Decreased dynamin-related protein 1-related mitophagy induces myocardial apoptosis in the aging heart.

An increase in cardiomyocyte apoptosis is the main contributor to the observed high morbidity of cardiac disease during aging. Mitochondria play important roles in cardiac apoptosis, and dynamin-related protein 1 (Drp1) is the critical factor that participates in mitochondrial fission and induces mitophagy to maintain mitochondria quality. However, whether Drp1 is involved in the increase of apoptosis in aging heart remains unclear. The purpose of this study was to determine whether Drp1 participates in inducing the apoptosis through regulating mitophagy in aging myocardium. To explore the effect of mitophagy and apoptosis in aging heart, we detected the expression of COX IV and the co-localization of COX IV and LC3 II, which reflect mitophagy, and measured adenosine triphosphate and reactive oxygen species contents, which reflect mitochondrial injury. Cell apoptosis was detected by measuring the activity of caspase-3 and the expression of cleaved caspase-3 and further confirmed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. The results showed an increase in apoptosis and a decrease in mitophagy in aging cardiomyocytes, and apoptosis was ameliorated after the induction of mitophagy by carbonyl cyanide m-chlorophenyl hydrazone (a mitophagy activator) in D-galactose (D-gal)-induced senescence H9c2 cells. To clarify the role of Drp1 in apoptosis, we knocked down Drp1 by transfecting si-Drp1, or overexpressed Drp1 in senescent cells, and then detected mitophagy, mitochondrial injury, and apoptosis. The data showed that downregulated Drp1 induces mitochondrial damage and apoptosis. In addition, to explore the regulatory relationship between Drp1 and phosphatase and tensin homologue (PTEN)-induced putative kinase 1 (PINK1)/Parkin-mediated mitophagy, we detected the expressions of PINK1 and Parkin after the overexpression of Drp1 in the D-gal group cells and found that Drp1-mediated mitophagy inhibited the PINK1/Parkin pathway in senescent cells. Our results demonstrated that insufficient Drp1 induces cardiomyocyte apoptosis by inhibiting mitophagy, and Drp1 affects the PINK1/Parkin pathway of mitophagy in the aging heart.

Expansion of Luminal Progenitor Cells in the Aging Mouse and Human Prostate.

Aging is associated with loss of tissue mass and a decline in adult stem cell function in many tissues. In contrast, aging in the prostate is associated with growth-related diseases including benign prostatic hyperplasia (BPH). Surprisingly, the effects of aging on prostate epithelial cells have not been established. Here we find that organoid-forming progenitor activity of mouse prostate basal and luminal cells is maintained with age. This is caused by an age-related expansion of progenitor-like luminal cells that share features with human prostate luminal progenitor cells. The increase in luminal progenitor cells may contribute to greater risk for growth-related disease in the aging prostate. Importantly, we demonstrate expansion of human luminal progenitor cells in BPH. In summary, we define a Trop2+ luminal progenitor subset and identify an age-related shift in the luminal compartment of the mouse and human prostate epithelium.

Genome-wide, integrative analysis implicates microRNA dysregulation in autism spectrum disorder.

Genetic variants conferring risk for autism spectrum disorder (ASD) have been identified, but the role of post-transcriptional mechanisms in ASD is not well understood. We performed genome-wide microRNA (miRNA) expression profiling in post-mortem brains from individuals with ASD and controls and identified miRNAs and co-regulated modules that were perturbed in ASD. Putative targets of these ASD-affected miRNAs were enriched for genes that have been implicated in ASD risk. We confirmed regulatory relationships between several miRNAs and their putative target mRNAs in primary human neural progenitors. These include hsa-miR-21-3p, a miRNA of unknown CNS function that is upregulated in ASD and that targets neuronal genes downregulated in ASD, and hsa_can_1002-m, a previously unknown, primate-specific miRNA that is downregulated in ASD and that regulates the epidermal growth factor receptor and fibroblast growth factor receptor signaling pathways involved in neural development and immune function. Our findings support a role for miRNA dysregulation in ASD pathophysiology and provide a rich data set and framework for future analyses of miRNAs in neuropsychiatric diseases.

Antibody detection tests for early diagnosis in tuberculous meningitis.

