Basal lamina of human seminiferous tubule—Its role in material transport

Summary Electron microscopy of testicular biopsies obtained from two adult males with tunica vaginal hydrocele revealed some protrusions from the basal lamina to the germinal epithelium in the seminiferous tubule. The protrusions were of three types: some between the spermatogonia and Sertoli cells, some directly within the Sertoli cell cytoplasm and others inside the spermatogonia. The protrusions inside the spermatogonia were only 0.5 m deep whereas the other types were from 1–11 m deep. Occasionally some cut off portions of these protrusions were seen inside the ground cytoplasm of the Sertoli cell without an apparent connection with the original stalk. The matrix of the protrusions contained a homogenous component (composed of a fine filamentous element) and granular and membranous components. These components closely resemble the materials found in the basal lamina of the seminiferous tubule. It has been suggested that under mild pathological conditions, i.e., hydrocele, the junctions between the seminiferous tubule epithelium and the basal lamina become somewhat more flexible. As a result, the protrusions become longer and a passage might be formed to allow the flow of raw materials in or out of the seminiferous tubule.This work was supported by USPHS Research Grant HD-03266 and GRS 94802     

Outbreak of mumps virus genotype G infection in tribal individuals during 2016–17 in India

Tribal individuals presented with fever and uni‐ or bi‐lateral parotitis in Galonda and Silli villages (Dadra and Nagar Haveli, India) between 2 October 2016 and 19 March 2017. Consequently, the magnitude and epidemiological characteristics of the outbreak were investigated. Overall, 139 cases of suspected mumps were identified in both the above villages. Most of the suspected cases were 5–15 years old, the exceptions being three adults who had no noticeable complications. Specimens were collected from 42 of the suspected cases and their close contacts (n = 39) for laboratory investigation. Mumps infection was laboratory‐confirmed in 73.8% and 20.5% of the suspected cases and contacts, respectively. Mumps was confirmed in seven adults aged 17–42 years, including three suspected cases and four contacts. To the best of our knowledge, this is the first report of a complete virus genome circulating among tribal individuals. Sequencing and phylogenetic studies revealed circulation of mumps virus genotype G in these tribal villages with 99% identity to a mumps virus detected in the UK (1996) and Canada (2009). Comparison with Indian mumps viruses revealed 99% and 98% identity to previously reported isolates from Pune during 2012 and 1986, respectively. Although the outbreak was large, no major complications were reported in the tribal villages. Detection of asymptomatic mumps in numerous close contacts indicates the importance of laboratory investigations in an outbreak setting.

     

Discovery of 5-(2-chloro-4′-(1H-imidazol-1-yl)-[1,1′-biphenyl]-4-yl)-1H-tetrazole as potent and orally efficacious S-nitrosoglutathione reductase (GSNOR) inhibitors for the potential treatment of COPD

Endogenous nitrosothiols (SNOs) including S-nitrosoglutathione (GSNO) serve as reservoir for bioavailable nitric oxide (NO) and mediate NO-based signaling, inflammatory status and smooth muscle function in the lung. GSNOR inhibition increases pulmonary GSNO and induces bronchodilation while reducing inflammation in lung diseases. In this letter, design, synthesis and structure–activity relationships (SAR) of novel imidazole-biaryl-tetrazole based GSNOR inhibitors are described. Many potent inhibitors (30, 39, 41, 42, 44, 45 and 58) were identified with low nanomolar activity (IC₅₀s: <15?nM) along with adequate metabolic stability. Lead compounds 30 and 58 exhibited good exposure and oral bioavailability in mouse pharmacokinetic (PK) study. Compound 30 was selected for further profiling and revealed comparable mouse and rat GSNOR potency, high selectivity against alcohol dehydrogenase (ADH) and carbonyl reductase (CBR1) family of enzymes, low efflux ratio and permeability in PAMPA, a high permeability in CALU-3 assay, significantly low hERG activity and minimal off-target activity. Further, an in vivo efficacy of compound 30 is disclosed in cigarette smoke (CS) induced mouse model for COPD.     

Fatal <Emphasis Type="Italic">Talaromyces marneffei</Emphasis> Infection in a Patient with Autoimmune Hepatitis

Talaromyces marneffei, previously known as Penicillium marneffei, is the most important pathogenic thermally dimorphic fungus causing systemic mycosis in Southeast Asia. Traditionally, T. marneffei infection in human was mainly associated with acquired immunodeficiency syndrome caused by HIV infection. In recent years, there has been an increasing number of T. marneffei infections reported in non-HIV-infected patients with other immunocompromised conditions, including autoantibodies against interferon-gamma, systemic lupus erythematosis, solid organ transplantation, Job’s syndrome, hematological malignancies, and use of novel targeted therapies. In this article, we describe the first case of fatal T. marneffei infection in a patient with underlying autoimmune hepatitis, presented as fever without localizing features. The diagnosis of talaromycosis was confirmed with the identification of the fungi isolated from the blood culture specimen by conventional methods and using matrix-assisted laser desorption–ionization time-of-flight mass spectrometer. This case shows the importance of a high index of suspicion, particularly for such a highly fatal but potentially treatable fungal infection.     

