tightly regulated mechanism that mediates sequestration, degradation and recycling of cellular proteins, organelles and pathogens. Several proteins associated with autophagy regulate host responses to viral infections. Ribonuclease L (RNase L) is activated during viral infections and cleaves cellular and viral single-stranded RNAs, including rRNAs in ribosomes. We demonstrate that direct activation of RNase L coordinates the activation of c-Jun N-terminal kinase (JNK) and double-stranded RNA-dependent protein kinase (PKR) to induce autophagy with hallmarks as accumulation of autophagic vacuoles, p62(SQSTM1) degradation and conversion of Microtubule-associated Protein Light Chain 3-I (LC3-I) to LC3-II. Accordingly, treatment of cells with pharmacological inhibitors of JNK or PKR and mouse embryonic fibroblasts (MEFs) lacking JNK1/2 or PKR showed reduced autophagy levels. Furthermore, RNase L-induced JNK activity promoted Bcl-2 phosphorylation, disrupted the Beclin1-Bcl-2 complex and stimulated autophagy. Viral infection with Encephalomyocarditis virus (EMCV) or Sendai virus led to higher levels of autophagy in wild-type (WT) MEFs compared to RNase L knock out (KO) MEFs. Inhibition of RNase L-induced autophagy using Bafilomycin A1 or 3-methyladenine suppressed viral growth in initial stages; in later stages autophagy promoted viral replication dampening the antiviral effect. Induction of autophagy by activated RNase L is independent of the paracrine effects of interferon (IFN). Our findings suggest a novel role of RNase L in inducing autophagy affecting the outcomes of viral pathogenesis.