The mitochondrial RNA granule modulates manganese-dependent cell toxicity.

Prolonged manganese exposure causes manganism, a neurodegenerative movement disorder. The identity of adaptive and nonadaptive cellular processes targeted by manganese remains mostly unexplored. Here we study mechanisms engaged by manganese in genetic cellular models known to increase susceptibility to manganese exposure, the plasma membrane manganese efflux transporter SLC30A10 and the mitochondrial Parkinson's gene PARK2. We found that SLC30A10 and PARK2 mutations as well as manganese exposure compromised the mitochondrial RNA granule composition and function, resulting in disruption of mitochondrial transcript processing. These RNA granule defects led to impaired assembly and function of the mitochondrial respiratory chain. Notably, cells that survived a cytotoxic manganese challenge had impaired RNA granule function, thus suggesting that this granule phenotype was adaptive. CRISPR gene editing of subunits of the mitochondrial RNA granule, FASTKD2 or DHX30, as well as pharmacological inhibition of mitochondrial transcription-translation, were protective rather than deleterious for survival of cells acutely exposed to manganese. Similarly, adult Drosophila mutants with defects in the mitochondrial RNA granule component scully were safeguarded from manganese-induced mortality. We conclude that impairment of the mitochondrial RNA granule function is a protective mechanism for acute manganese toxicity.

Temporomandibular Disorder: a practical guide for dental practitioners in diagnosis and management.

Temporomandibular disorder is a broad term encompassing pain and/or dysfunction of the masticatory musculature and the temporomandibular joints. Pain arising from a temporomandibular disorder is a common reason for seeking dental care. It is essential that dental practitioners are able to accurately diagnose and manage this condition. Identifying people at highest risk of developing a temporomandibular disorder and knowing which procedures are more likely to initiate or exacerbate a temporomandibular disorder, are important to reduce the likelihood of its acute and chronic presentation. The aim of this paper is to provide the dental practitioner with a clinical guideline for reference including practical tools to examine, diagnose and manage patients with temporomandibular disorder. In addition the risk profile of patients and procedures is explored to help minimize the occurrence of temporomandibular disorders and mitigate its symptoms. The scope of the dental practitioner in the management of acute and chronic temporomandibular disorders is presented, with guidance about when referral to a specialist is indicated.

Neurodevelopmental disorders: mechanisms and boundary definitions from genomes, interactomes and proteomes.

Neurodevelopmental disorders such as intellectual disability, autism spectrum disorder and schizophrenia lack precise boundaries in their clinical definitions, epidemiology, genetics and protein-protein interactomes. This calls into question the appropriateness of current categorical disease concepts. Recently, there has been a rising tide to reformulate neurodevelopmental nosological entities from biology upward. To facilitate this developing trend, we propose that identification of unique proteomic signatures that can be strongly associated with patient's risk alleles and proteome-interactome-guided exploration of patient genomes could define biological mechanisms necessary to reformulate disorder definitions.

The WASH complex, an endosomal Arp2/3 activator, interacts with the Hermansky-Pudlak syndrome complex BLOC-1 and its cargo phosphatidylinositol-4-kinase type IIα.

Vesicle biogenesis machinery components such as coat proteins can interact with the actin cytoskeleton for cargo sorting into multiple pathways. It is unknown, however, whether these interactions are a general requirement for the diverse endosome traffic routes. In this study, we identify actin cytoskeleton regulators as previously unrecognized interactors of complexes associated with the Hermansky-Pudlak syndrome. Two complexes mutated in the Hermansky-Pudlak syndrome, adaptor protein complex-3 and biogenesis of lysosome-related organelles complex-1 (BLOC-1), interact with and are regulated by the lipid kinase phosphatidylinositol-4-kinase type IIα (PI4KIIα). We therefore hypothesized that PI4KIIα interacts with novel regulators of these complexes. To test this hypothesis, we immunoaffinity purified PI4KIIα from isotope-labeled cell lysates to quantitatively identify interactors. Strikingly, PI4KIIα isolation preferentially coenriched proteins that regulate the actin cytoskeleton, including guanine exchange factors for Rho family GTPases such as RhoGEF1 and several subunits of the WASH complex. We biochemically confirmed several of these PI4KIIα interactions. Of importance, BLOC-1 complex, WASH complex, RhoGEF1, or PI4KIIα depletions altered the content and/or subcellular distribution of the BLOC-1-sensitive cargoes PI4KIIα, ATP7A, and VAMP7. We conclude that the Hermansky-Pudlak syndrome complex BLOC-1 and its cargo PI4KIIα interact with regulators of the actin cytoskeleton.

Duplication of urethra: review of three cases.

Urethral duplication is a rare congenital anomaly and presents as a spectrum of varying severity. This report present three cases of duplication of urethra whose presentation ranged from prepubic sinus to a complete urethral duplication. They were investigated using ultrasonography, retrograde urethrography and cystoscopy. They were treated depending on the severity of the anomaly. The management of these cases is discussed and the current literature on this rare entity reviewed.

Silver nitrate sclerotherapy for chyluria: evaluation for the optimal instillation regime.

