A novel gelatinized barium sulfate injection method for assessment of bronchoalveolar lavage parameters.

INTRODUCTION: Bronchoalveolar lavage (BAL) is frequently used in pulmonary medicine though it requires further optimization. Practical obstacles such as patient safety and procedural limitation have to date precluded large, controlled trials aimed at standardization of BAL procedure. Indeed, BAL guidelines are based on observational data. Innovative research methods are necessary to advance the clinical practice of BAL. METHODS: In our study, we evaluated the effect of injecting a gelatinized barium solution into different lobes and segments of cadaveric lungs. As the technique requires an irreversible injection into lung airspaces, it is not suitable for in vivo purposes. We measured the volume returned from BAL as well as the distribution of BAL injection via dissection. Segmental anatomic orientation was compared to a radiologist's impression of plain film radiographs taken of injected lungs. RESULTS: Mean injected volume distributions were greatest in the upper lobes and lowest in the lower lobes; mean ratios of injected volume distribution to lung lobe volume also followed this trend. Cannulated bronchi orders favored lower branches in the upper lobe and higher branches in the lower lobes. Segmental anatomy varied by the lung lobe injected and was most varied in the lower lobes. CONCLUSION: This novel gelatinized-barium injection technique provides a minimally complex method to yield clinically meaningful feedback on the performance of BAL. The technique is also adaptable to study of procedural parameters in the context of variable lung anatomies and pathologies.

Bioinformatics analysis of structural protein to approach a vaccine candidate against Vibrio cholerae infection.

The bacteria Vibrio cholerae causes cholera, an acute diarrheal infection that can lead to dehydration and even death. Over 100,000 people die each year as a result of epidemic diseases; vaccination has emerged as a successful strategy for combating cholera. This study uses bioinformatics tools to create a multi-epitope vaccine against cholera infection using five structural polyproteins from the V. cholerae (CTB, TCPA, TCPF, OMPU, and OMPW). The antigenic retrieved protein sequence were analyzed using BCPred and IEDB bioinformatics tools to predict B cell and T cell epitopes, respectively, which were then linked with flexible linkers together with an adjuvant to boost it immunogenicity. The construct has a theoretical PI of 6.09, a molecular weight of 53.85 kDa, and an estimated half-life for mammalian reticulocytes in vitro of 4.4 h. These results demonstrate the construct's longevity. The vaccine design was docked against the human toll-like receptor (TLR) to evaluate compatibility and effectiveness; also other additional post-vaccination assessments were carried out on the designed vaccine. Through in silico cloning, its expression was determined. The results show that it has a CAI value of 0.1 and GC contents of 58.97% which established the adequate expression and downstream processing of the vaccine construct, and our research demonstrated that the multi-epitope subunit vaccine exhibits antigenic characteristics. Additionally, we carried out an in silico immunological simulation to examine the immune reaction to an injection. Our results strongly suggest that the vaccine candidate on further validation would induce immune response against the V. cholerae infection.

Immunoinformatics design of multi-epitope peptide for the diagnosis of Schistosoma haematobium infection.

Schistosoma haematobium has been identified as a significant cause of urogenital disease, as well as a risk factor for bladder cancer and HIV/AIDS. The parasites are obtained trans-dermally by swimming or wading in contaminated freshwater, and they are also transmitted to humans by freshwater snails. The organisms infect the vasculature of the gastrointestinal or genitourinary tracts. Worms live in blood vessels and lay eggs that become embedded in the bladder wall, causing chronic immune-mediated disease and squamous cell carcinoma growth. The primary goal of this research is to predict and design a novel synthetic protein containing multiple immunodominant B cell epitopes using three schistosome proteins: XP-012801068.2, XP-012801892.2, and XP-012793835.2 softwares were used to analyze the proteins' primary, secondary, and tertiary structures (BepiPred, BcPred).The B cell construct was then evaluated using I-TASSER server, and physicochemical properties, as well as homology modeling of the 3 D structure of the protein, was obtained. In silico analyses revealed regions with high immunogenicity. For XP-012801068.2, three epitopes are found between residues 292-334, 3-22, and 314-333; for XP-012801892.2, three epitopes are found in the residues 184-236, 81-100, and 329-348 for XP-012793835.2, four epitopes are found in the residues 185-222, 469-512, 649-713, and 338-357. The construct's has an average length of 308 bp, instability index of 49.96, theoretical PI of 4.2 and a C score -1.59. Furthermore, these parameters analyzed reveals that the constructed multi-epitope peptide has the potential to provide a theoretical basis for the development of a Schistosoma haematobium diagnostic kit.Communicated by Ramaswamy H. Sarma.