BACKGROUND: Tuberculous meningitis (TBM) is the most severe form of tuberculosis. Microbiological confirmation is rare and treatment is often delayed. Early diagnosis and immediate initiation of treatment are essential for effective TBM control. A systematic review was performed in this study to assess the diagnostic accuracy of detecting antibodies against Mycobacterium tuberculosis in the cerebrospinal fluid (CSF), according to standard methods. Test performance was summarized using a bivariate random-effects meta-analysis. METHODS: Studies were identified by a search of the literature, up to July 25, 2015, in the EMBASE and MEDLINE databases via Ovid SP and PubMed. The Cochrane Library was also searched for original, peer-reviewed molecular epidemiology studies that reported the diagnosis of TBM based on antibody detection in the CSF. RESULTS: Thirty-six articles (58 studies) were identified. The sensitivity of antibody detection was 0.75 (95% confidence interval (CI) 0.66-0.82), specificity was 0.98 (95% CI 0.96-0.99), and the area under the receiver operating characteristic curve (AUROC) was 0.97 (95% CI 0.95-0.98). By subgroup analysis, the detection of anti-M37Ra was the highest (AUROC 0.99, 95% CI 0.98-1.00), followed by anti-antigen 5 (AUROC 0.99, 95% CI 0.97-0.99) and anti-M37Rv (AUROC 0.97, 95% CI 0.95-0.98). CONCLUSIONS: For the early diagnosis of TBM based on antibodies in the CSF, the detection of anti-M37Ra, anti-antigen 5, or anti-M37Rv provides the greatest sensitivity and specificity.

The balance between capture and dissociation of presynaptic proteins controls the spatial distribution of synapses.

The location, size, and number of synapses critically influence the specificity and strength of neural connections. In axons, synaptic vesicle (SV) and active zone (AZ) proteins are transported by molecular motors and accumulate at discrete presynaptic loci. Little is known about the mechanisms coordinating presynaptic protein transport and deposition to achieve proper distribution of synaptic material. Here we show that SV and AZ proteins exhibit extensive cotransport and undergo frequent pauses. At the axonal and synaptic pause sites, the balance between the capture and dissociation of mobile transport packets determines the extent of presynaptic assembly. The small G protein ARL-8 inhibits assembly by promoting dissociation, while a JNK kinase pathway and AZ assembly proteins inhibit dissociation. Furthermore, ARL-8 directly binds to the UNC-104/KIF1A motor to limit the capture efficiency. Together, molecular regulation of the dichotomy between axonal trafficking and local assembly controls vital aspects of synapse formation and maintenance.

Chaperone-dependent mechanisms for acid resistance in enteric bacteria.

The extremely acidic environment of the mammalian stomach not only serves to facilitate food digestion but also acts as a natural barrier against infections of food-borne pathogens. Many pathogenic bacteria, such as enterohemorrhagic Escherichia coli, can breach this host defense and cause severe diseases. These pathogens have evolved multiple intricate strategies to overcome the bactericidal activity of acids. In particular, recent studies have uncovered the central roles of two periplasmic chaperones, HdeA and HdeB, in protecting enteric bacteria from extremely acidic conditions. Here, we review recent advances in the understanding of the acid resistance mechanisms of Gram-negative bacteria and focus on the mechanisms of HdeA and HdeB in preventing acid-induced protein aggregation and facilitating protein refolding following pH neutralization.

An Arf-like small G protein, ARL-8, promotes the axonal transport of presynaptic cargoes by suppressing vesicle aggregation.

Presynaptic assembly requires the packaging of requisite proteins into vesicular cargoes in the cell soma, their long-distance microtubule-dependent transport down the axon, and, finally, their reconstitution into functional complexes at prespecified sites. Despite the identification of several molecules that contribute to these events, the regulatory mechanisms defining such discrete states remain elusive. We report the characterization of an Arf-like small G protein, ARL-8, required during this process. arl-8 mutants prematurely accumulate presynaptic cargoes within the proximal axon of several neuronal classes, with a corresponding failure to assemble presynapses distally. This proximal accumulation requires the activity of several molecules known to catalyze presynaptic assembly. Dynamic imaging studies reveal that arl-8 mutant vesicles exhibit an increased tendency to form immotile aggregates during transport. Together, these results suggest that arl-8 promotes a trafficking identity for presynaptic cargoes, facilitating their efficient transport by repressing premature self-association.

Changes in lymphocyte subsets in the peripheral blood of patients with active pulmonary tuberculosis.