OBJECTIVE: To compare the efficacy of the 3-instillation regime with the 9-instillation regime of 1% silver nitrate solution for renal pelvic instillation sclerotherapy in patients with chyluria. PATIENTS AND METHODS: 59 Patients with chyluria were prospectively evaluated and randomized into two groups. The study group received 3 instillations (n = 29) while the control group received 9 instillations (n = 30). 1% silver nitrate was used. RESULTS: In the study group of the 29 patients given sclerotherapy, 28 (96.5%) had complete symptomatic relief while symptoms persisted in 1 (3.44%). None of the patients had hematuria or any other severe complication. In the control group of the 30 patients given sclerotherapy, 29 (96.66%) had complete symptomatic relief while chyluria persisted in 1 (3.33%), 5 patients developed hematuria (16.66%) and 1 patient developed fever with UTI (3.33%). The results of the 3-instillation regime match the results of the more commonly used 9-instillation regime. With this shorter regime, the hospital stay, morbidity and overall complication rate is less and it is more cost effective. CONCLUSION: We recommend the 3-instillation regimen in patients with chyluria refractory to conservative treatment.

Clinical and dosimetric factors associated with a prolonged feeding tube requirement in patients treated with chemoradiotherapy (CRT) for head and neck cancers.

BACKGROUND: Patients treated with chemoradiotherapy (CRT) for head and neck cancers often require feeding tubes (FTs) due to toxicity. We sought to identify factors associated with a prolonged FT requirement. PATIENTS AND METHODS: We retrospectively reviewed 80 patients treated with CRT for head and neck cancers. The pharyngeal constrictors (PCs), supraglottic larynx (SGL), and glottic larynx (GL) were contoured and the mean radiation doses and the volumes of each receiving >40, 50, 60, and 70 Gy (V40, V50, V60, and V70) were determined. RESULTS: A total of 33 of 80 patients required a FT either before or during the course of CRT. Fifteen patients required the FT for > or = 6 months. On univariate analysis, significant factors associated with a prolonged FT requirement were mean PC dose, PC-V60, PC-V70, SGL dose, SGL-V70, and advanced T3-T4 disease. Multivariate analyses found both PC-V70 and T3-T4 disease as significant factors .The proportions of patients requiring a FT > or = 6 months were 8% and 28% for treatment plans with PC-V70 <30% and > or = 30%, respectively. CONCLUSIONS: Increased radiation dose to the PCs is associated with a higher risk of a prolonged FT need. Dose sparing of the PC muscles may reduce this risk.

Design-based estimation of surface area in thick tissue sections of arbitrary orientation using virtual cycloids.

Surface area is a first-order stereological parameter with important biological applications, particularly at the intersection of biological phases. To deal with the inherent anisotropy of biological surfaces, state-of-the-art design-based methods require tissue rotation around at least one axis prior to sectioning. This paper describes the use of virtual cycloids for surface area estimation of objects and regions in thick, transparent tissue sections cut at any arbitrary (convenient) orientation. Based on the vertical section approach of Baddeley et al., the present approach specifies the vertical axis as the direction of sectioning (i.e. the direction perpendicular to the tissue section), and applies computer-generated cycloids (virtual cycloids) with their minor axis parallel to the vertical axis. The number of surface-cycloid intersections counted on focal planes scanned through the z-axis is proportional to the surface area of interest in the tissue, with no further assumptions about size, shape or orientation. Optimal efficiency at each x-y location can be achieved by three virtual cycloids orientated with their major axes (which are parallel to the observation planes) mutually at an angle of 120 degrees. The major practical advantage of the present approach is that estimates of total surface area (S) and surface density (SV) can be obtained in tissue sections cut at any convenient orientation through the reference space.

Studies on the immunomodulatory activity of flavonoidal fraction of Tephrosia purpurea.

The flavonoid fraction of Tephrosia purpurea (FFTP) was studied for its effect on cellular and humoral functions and on macrophage phagocytosis in mice. Oral administration of FFTP (10-40 mg/kg) significantly inhibited sheep red blood cells (SRBC)-induced delayed-type hypersensitivity reactions. It also produced a significant, dose-related decrease in sheep erythrocyte-specific haemagglutination antibody titre. However, the fraction failed to show a significant change in the macrophage phagocytic activity. The results obtained indicate the ability of the flavonoidal fraction of T. purpurea to modulate both the cell-mediated and the humoral components of the immune system.

Investigations into the immunomodulatory activity of Argyreia speciosa.

The objective of the present study was to investigate the immunomodulatory activity of Argyreia speciosa on cellular and humoral immunity. Oral administration of the ethanolic extract of A. speciosa root (ASEE), at the doses of 50, 100 and 200 mg/kg in mice, dose-dependently potentiated the delayed-type hypersensitivity reaction induced both by sheep red blood cells (SRBC) and oxazolone. It significantly enhanced the production of circulating antibody titre in mice in response to SRBC. ASEE failed to show any effect on macrophage phagocytosis. Chronic administration of ASEE significantly ameliorated the total white blood cell count and also restored the myelosuppressive effects induced by cyclophosphamide. The present investigation reveals that ASEE possesses immunomodulatory activity.