A primary solitary vascular tumor of calcaneum: case report and review of literature.

Epithelioid hemangioendothelioma (EHE) is a rare vascular tumor with metastatic potential and estimated prevalence of less than one case per million. Among the musculoskeletal system, the long bones are commonly involved with approximately half patients experiencing multicentric involvement. Clinical course of EHE is often variable and nonspecific. Poorly demarcated osteolytic lesions are most commonly seen radiologically. Diagnostic confirmation is usually obtained by biopsy and histopathological exam, including immunostaining for endothelial markers. We present a rare case of unicentric EHE involving the calcaneum. Our patient had an indolent course of disease after surgical resection and no recurrence in seven years on clinical and radiological surveillance.

Enhancing flowering plant functional richness improves wild bee diversity in vineyard inter-rows in different floral kingdoms.

Wild bees are threatened by multiple interacting stressors, such as habitat loss, land use change, parasites, and pathogens. However, vineyards with vegetated inter-rows can offer high floral resources within viticultural landscapes and provide foraging and nesting habitats for wild bees. Here, we assess how vineyard management regimes (organic vs. conventional; inter-row vegetation management) and landscape composition determine the inter-row plant and wild bee assemblages, as well as how these variables relate to functional traits in 24 Austrian and 10 South African vineyards. Vineyards had either permanent vegetation cover in untilled inter-rows or temporary vegetation cover in infrequently tilled inter-rows. Proportion of seminatural habitats (e.g., fallows, grassland, field margins) and woody structures (e.g., woodlots, single trees, tree rows) were used as proxies for landscape composition and mapped within 500-m radius around the study vineyards. Organic vineyard management increased functional richness (FRic) of wild bees and flowering plants, with woody structures marginally increasing species richness and FRic of wild bees. Wild bee and floral traits were differently associated across the countries. In Austria, several bee traits (e.g., lecty, pollen collection type, proboscis length) were associated with flower color and symmetry, while in South African vineyards, only bees' proboscis length was positively correlated with floral traits characteristic of Asteraceae flowers (e.g., ray-disk morphology, yellow colors). Solitary bee species in Austria benefitted from infrequent tillage, while ground nesting species preferred inter-rows with undisturbed soils. Higher proportions of woody structures in surrounding landscapes resulted in less solitary and corbiculate bees in Austria, but more aboveground nesting species in South Africa. In both countries, associations between FRic of wild bees and flowering plants were positive both in organic and in conventional vineyards. We recommend the use of diverse cover crop seed mixtures to enhance plant flowering diversity in inter-rows, to increase wild bee richness in viticultural landscapes.

SATB2 and MDM2 Immunoexpression and Diagnostic Role in Primary Osteosarcomas of the Jaw.