The aim of this study was to determine the percentage of lymphocyte subsets in peripheral blood in patients with active tuberculosis. A total of 21 patients with active tuberculosis and 15 healthy volunteers were included in the study. T-lymphocyte subsets, B-lymphocytes (CD19(+) cells), natural killer (NK) cells and cells positive for costimulatory molecules CD28 and CD152 were evaluated using flow cytometry. Patients with tuberculosis had a significantly decreased percentage of CD3(+) and CD3(+)CD4(+) cells, and a significantly decreased ratio of CD3(+)CD4(+) to CD3(+)CD8(+) cells compared with healthy controls. In contrast, the percentage of B-cells (CD19(+) cells), CD3(+)CD8(+) cells, CD28(+) cells, CD152(+) cells, and subpopulations of CD4(+)CD152(+), CD8(+)CD152(+) and CD8(+)CD28(+) T-cells were all significantly increased compared with healthy controls. There were no statistically significant differences in the percentages of NK cells or CD4(+)CD28(+) cells between patients and controls. These results indicate that patients with active tuberculosis have altered lymphocyte homeostasis.

Autophagic and apoptotic cell death in amniotic epithelial cells.

The aim of this paper is to determine if autophagic cell death is associated with apoptosis and whether it participates in the process of term amniotic rupture. Forty pieces of fresh term amnions, including twenty from a position near the margin of the placentas and twenty from the margin of the naturally ruptured part of the placentas in term gestation were collected, respectively. The amnions were examined by scanning electron microscopy (SEM) and amniotic epithelial (AE) cells were examined by transmission electron microscopy (TEM). Autophagic and apoptotic cell death (PCD) were assayed by laser scanning confocal microscopy (LSCM) or flow cytometry using monodansylcadaverin (MDC) and propidium iodide (PI) stain. BCL(2) and BAX were examined by immunoblotting. Under SEM the amniotic epithelia appeared normal in the position near the placenta. They had an atrophied appearance in the margin of their natural broken parts. In the AE cells PCD was divided into three subtypes by TEM: autophagic cell death with positive stains of MDC and PI; apoptotic cell death; and the mixed type. Quantitative detection showed that there were more death cells, including autophagic and apoptotic, in the AE cells near the ruptured parts than near the placentas. An increased expression of BAX and a decreased expression of BCL(2) protein in the AE cells near the broken margin were observed. Apoptotic and autophagic cell death by the intrinsic pathway are the basic event in the AE cell and they are involved in the cause of membrane rupture of the human amnion in term gestation.

Conserved amphiphilic feature is essential for periplasmic chaperone HdeA to support acid resistance in enteric bacteria.

The extremely acidic environment of the mammalian stomach (pH 1-3) represents a stressful challenge for enteric pathogenic bacteria, including Escherichia coli, Shigella and Brucella. The hdeA (hns-dependent expression A) gene was found to be crucial for the survival of these enteric bacteria under extremely low pH conditions. We recently demonstrated that HdeA is able to exhibit chaperone-like activity exclusively within the stomach pH range by transforming from a well-folded conformation at higher pH values (above pH 3) into an unfolded conformation at extremely low pH values (below pH 3). This study was performed to characterize the action mechanisms and underlying specific structural features for HdeA to function in this unfolded conformation. In the present study, we demonstrate that the conserved 'amphiphilic' feature of HdeA, i.e. the exposure of the conserved hydrophobic region and highly charged terminal regions, is essential for exhibiting chaperone-like activity under extremely low pH conditions. Mutations that disrupt this amphiphilic feature markedly reduced the chaperone-like activity of HdeA. The results also strongly suggest that this acid-induced chaperone-like activity of HdeA is crucial for acid resistance of the enteric bacteria. Moreover, our new understanding of this amphiphilic structural feature of HdeA helps to better interpret how this unfolded (disordered) conformation could be functionally active.

Scavenging of reactive oxygen species by a urinary preparation.

In the present paper, the antioxidant properties of a preparation of human urine (PHU) were evaluated by studying the ability of this drug to react with relevant biological oxidants such as superoxide anion radical (O2*-) and hydroxyl radical (OH*). In addition, its effect on lipid peroxidation was investigated in vitro and ex vivo. PHU is not a good scavenger of O2*-. However, it reacts rapidly with OH radicals with a second-order rate constant of 2.8 x 10(9)/M/sec. The studies on rat brain homogenates showed that PHU had an inhibitory effect, which was dependent on its concentration and the magnitude of lipid peroxidation. Ex vivo studies also showed that oral administration of PHU increased the antioxidant capacity of plasma from rats. The ability of PHU to scavenge free radicals suggests that this drug may be potentially useful in counteracting free radical-mediated diseases.