Preliminary evaluation of anti-inflammatory and anti-arthritic activity of S. lappa, A. speciosa and A. aspera.

Saussurea lappa, Argyreia speciosa and Achyranthes aspera are well known Indian medicinal plants used in the indigenous systems of medicine for the treatment of inflammatory conditions. The ethanolic extracts of the plants at the doses of 50, 100 and 200 mg/kg, p.o. were screened for their effect on acute and chronic inflammation induced in mice and rats. S. lappa and A. speciosa were found to significantly inhibit paw edema induced by carrageenan and Freund's complete adjuvant and to prevent accumulation of inflammatory cells in carrageenan-induced peritonitis at doses of 50-200 mg/kg. A. aspera inhibited these inflammatory responses at doses of 100-200 mg/kg. The studies reveal that the ethanolic extracts of S. lappa, A. speciosa and A. aspera possess anti-inflammatory and anti-arthritic activity and support the rationale behind the traditional use of these plants in inflammatory conditions.

Stereological length estimation using spherical probes.

Lineal structures in biological tissue support a wide variety of physiological functions, including membrane stabilization, vascular perfusion, and cell-to-cell communication. In 1953, Smith and Guttman demonstrated a stereological method to estimate the total length density (Lv) of linear objects based on random intersections with a two-dimensional sampling probe. Several methods have been developed to ensure the required isotropy of object-probe intersections, including isotropic-uniform-random (IUR) sections, vertical-uniform-random (VUR) slices, and isotropic virtual planes. The disadvantages of these methods are the requirements for inconvenient section orientations (IUR, VUR) or complex counting rules at multiple focal planes (isotropic virtual planes). To overcome these limitations we report a convenient and straightforward approach to estimate Lv and total length, L, for linear objects on tissue sections cut at any arbitrary orientation. The approach presented here uses spherical probes that are inherently isotropic, combined with unbiased fractionator sampling, to demonstrate total L estimation for thin nerve fibres in dorsal hippocampus of the mouse brain.

Environmental initiative prioritization with a Delphi approach: a case study.

India is fast finding its place in the industrialized world and that is beginning to raise its environmental consciousness. The Delphi technique was used to prioritize specific needs and articulate a sustainable urban improvement strategy for the city of Mumbai (formerly Bombay). The Delphi technique is a means of achieving consensual validity among raters by providing them feedback regarding other raters' responses. Mumbai has several indigenous environmental groups that were tapped for activists and leaders; the study was conducted using ten environmentalists. In the initial phases the responses resulted in a range of possible program alternatives. The last two stages helped to seek out information that generated a consensus on the part of the respondent group. Statistical analysis methods included a hierarchical cluster analysis, mean, median, mode, and percent of agreement calculations using SPSS software. The face-to-face discussion in phase 4 clarified some issues and helped the group as a whole to outline the strategy for putting in place the essential elements of a framework to improve the quality of life in an urban environment.

Surgical salvage after chemoradiation of head and neck cancer: complications and outcomes.

The treatment of advanced squamous cell carcinoma involves a multidisciplinary approach among various physicians and ancillary personnel. The role of the head and neck surgeon continues to change, as concurrent chemotherapy and radiotherapy protocols have evolved in the initial management of this challenging patient population. More and more, the surgeon is called upon to operate on those patients with persistent or recurrent disease despite initial treatment with chemotherapy and radiotherapy. The purpose of this article is to analyze complications and outcomes of surgery in patients who have already received radiotherapy and chemotherapy.

Bias of a length density estimator based on vertical projections.

Attention is paid to the stereological estimator of the length density of lineal features in three-dimensional space. In Gokhale (J. Microsc. (1990) 159, 133-141), the estimator based on measurements performed on a projection of the content of vertical slices with a given thickness Delta was derived. The aim of this paper is to show that using five vertical slices with systematically chosen orientations yields a reliable result, i.e. the bias of the estimator is smaller than 5%. In the case of a choice of the vertical axis such that most lineal features are not perpendicular or nearly perpendicular to the vertical axis, a reliable result can be obtained using only three systematic orientations of vertical slices.

Efficient estimation of number density in opaque material microstructures: the large-area disector.

Unbiased and efficient estimation of number density of features in opaque material microstructures has been quite difficult. In this contribution a montage-based efficient serial sectioning technique is presented as a practical solution for efficient estimation of number density in such microstructures. The new technique utilizes a combination of digital image analysis and unbiased disector sampling procedures.

Influence of ethanolic extract of Tephrosia purpurea Linn. on mast cells and erythrocytes membrane integrity.

The ethanolic extract of T. purpurea Linn. was studied for its in vitro effect on rat mast cell degranulation and erythrocyte membrane integrity in vitro. The extract in concentration of 25-200 microg/ml showed a dose-dependant inhibition of rat mast cell degranulation induded by compound 48/80 and egg albumin. T. purpurea extract was found to inhibit haemolysis of erythrocytes induced by hypotonic solution but accelerated haemolysis induced by heat at a concentration of 100 microg/ml. The studies reveal that the ethanolic extract of T. purpurea may inhibit degranulation of mast cells by a mechanism other than membrane stabilization.