Primary osteosarcomas of the jaw (OSJ) are rare, accounting for 6% of all osteosarcomas. This study aims to determine the value of SATB2 and MDM2 immunohistochemistry (IHC) in differentiating OSJ from other jawbone mimickers, such as benign fibro-osseous lesions (BFOLs) of the jaw or Ewing sarcoma of the jaw. Certain subsets of osteosarcoma harbor a supernumerary ring and/or giant marker chromosomes with amplification of the 12q13-15 region, including the murine double-minute type 2 (MDM2) and cyclin-dependent kinase 4 (CDK4) genes. Special AT-rich sequence-binding protein 2 (SATB2) is an immunophenotypic marker for osteoblastic differentiation. Cases of OSJ, BFOLs (ossifying fibroma and fibrous dysplasia) of the jaw, and Ewing sarcoma of the jaw were retrieved from the Departments of Oral Pathology and Oral Medicine, Faculty of Dentistry, Obafemi Awolowo University and Lagos State University College of Medicine, Nigeria. All OSJ retrieved showed histologic features of high-grade osteosarcoma. IHC for SATB2 (clone EP281) and MDM2 (clone IF2), as well as fluorescence in situ hybridization (FISH) for MDM2 amplification, were performed on all cases. SATB2 was expressed in a strong intensity and diffuse staining pattern in all cases (11 OSJ, including a small-cell variant, 7 ossifying fibromas, and 5 fibrous dysplasias) except in Ewing sarcoma, where it was negative in neoplastic cells. MDM2 was expressed in a weak to moderate intensity and scattered focal to limited diffuse staining pattern in 27% (3/11) of cases of OSJ and negative in all BFOLs and the Ewing sarcoma. MDM2 amplification was negative by FISH in interpretable cases. In conclusion, the three cases of high-grade OSJs that expressed MDM2 may have undergone transformation from a low-grade osteosarcoma (LGOS). SATB2 is not a dependable diagnostic marker to differentiate OSJ from BFOLs of the jaw; however, it could serve as a valuable diagnostic marker in differentiating the small-cell variant of OSJ from Ewing sarcoma of the jaw, while MDM2 may be a useful diagnostic marker in differentiating OSJ from BFOLs of the jaw, especially in the case of an LGOS or high-grade transformed osteosarcoma.

Vitamin K protects against 7,12-dimethylbenz(A)anthracene induced hepatotoxicity in Wistar rats.

Humans are daily exposed to 7,12-dimethylbenz(a)anthracene (DMBA), a well known polycyclic aromatic hydrocarbons (PAH). This study investigated the role of dietary intake of Vitamin K (VK), a polyphenolic compound, with potential antioxidative properties, against DMBA-induced hepatotoxicity. Sixty experimental animals (120-150 g) were divided into six groups (A-F): Control, DMBA (80 mg/kg bw) only, VK (0.00 g/10 kg) diet only, VK (7.5 g/10 kg) diet only, DMBA + VK (0.0 g/10 kg) diet and DMBA + VK (7.5 g/10 kg) diet. Single oral administration of DMBA (80 mg/kg body weight) to Wistar rats resulted in hepatic damage after 16 weeks. DMBA significantly (P < .05) decreased the activities of catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) and glutathione peroxidase (GPx). Levels of reduced glutathione (GSH) and Vitamin C were significantly decreased with increase in malondialdehyde (MDA) and nitric oxide (NO) levels in serum and liver. Aspartate aminotransaminase (AST), alanine aminotransaminase (ALT), γ-glutamyltransferase (GGT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) activities were significantly (P < .05) elevated in the serum but reduced in the liver of DMBA-administered group. Ingestion of 7.5 g/10 kg VK diet prevented the up regulations in inflammatory biomarkers (granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin 17A (IL-17A)) which elicited liver damaged in the DMBA-treated group. DMBA induced hepatic alterations in DMBA-treated group but was restored to near normal in VK (7.5 g/10 kg) diet group. These findings suggest the protective potential of increased dietary intake of vitamin K against DMBA-induced hepatic dysfunction.

BRAF V600E mutation-specific immunohistochemical analysis in ameloblastomas: a 44-patient cohort study from a single institution.

