Localization and Expression of Hsp27 and αB-Crystallin in Rat Primary Myocardial Cells during Heat Stress In Vitro

Neonatal rat primary myocardial cells were subjected to heat stress in vitro, as a model for investigating the distribution and expression of Hsp27 and αB-crystallin. After exposure to heat stress at 42°C for different durations, the activities of enzymes expressed during cell damage increased in the supernatant of the heat-stressed myocardial cells from 10 min, and the pathological lesions were characterized by karyopyknosis and acute degeneration. Thus, cell damage was induced at the onset of heat stress. Immunofluorescence analysis showed stronger positive signals for both Hsp27 and αB-crystallin from 10 min to 240 min of exposure compared to the control cells. According to the Western blotting results, during the 480 min of heat stress, no significant variation was found in Hsp27 and αB-crystallin expression; however, significant differences were found in the induction of their corresponding mRNAs. The expression of these small heat shock proteins (sHsps) was probably delayed or overtaxed due to the rapid consumption of sHsps in myocardial cells at the onset of heat stress. Our findings indicate that Hsp27 and αB-crystallin do play a role in the response of cardiac cells to heat stress, but the details of their function remain to be investigated.     

Characterization and engineering of the Lrp/AsnC family regulator SACE_5717 for erythromycin overproduction in Saccharopolyspora erythraea

Journal of Industrial Microbiology & Biotechnology - In this work, we found that the Lrp/AsnC family protein SACE_5717 negatively regulated erythromycin biosynthesis in S. erythraea. Disruption...     

The Utility of Sentinel Lymph Node Biopsy in Papillary Thyroid Carcinoma with Occult Lymph Nodes

Background

The sentinel lymph node (SLN) is defined as the first draining node from the primary lesion, and it has proven to be a good indicator of the metastatic status of regional lymph nodes in solid tumors. The aim of this study was to evaluate the clinical application of SLN biopsy (SLNB) in papillary thyroid carcinoma (PTC) with occult lymph nodes.

Methods

From April 2006 to October 2012, 212 consecutive PTC patients were treated with SLNB using carbon nanoparticle suspension (CNS). Then, the stained nodes defined as SLN were collected, and prophylactic central compartment neck dissection (CCND) followed by total thyroidectomy or subtotal thyroidectomy were performed. All the samples were sent for pathological examination.

Results

There were 78 (36.8%) SLN metastasis (SLNM)-positive cases and 134 (63.2%) SLNM-negative cases. The sensitivity, specificity, positive and negative predictive values, and false-positive and false-negative rates of SLNB were 78.8%, 100%, 100%, 84.3%, 0%, and 21.2%, respectively. The PTC patients with SLNM were more likely to be male (48.2% vs. 32.7%, p = 0.039) and exhibited multifocality (52.6% vs. 33.3%, p = 0.025) and extrathyroidal extension (56.7% vs. 33.5%, p = 0.015). A greater incidence of non-SLN metastases in the central compartment was found in patients with SLNM (41/78, 52.6%) than in those without SLNM (21/134, 15.7%; p < 0.05). However, the SLNM-negative PTC patients with non-SLN metastases were more likely to be male (37.9% vs. 9.5%, p < 0.05).

Conclusions

The application of SLNB using CNS is technically feasible, safe, and useful, especially for male patients with co-existing multifocality and extrathyroidal extension. However, the sensitivity of SLNB must be improved and its false-negative rate reduced before it can be a routine procedure and replace prophylactic CCND. More attention should be paid to PTC patients (especially males) without SLNM for signs of non-SLN metastases.     

Blockade of gap junction hemichannel suppresses disease progression in mouse models of amyotrophic lateral sclerosis and Alzheimer's disease

Background

Glutamate released by activated microglia induces excitotoxic neuronal death, which likely contributes to non-cell autonomous neuronal death in neurodegenerative diseases, including amyotrophic lateral sclerosis and Alzheimer''s disease. Although both blockade of glutamate receptors and inhibition of microglial activation are the therapeutic candidates for these neurodegenerative diseases, glutamate receptor blockers also perturbed physiological and essential glutamate signals, and inhibitors of microglial activation suppressed both neurotoxic/neuroprotective roles of microglia and hardly affected disease progression. We previously demonstrated that activated microglia release a large amount of glutamate specifically through gap junction hemichannel. Hence, blockade of gap junction hemichannel may be potentially beneficial in treatment of neurodegenerative diseases.