Resonance. The missing phenomenon in hemodynamics.

To simulate a short segment of the aorta, we studied wave propagation in an elastic tube with a side branch balloon. The small balloon simulated the organ (group of arterioles). Ligation of this side branch would reduce the moduli of the higher harmonics when the length of the side branch was appropriate. Electrical analogy of vessels was used to analyze this phenomenon. This simulation can explain the ligation results we found in rats. It may also clarify the discrepancies between the prediction of the Womersley equation and the experimental results. We suggest that the aorta and the closely attached organ can produce coupled oscillation; theoretically, this structure is equivalent to a resonance circuit.

The cervix-ripening effect of 15-methyl-prostaglandin F2 alpha methyl ester before vacuum aspiration for termination of early pregnancy in primigravidae.

A double blind clinical trial on prostaglandin for cervix ripening prior to vacuum aspiration for termination of early pregnancy in primigravidae was carried out in 68 patients with 6-11 week's gestation. The patients were randomly treated with either 1.0 mg of 15-methyl-prostaglandin F2 alpha methyl ester vaginal suppository or a placebo suppository. A mean cervical dilation to 6.6 mm was achieved with minimal side effects in the prostaglandin group as against 4.3 mm in the placebo group. The 15-methyl-prostaglandin F2 alpha methyl ester suppository technique seems to be effective, safe and simple in ripening the cervix for termination of early pregnancy in primigravidae.

Hormone changes during the menstrual cycle of Chinese women.

The concentrations of LH, FSH, prolactin, oestradiol and progesterone in serum were measured daily during the menstrual cycle of 100 normal Chinese women. The cyclic changes in LH, FSH, oestradiol and progesterone were typical of ovulatory cycles in women of other ethnic groups as reported in the literature. The geometric mean of the LH midcycle peak value was 51 X 64 i.u./l, the FSH mid-cycle peak value was 11 X 52 i.u./l, the preovulatory oestradiol peak was 1229 X 12 pmol/l, and the progesterone luteal maximum was 53 X 27 nmol/l. The cyclic changes of prolactin concentrations were irregular: the value at mid-cycle was significantly higher than that at the follicular or luteal phases. A correlation between the length of the cycle and mean concentrations of LH and oestradiol at different stages throughout the cycle was shown.

Prostaglandin E and F2 alpha levels in plasma and amniotic fluid during mid-trimester abortion induced by trichosanthin.

This paper gives a brief account concerning the estimation of PGE and PGF2 alpha concentrations in both plasma and amniotic fluid by radioimmunoassay in mid-trimester abortion induced by Trichosanthin. The significant change in amniotic fluids as compared with that in plasma is brought out for discussion as an aid to the study of mechanism of abortion by Trichosanthin.

PIP: Trichosanthin, an abortifacient plant protein isolated from Radix Trichosanthes, has a success rate of approximately 98%. This paper examines changes in prostaglandin (PG) E and F2alpha levels in both plasma and amniotic fluid by radioimmunoassay in midtrimester abortion induced by Trichosanthin. Trichosanthin was given by transabdominal amniotic injection (average dosage, 8-11 mg) in 8 women (7 primigravidae and 1 secundigravida) of 17-24 weeks gestation. Venous blood and amniotic fluid samples were collected both before and after drug administration. PGE and F2alpha in plasma and amniotic fluid were measured by radioimmunoassay based upon a method described by the Pekin Institute of Zoology. Injection-abortion time ranged from 72-168 hours with a mean of 100 hours. No significant change was observed between concentrations of PGE and PGF2alpha in the plasma before and after drug administration, probably due to the fact that the 2 PGs were released in pulsatile fashion instead of continuous, and because PGs have short half-life in the circulating blood. In contrast, both PGE and PGF2alpha levels in the amniotic fluid were markedly increased after Trichosanthin administration, the increments amounting to 12.8 and 23.8 times the preinjection value respectively (p0.001). In midtrimester abortion, Trichosanthin exerts a selective action on the syncytiotrophoblasts as demonstrated by their degenerative and necrotic changes and simultaneous deciduitis. These injuries are followed by impairment of the endocrine functions of the placenta, a disintegration of lysosomes inside the decidual cells, and a decreased stability of the lysosomal membrane, an increase of synthesis of PGE and PGF2alpha both in the fetal membrane and decidua, and finally, complete abortion.