Integrated Signaling in Flower Senescence: An Overview

Flower senescence is the terminal phase of developmental processes that lead to the death of flower, which include, flower wilting, shedding of flower parts and fading of blossoms. Since it is a rapid process as compared to the senescence of other parts of the plant it therefore provides excellent model system for the study of senescence. During flower senescence, developmental and environmental stimuli enhance the upregulation of catabolic processes causing breakdown and remobilization of cellular constituents. Ethylene is well known to play regulatory role in ethylene-sensitive flowers while in ethylene-insensitive flowers abscisic acid (ABA) is thought to be primary regulator. Subsequent to perception of flower senescence signal, death of petals is accompanied by the loss of membrane permeability, increase in oxidative and decreased level of protective enzymes. The last stages of senescence involve the loss of of nucleic acids (DNA and RNA), proteins and organelles, which is achieved by activation of several nucleases, proteases and wall modifiers. Environmental stimuli such as pollination, drought and other stresses also affect senescence by hormonal imbalance. In this article we have covered the following: perception mechanism and specificity of flower senescence, flower senescence-associated events, like degradation of cell membranes, proteins and nucleic acids, environmental/external factors affecting senescence, like pollination and abiotic stress, hormonal and non-hormonal regulation of flower/petal senescence and finally the senescence associated genes (SAGs) have also been described.Key Words: environmental factors, ethylene, flowers, petals, plant hormones, pollination, programmed cell death, senescence, senescence-associated genes     

Post-translational formation of hypusine in eIF5A: implications in human neurodevelopment

Hypusine [N^ε-(4-amino-2-hydroxybutyl)lysine] is a derivative of lysine that is formed post-translationally in the eukaryotic initiation factor 5A (eIF5A). Its occurrence at a single site in one cellular protein defines hypusine synthesis as one of the most specific post-translational modifications. Synthesis of hypusine involves two enzymatic steps: first, deoxyhypusine synthase (DHPS) cleaves the 4-aminobutyl moiety of spermidine and transfers it to the ε-amino group of a specific lysine residue of the eIF5A precursor protein to form an intermediate, deoxyhypusine [N^ε-(4-aminobutyl)lysine]. This intermediate is subsequently hydroxylated by deoxyhypusine hydroxylase (DOHH) to form hypusine in eIF5A. eIF5A, DHPS, and DOHH are highly conserved in all eukaryotes, and both enzymes exhibit a strict specificity toward eIF5A substrates. eIF5A promotes translation elongation globally by alleviating ribosome stalling and it also facilitates translation termination. Hypusine is required for the activity of eIF5A, mammalian cell proliferation, and animal development. Homozygous knockout of any of the three genes, Eif5a, Dhps, or Dohh, leads to embryonic lethality in mice. eIF5A has been implicated in various human pathological conditions. A recent genetic study reveals that heterozygous germline EIF5A variants cause Faundes–Banka syndrome, a craniofacial–neurodevelopmental malformations in humans. Biallelic variants of DHPS were identified as the genetic basis underlying a rare inherited neurodevelopmental disorder. Furthermore, biallelic DOHH variants also appear to be associated with neurodevelopmental disorder. The clinical phenotypes of these patients include intellectual disability, developmental delay, seizures, microcephaly, growth impairment, and/or facial dysmorphisms. Taken together, these findings underscore the importance of eIF5A and the hypusine modification pathway in neurodevelopment in humans.

     

Rab6 regulates both ZW10/RINT-1 and conserved oligomeric Golgi complex-dependent Golgi trafficking and homeostasis