Methods and Findings

In this study, we generated a novel blood-brain barrier permeable gap junction hemichannel blocker based on glycyrrhetinic acid. We found that pharmacologic blockade of gap junction hemichannel inhibited excessive glutamate release from activated microglia in vitro and in vivo without producing notable toxicity. Blocking gap junction hemichannel significantly suppressed neuronal loss of the spinal cord and extended survival in transgenic mice carrying human superoxide dismutase 1 with G93A or G37R mutation as an amyotrophic lateral sclerosis mouse model. Moreover, blockade of gap junction hemichannel also significantly improved memory impairments without altering amyloid β deposition in double transgenic mice expressing human amyloid precursor protein with K595N and M596L mutations and presenilin 1 with A264E mutation as an Alzheimer''s disease mouse model.

Conclusions

Our results suggest that gap junction hemichannel blockers may represent a new therapeutic strategy to target neurotoxic microglia specifically and prevent microglia-mediated neuronal death in various neurodegenerative diseases.     

Aspirin-induced heat stress resistance in chicken myocardial cells can be suppressed by BAPTA-AM in vitro

Our recent studies have displayed the protective functions of aspirin against heat stress (HS) in chicken myocardial cells, and it may be associated with heat shock proteins (HSPs). In this study, we further investigated the potential role of HSPs in the aspirin-induced heat stress resistance. Four of the most important HSPs including HspB1 (Hsp27), Hsp60, Hsp70, and Hsp90 were induced by aspirin pretreatment and were suppressed by BAPTA-AM. When HSPs were induced by aspirin, much slighter HS injury was detected. But more serious damages were observed when HSPs were suppressed by BAPTA-AM than those cells exposed to HS without BAPTA-AM, even the myocardial cells have been treated with aspirin in prior. Comparing to other HSPs, HspB1 presented the largest increase after aspirin treatments, 86-fold higher than the baseline (the level before HS). These findings suggested that multiple HSPs participated in aspirin’s anti-heat stress function but HspB1 may contribute the most. Interestingly, during the experiments, we also found that apoptosis rate as well as the oxidative stress indicators (T-SOD and MDA) was not consistently responding to heat stress injury as expected. By selecting from a series of candidates, myocardial cell damage-related enzymes (CK-MB and LDH), cytopathological tests, and necrosis rate (measured by flow cytometry assays) are believed to be reliable indicators to evaluate heat stress injury in chicken’s myocardial cells and they will be used in our further investigations.     

Co-enzyme Q10 and acetyl salicylic acid enhance Hsp70 expression in primary chicken myocardial cells to protect the cells during heat stress

Pulmonary arterial hypertension (PAH) is characterized by vasoconstriction and proliferative obstruction of pulmonary vessels, which promotes a progressive increase in pulmonary vascular resistance (PVR). The effect of exercise training on oxidative stress, metabolism, and markers of nitric oxide (NO) and endothelin-1 (ET-1) was analyzed in the lung tissue of rats with PAH induced by monocrotaline (MCT).Twenty-four Wistar rats were divided into four groups (5–7 animals): sedentary control (SC), sedentary MCT (SM), trained control (TC), and trained MCT (TM). The TC and TM groups participated in a treadmill training protocol (60% VO₂ max) for 5 weeks, 3 weeks of which were performed after the injection of MCT (60 mg/kg i.p.) or saline. MCT administration promoted an increase in PVR and right ventricle hypertrophy, and reduction of right ventricle systolic function assessed by echocardiography. These changes were not improved by exercise training. The activity of NO synthase was reduced in the animals of the TC, TM, and SM groups. No significant differences were found in total nitrite concentration and expression of endothelial NO synthase. Moreover, the TM group showed strong staining for iNOS and nitrotyrosine, suggesting an increase in oxidative stress in these animals. In parallel, reduced expression of type B ET-1 receptors was noticed in the SM and TM groups in comparison to controls. In conclusion, the aerobic training protocol was unable to mitigate changes in the metabolism of NO and ET-1, probably because of the disease severity in these animals, especially in the TM group.     