PURPOSE: The aim of this study is to investigate the presence and prevalence of BRAF V600E mutation in ameloblastomas using anti-BRAF V600E monoclonal antibody (VE1 clone) and to identify any clinicopathologic correlation with BRAF V600E mutation in ameloblastoma. MATERIALS AND METHODS: The pathology files of the Department of Oral Pathology and Oral Medicine, Faculty of Dentistry, Lagos State University College of Medicine, Lagos, Nigeria, were searched for the diagnosis of ameloblastoma from 2016 to 2020. Archived non-decalcified formalin-fixed paraffin-embedded tissue underwent immunohistochemistry using anti-BRAF V600E antibody at the University of Pittsburgh, Pennsylvania. Clinicopathologic data such as age at diagnosis, gender, jaw bone involved (mandible or maxilla), tumor location (anterior or posterior) and histologic subtype were collected. The clinicopathologic parameters were analyzed using Chi-square test and Fisher's exact test according to the BRAF status. RESULTS: Forty-four cases of ameloblastoma were retrieved. The male to female ratio was 1.32:1. The average age of patients at diagnosis was 33.3 years. Thirty-nine cases were located in the mandible and 5 cases in the maxilla. Only cases in the mandible were positive for anti-BRAF V600E antibody (n = 15/39; 38.5%). There was a significant correlation between BRAF V600E expression in mandibular tumors and histologic subtype (p = 0.02); however, no significance was observed for gender, age and tumor location. CONCLUSION: BRAF V600E mutation preferentially occurs in mandibular ameloblastomas, especially in non-plexiform ameloblastomas. These patients may benefit therapeutically from the use of BRAF inhibitors.

Asynchrony among insect pollinator groups and flowering plants with elevation.

Mountains influence species distribution through differing climate variables associated with increasing elevation. These factors determine species niche ranges and phenology. Although the distribution patterns of some specific insect groups relative to elevation have been determined, how differing environmental conditions across elevation zones differentially influence the phenology of various insect groups is largely unknown. This is important in this era of rapid climate change. We assess here how species composition and seasonal peaks in abundance among different insect pollinator groups and flowering plants differ across four floristically distinct elevation zones up a sentinel mountain subject to strong weather events. We sampled insect pollinators in four major groups (bees, wasps, beetles and flies) over two spring seasons. Pollinator species composition across all elevation zones tracks flowering plant species composition. In terms of abundance, beetles were the dominant group across the three lower zones, but declined greatly in the summit zone, where flies and bees were more abundant. Bee abundance peaked earlier than the other groups across all four elevation zones, where there were significant peaks in abundance. Bee abundance peaked earlier than flowering plants at the middle zone and slightly later than flowering plants at the base zone, suggesting a mismatch. We conclude that, while elevation shapes species distribution, it also differentially influences species phenology. This may be of great significance in long-term assessment of species distribution in sensitive mountain ecosystems.

A severe presentation of chronic non-infectious osteomyelitis associated with ulcerative colitis: a case report.

BACKGROUND: Chronic Non-Infectious Osteomyelitis (CNO) is a chronic, relapsing, self-limiting inflammation of the bone. Although it is rare, CNO has been associated with inflammatory bowel disease and frequently precedes the initial diagnosis. We present a case of CNO in a patient with known ulcerative colitis in clinical remission who presented with purulent multifocal joint effusions in the setting of elevated inflammatory markers and fever suspicious for bacterial osteomyelitis. CASE PRESENTATION: Our patient is a 12-year-old girl with ulcerative colitis who presented with fevers and insidious onset of joint pain at multiple sites. She had multiple joint effusions on imaging and blood and joint cultures were negative. Biopsy of the left acromion demonstrated acute and chronic osteomyelitis with areas of necrosis and granulomatous inflammation suggestive of CNO. Patient was started on high dose corticosteroids as well as methotrexate injections with marked improvement in symptoms. CONCLUSION: This case highlights that while purulent effusions are often indicative of bacterial osteomyelitis, the consideration of CNO in a patient with inflammatory bowel disease (IBD) with multifocal small bone involvement and negative blood cultures should be considered.

Refuges from fire maintain pollinator-plant interaction networks.

Fire is a major disturbance factor in many terrestrial ecosystems, leading to landscape transformation in fire-prone areas. Species in mutualistic interactions are often highly sensitive to disturbances like fire events, but the degree and complexity of their responses are unclear. We use bipartite insect-flower interaction networks across a recently burned landscape to explore how plant-pollinator interaction networks respond to a recent major fire event at the landscape level, and where fire refuges were present. We also investigate the effectiveness of these refuges at different elevations (valley to hilltop) for the conservation of displaced flower-visiting insects during fire events. Then, we explore how the degree of specialization of flower-visiting insects changes across habitats with different levels of fire impact. We did this in natural areas in the Greater Cape Floristic Region (GCFR) biodiversity hotspot, which is species rich in plants and pollinators. Bees and beetles were the most frequent pollinators in interactions, followed by wasps and flies. Highest interaction activity was in the fire refuges and least in burned areas. Interactions also tracked flower abundance, which was highest in fire refuges in the valley and lowest in burned areas. Interactions consisted mostly of specialized flower visitors, especially in refuge areas. The interaction network and species specialization were lowest in burned areas. However, species common to at least two fire classes showed no significant difference in species specialization. We conclude that flower-rich fire refuges sustain plant-pollinator interactions, especially those involving specialized species, in fire-disturbed landscape. This may be an important shelter for specialized pollinator species at the time that the burned landscape goes through regrowth and succession as part of ecosystem recovery process after a major fire event.

