Noncanonical role for Ku70/80 in the prevention of allergic airway inflammation via maintenance of airway epithelial cell organelle homeostasis.

Airway epithelial homeostasis is under constant threat due to continuous exposure to the external environment, and abnormally robust sensitivity to external stimuli is critical to the development of airway diseases, including asthma. Ku is a key nonhomologous end-joining DNA repair protein with diverse cellular functions such as VDJ recombination and telomere length maintenance. Here, we show a novel function of Ku in alleviating features of allergic airway inflammation via the regulation of mitochondrial and endoplasmic reticulum (ER) stress. We first determined that airway epithelial cells derived from both asthmatic lungs and murine asthma models demonstrate increased expression of 8-hydroxy-deoxyguanosine (8-OHdG), a marker of oxidative DNA damage. Ku protein expression was dramatically reduced in the bronchial epithelium of patients with asthma as well as in human bronchial epithelial cells exposed to oxidative stress. Knockdown of Ku70 or Ku80 in naïve mice elicited mitochondrial collapse or ER stress, leading to bronchial epithelial cell apoptosis and spontaneous development of asthma-like features, including airway hyperresponsiveness, airway inflammation, and subepithelial fibrosis. These findings demonstrate an essential noncanonical role for Ku proteins in asthma pathogenesis, likely via maintenance of organelle homeostasis. This novel function of Ku proteins may also be important in other disease processes associated with organelle stress.

Iatrogenic Bilateral Simultaneous Pneumothorax: Call for Vigilance.

Iatrogenic pneumothorax refers to the pneumothorax generated after diagnostic or therapeutic procedure. We report the case of a 40-year-old male who had bilateral simultaneous iatrogenic pneumothorax with pneumomediastinum leading to cardiac arrest situation, due to wrong placement of nebulization kit in spontaneously breathing intubated patient. We report this case for its rarity, due to the critical importance of this cause as a etiology of bilateral simultaneous iatrogenic pneumothorax leading to cardiac arrest situation, and need to understand the importance of continuous training and stress of emergency environment.

Parabromophenacyl bromide inhibits subepithelial fibrosis by reducing TGF-ß1 in a chronic mouse model of allergic asthma.

BACKGROUND: Our previous study showed that parabromophenacyl bromide (PBPB) inhibits the features of allergic airway inflammation and airway hyperresponsiveness (AHR). However, its effect on airway remodeling, e.g. subepithelial fibrosis in a chronic allergic asthma model, was not investigated. We examined this issue in this study. METHODS: PBPB was administered to mice with an induced chronic asthmatic condition. AHR was estimated at the end of the experiment, followed by euthanasia. Lung sections were stained with hematoxylin and eosin, periodic acid-Schiff and Masson's trichrome to determine airway inflammation, goblet cell metaplasia and subepithelial fibrosis, respectively. Transforming growth factor-ß1 (TGF-ß1) was estimated in lung homogenates. To determine the effect of PBPB on smooth-muscle hyperplasia, immunohistochemistry against α-smooth-muscle actin was performed on the lung sections. RESULTS: Chronic ovalbumin challenges in a mouse model of allergic asthma caused significant subepithelial fibrosis and elevated TGF-ß1, along with significant AHR. PBPB attenuated subepithelial fibrosis with a reduction of lung TGF-ß1, airway inflammation and AHR without affecting goblet cell metaplasia. It also attenuated smooth-muscle hyperplasia with a reduction in the expression of α-smooth-muscle actin in the lungs. CONCLUSION: Our findings indicate that PBPB attenuates some crucial features of airway remodeling such as subepithelial fibrosis and smooth-muscle hyperplasia. These data suggest that PBPB could therefore be a therapeutic drug for chronic asthma.

Dental pain and self-care: a cross-sectional study of people with low socio-economic status residing in rural India.