An Entomopathogenic Strain of Beauveria bassiana against Frankliniella occidentalis with no Detrimental Effect on the Predatory Mite Neoseiulus barkeri: Evidence from Laboratory Bioassay and Scanning Electron Microscopic Observation

Among 28 isolates of Beauveria bassiana tested for virulence against F. occidentalis in laboratory bioassays, we found strain SZ-26 as the most potent, causing 96% mortality in adults at 1×10⁷ mL⁻¹conidia after 4 days. The effect of the strain SZ-26 on survival, longevity and fecundity of the predatory mite Neoseiulus (Amblyseius) barkeri Hughes were studied under laboratory conditions. The bioassay results showed that the corrected mortalities were less than 4 and 8% at 10 days following inoculation of the adult and the larvae of the predator, respectively, with 1×10⁷ conidia mL⁻¹ of SZ-26. Furthermore, no fungal hyphae were found in dead predators. The oviposition and postoviposition durations, longevity, and fecundity displayed no significant differences after inoculation with SZ-26 using first-instar larvae of F. occidentalis as prey in comparison with untreated predator. In contrast, the preoviposition durations were significantly longer. Observations with a scanning electron microscope, revealed that many conidia were attached to the cuticles of F. occidentalis at 2 h after treatment with germ tubes oriented toward cuticle at 24 h, penetration of the insect cuticle at 36 h, and finally, fungal colonization of the whole insect body at 60 h. In contrast, we never observed penetration of the predator''s cuticle and conidia were shed gradually from the body, further demonstrating that B. bassiana strain SZ-26 show high toxicity against F. occidentalis but no pathogenicity to predatory mite.     

印度犀牛角的化学成分分析

本文以印度犀牛角为主要研究对象,利用傅里叶变换红外光谱仪、X射线荧光光谱仪、全自动氨基酸分析仪分析其红外光谱、化学成分等特征,旨在为犀牛角替代品、人工合成品的研究提供部分基础数据。结果表明：红外光谱中氨基酸、磷脂酸、牛磺酸等相关的吸收峰明显;犀牛角主要的无机成分为CaO、Fe₂O₃、K₂O、ZnO、SiO₂等;氨基酸含量为481.01 mg/g,药用氨基酸含量丰富,氨基酸组成与标准蛋白的贴近度为0.74。结果表明犀牛角具有较高的营养价值。     

Vitamin C mitigates heat damage by reducing oxidative stress,inducing HSP expression in TM4 Sertoli cells

Heat stress is a major stressor that can lead to male reproductive dysfunction. Sertoli cells play a crucial role in spermatogenesis by providing germ cells with structural and nutritional support, and contributing to blood–testis barrier formation. Vitamin C (Vc) is an antioxidant capable of neutralizing reactive oxygen species and preventing lipid peroxidation widely used because it is inexpensive and highly accessible. In the present study, we investigated the protective effect of Vc on TM4 cells following heat stress. Pretreatment with Vc could effectively inhibit apoptosis (p < 0.01), lipid peroxidation, and lactate dehydrogenase (LDH) activity. However, a significant increase in the malondialdehyde (MDA) level and LDH activity (p < 0.01) was observed in TM4 cells without Vc‐pretreatment, in conjunction with vacuole degeneration and karyopyknosis. In addition, both the messenger RNA and protein levels of CryAB, Hsp27, Hsp70, and Hsp110 substantially increased in the 3 and 12 hr recovery groups (p < 0.01). Vc also prevented microtubule aggregation following heat stress. These results suggest that pretreatment with Vc‐protected TM4 cells against heat stress by reducing the level of oxidative stress and inducing heat shock protein expression.     

Potential use of the fungus Beauveria bassiana against the western flower thrips Frankliniella occidentalis without reducing the effectiveness of its natural predator Orius sauteri (Hemiptera: Anthocoridae)

Orius sauteri (Poppius; Hemiptera: Anthocoridae) is an important predator of western flower thrips, Frankliniella occidentalis (Pergande; Thysanoptera: Thripidae). O. sauteri would be directly exposed to the entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin in the field should the fungus be used as a biopesticide. If the fungus were to negatively affect O. sauteri in agro-ecosystems, predation of F. occidentalis by O. sauteri may be limited. The present study was undertaken to evaluate the insecticidal activity of strain B.bassiana-RSB of B. bassiana, which is highly virulent to F. occidentalis, on the predator under laboratory conditions. Results showed that, regardless of the concentration applied to first instars, Bb-RSB was not insecticidal against O. sauteri, nor did direct applications affect the developmental rate of the predator. Significant differences in developmental rates and adult longevity were observed between O. sauteri that fed on Bb-RSB-infected F. occidentalis cadavers and those that fed on untreated thrips. Developmental time (from first instar to adult) increased from 0.3 to 0.7 days and adult longevity decreased by 0.8 to 1.2 days for predators fed thrips treated with low and high concentrations of strain Bb-RSB, respectively, compared with predators fed on untreated thrips. However, these differences were only 3–13% of mean values for the controls, suggesting that the effects of Bb-RSB on O. sauteri are relatively minor. These findings highlight the potential use of O. sauteri in combination with B. bassiana for the biocontrol of F. occidentalis, but field tests must be performed to confirm their compatible use.