We used multiple approaches to investigate the role of Rab6 relative to Zeste White 10 (ZW10), a mitotic checkpoint protein implicated in Golgi/endoplasmic reticulum (ER) trafficking/transport, and conserved oligomeric Golgi (COG) complex, a putative tether in retrograde, intra-Golgi trafficking. ZW10 depletion resulted in a central, disconnected cluster of Golgi elements and inhibition of ERGIC53 and Golgi enzyme recycling to ER. Small interfering RNA (siRNA) against RINT-1, a protein linker between ZW10 and the ER soluble N-ethylmaleimide-sensitive factor attachment protein receptor, syntaxin 18, produced similar Golgi disruption. COG3 depletion fragmented the Golgi and produced vesicles; vesicle formation was unaffected by codepletion of ZW10 along with COG, suggesting ZW10 and COG act separately. Rab6 depletion did not significantly affect Golgi ribbon organization. Epistatic depletion of Rab6 inhibited the Golgi-disruptive effects of ZW10/RINT-1 siRNA or COG inactivation by siRNA or antibodies. Dominant-negative expression of guanosine diphosphate-Rab6 suppressed ZW10 knockdown induced-Golgi disruption. No cross-talk was observed between Rab6 and endosomal Rab5, and Rab6 depletion failed to suppress p115 (anterograde tether) knockdown-induced Golgi disruption. Dominant-negative expression of a C-terminal fragment of Bicaudal D, a linker between Rab6 and dynactin/dynein, suppressed ZW10, but not COG, knockdown-induced Golgi disruption. We conclude that Rab6 regulates distinct Golgi trafficking pathways involving two separate protein complexes: ZW10/RINT-1 and COG.     

CFD Analysis of Pulsatile Flow and Non-Newtonian Behavior of Blood in Arteries

CFD analysis plays an important role in the area of analysis of blood flow as in-vivo measurements of blood flow is costly and easily not accessible. This paper presents simulation of blood flow in healthy and stenosed coronary artery 2- D models. The simulation was done considering non-Newtonian behavior of blood and pulsatile nature of blood flow which is close to physical scenario. Pressure distribution, velocity distribution and wall shear were examined to understand their effect on Atherosclerosis.     

Heightened Delta Power during Slow-Wave-Sleep in Patients with Rett Syndrome Associated with Poor Sleep Efficiency

Sleep problems are commonly reported in Rett syndrome (RTT); however the electroencephalographic (EEG) biomarkers underlying sleep dysfunction are poorly understood. The aim of this study was to analyze the temporal evolution of quantitative EEG (qEEG) biomarkers in overnight EEGs recorded from girls (2–9 yrs. old) diagnosed with RTT using a non-traditional automated protocol. In this study, EEG spectral analysis identified high delta power cycles representing slow wave sleep (SWS) in 8–9h overnight sleep EEGs from the frontal, central and occipital leads (AP axis), comparing age-matched girls with and without RTT. Automated algorithms quantitated the area under the curve (AUC) within identified SWS cycles for each spectral frequency wave form. Both age-matched RTT and control EEGs showed similar increasing trends for recorded delta wave power in the EEG leads along the antero-posterior (AP). RTT EEGs had significantly fewer numbers of SWS sleep cycles; therefore, the overall time spent in SWS was also significantly lower in RTT. In contrast, the AUC for delta power within each SWS cycle was significantly heightened in RTT and remained heightened over consecutive cycles unlike control EEGs that showed an overnight decrement of delta power in consecutive cycles. Gamma wave power associated with these SWS cycles was similar to controls. However, the negative correlation of gamma power with age (r = -.59; p<0.01) detected in controls (2–5 yrs. vs. 6–9 yrs.) was lost in RTT. Poor % SWS (i.e., time spent in SWS overnight) in RTT was also driven by the younger age-group. Incidence of seizures in RTT was associated with significantly lower number of SWS cycles. Therefore, qEEG biomarkers of SWS in RTT evolved temporally and correlated significantly with clinical severity.     

MiR-101 Induces Senescence and Prevents Apoptosis in the Background of DNA Damage in MCF7 Cells

Moderately increased DNA damage due to the exogenous miR-101 (4 fold) over-expression in MCF7 cells was substantiated by an increase in the number of γ-H2AX foci, correlating with a simple-to-do Halo-assay. miR-101 induced mild/moderate DNA damage favoured senescence rather than apoptosis. An experimental support emanated from the induced mild/moderate DNA damage with 1 µM/5 µM etoposide in MCF7 cells, which resulted in an endogenous miR-101 over-expression (10/4 fold, respectively), followed by senescence. On the other hand, the severe DNA damage induced with 10 µM etoposide, resulted in a low (<1 fold) endogenous expression of miR-101 and an elevated percentage of apoptotic cells. Using bioinformatics tools along with in-vitro and in-vivo validations, miR-101 was found to target and downregulate the mRNA expression of UBE2N and SMARCA4, involved in DNA damage repair (DDR) pathways. Recovery of the expression of the two novel targets in anti-miR-101 transfection validated the results. We conclude that a threshold range of over-expressed miR-101, capable of inducing mild/moderate DNA damage, is sensed by cells to become senescent. The observation derives further support from in-silico protein-protein network analysis where the two novel targets showed their involvement in senescence pathway.