Tropical oribatid mites in soil toxicity testing: Optimization of test protocol and the effect of two model chemicals (cadmium and dimethoate) on Muliercula inexpectata.

The oribatid mite, Oppia nitens, has gained recognition in recent laboratory ecotoxicological tests, however, the species global distribution is limited to temperate regions and not ecologically relevant for tropical soils. The present study reports the first laboratory study aimed at assessing the ecotoxicity of contaminants with the tropical oribatid mite, Muliercula inexpectata. To develop the protocols, synchronized adult specimens of M. inexpectata were tested in a natural soil collected at the Teaching and Research Farm of the Obafemi Awolowo University, Ile-Ife, Nigeria. An optimization test was done using different soil quantities (5, 20, 40 g), durations (14, 21, 28 d) and temperature regimes (20, 22, 24, 26, and 28 °C). The results show M. inexpectata peak juvenile production at 28 d and temperatures of 24-26 °C in 20 g of soil. Test conditions were further optimized to assess the effects of cadmium and dimethoate on adult lethality and reproduction of M. inexpectata using 20 g of soil, with an exposure temperature of 26 °C for a duration of 28 d. The LC50 (survival) and EC50 (reproduction) values of cadmium for M. inexpectata were 46.55 (26.26-82.52) mg/kg and 15.61 (13.65-20.63) mg/kg, respectively. The LC50 and EC50 values of dimethoate for M. inexpectata were 7.57 (5.40-10.60) and 4.42 (0-7.16) mg/kg, respectively. Compared to other mite species, they are either more or less sensitive depending on the chemicals considered. The results of the present study demonstrate that M. inexpectata is a promising candidate for routine and ecologically-relevant ecotoxicological assessments in tropical regions.

Insect-flower interaction networks vary among endemic pollinator taxa over an elevation gradient.

Interaction networks are sensitive to elevation gradients through changes in local distribution of interacting partners. Here, we use plant-pollinator interaction network metrics to assess the effect of elevation on flowers and flower-visiting insect assemblages on a sentinel mountain used for monitoring climate change in the flower- and insect-rich Cape Floristic Region. We also use these interaction metrics to explain the effect of environmental factors on the interaction networks. We did this over four vegetation zones <1640m asl, as determined by former botanical studies. Overall, bees were the dominant flower visitors, followed by monkey beetles, and far behind were wasps and flies. The middle elevation zone (650-744 m a.s.l), which is also an ecotone between two distinct botanical zones, had the highest species richness and abundance of interacting plants and insects. Interaction frequency and size of network were also greatest in the middle zone, as were network diversity, generality, and linkage density, while lowest in the peak zone. In sum, there was distinct elevation zoning of flower-visiting insects. The greatest zonal change was between species at the middle compared with peak zone. Large-sized monkey beetles, bees and flies characterized the unique assemblage in the peak zone (1576-1640 m a.s.l.). The insect zonation tracked that of plant assemblages, with air temperature (lapse rate) being the primary driver of bee distribution, with lowest levels in the peak zone. In contrast, beetle distribution was driven mostly by flower assemblages as well as air temperature. In turn, wasp and fly interaction networks were not affected by any of the measured environmental variables. We conclude that increased elevation stress from reduced temperatures, changing abiotic weather conditions (e.g. strong winds at high elevations),and decline in flowering plant composition causes breakdown of interaction networks involving bees and beetles but not that of flies and wasps.