CONTEXT: Self-care is one of the ways in which people without access to professional care can actively engage in managing their oral health problems. AIM: To find out the prevalence of dental pain and the type of self-care remedies utilised for pain relief by people with low socio-economic status residing in Kollipara mandal, Guntur district, Andhra Pradesh. SETTINGS AND DESIGN: This was a cross-sectional population-based study. METHODS AND MATERIAL: A multistage simple random-sampling technique was adopted to obtain an appropriate sample. A questionnaire, which consisted of sections on socio-economic and demographic variables, dental pain, pain characteristics and self-care remedies utilised to combat dental pain, was used to collect data. Statistical analysis used the chi-square test and descriptive statistics. RESULTS: The survey covered a total population of 630 individuals. The mean age of the population was 32.8±16.7 years. Among the total study subjects, 44.1% were male and 55.9% were female. The prevalence of dental pain reported during the preceding 6 months was 28.3%. Subjects who experienced dental pain reported the home remedies and self-care methods that they utilised to get relief from the pain. They more frequently reported using over-the-counter medication (49.6%) for pain relief. CONCLUSIONS: This study provides an insight into the type and usage of self-care in relief of dental pain. Pain sufferers used a variety of self-care methods to deal with their problems.

Dental manpower in India: changing trends since 1920.

AIM: To analyse the changing trends in dental manpower production of India since 1920 and its development to date. METHODS AND MATERIAL: The databases consulted were those provided by the Central Bureau of Health Intelligence, Dental Council of India, and Ministry of Health and Family Welfare. STATISTICAL ANALYSIS USED: Descriptive statistics. RESULTS: In India, dental education was formally established in 1920 when the first dental college was started. Current data revealed that there are 301 colleges nationwide granting degrees in dentistry, with a total of 25,270 student positions offering annually. Both the distribution of dental colleges and of dentists varies among the regions of the country with the greatest concentration in major urban areas, resulting in limited coverage in rural regions. CONCLUSIONS: The current scenario indicates that there is lack of systematic planning in the allocation and development of dental colleges in India.

Macroalgal deep genomics illuminate multiple paths to aquatic, photosynthetic multicellularity.

Macroalgae are multicellular, aquatic autotrophs that play vital roles in global climate maintenance and have diverse applications in biotechnology and eco-engineering, which are directly linked to their multicellularity phenotypes. However, their genomic diversity and the evolutionary mechanisms underlying multicellularity in these organisms remain uncharacterized. In this study, we sequenced 110 macroalgal genomes from diverse climates and phyla, and identified key genomic features that distinguish them from their microalgal relatives. Genes for cell adhesion, extracellular matrix formation, cell polarity, transport, and cell differentiation distinguish macroalgae from microalgae across all three major phyla, constituting conserved and unique gene sets supporting multicellular processes. Adhesome genes show phylum- and climate-specific expansions that may facilitate niche adaptation. Collectively, our study reveals genetic determinants of convergent and divergent evolutionary trajectories that have shaped morphological diversity in macroalgae and provides genome-wide frameworks to understand photosynthetic multicellular evolution in aquatic environments.

Cellular Distribution of Secreted Phospholipase A2 in Lungs of IPF Patients and Its Inhibition in Bleomycin-Induced Pulmonary Fibrosis in Mice.

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease with a very poor prognosis as it has a 2.5 to 5 years mean survival after proper diagnosis. Even nintedanib and pirfenidone cannot halt the progression, though they slow the progression of IPF. Hence, there is a need to understand the novel pathophysiology. Phospholipase A2 (PLA2) could be the ideal candidate to study in IPF, as they have a role in both inflammation and fibrosis. In the present study, we have shown the expression profile of various secretory Phospholipase A2 (PLA2) isoforms by analyzing publicly available transcriptome data of single cells from the lungs of healthy individuals and IPF patients. Among 11 members of sPLA2, PLA2G2A is found to be increased in the fibroblasts and mesothelial cells while PLA2G5 is found to be increased in the fibroblasts of IPF patients. We identified a subset of fibroblasts expressing high PLA2G2A with moderate expression of PLA2G5 and which are specific to IPF only; we named it as PLA2G2A+ IPF fibroblast. Pathway analysis revealed that these PLA2G2A+ IPF fibroblast have upregulation of both inflammatory and fibrosis-related pathways like the TGF-ß signaling pathway, IL-17 signaling, the arachidonic acid metabolism pathway and ECM-receptor interaction. In addition to this, we found elevated levels of sPLA2-IIA in plasma samples of IPF patients in our cohort. PLA2G3, PLA2G10 and PLA2G12B are found in to be increased in certain epithelial cells of IPF patients. Thus, these findings indicate that these five isoforms have a disease-dominant role along with innate immune roles as these isoforms are found predominantly in structural cells of IPF patients. Further, we have targeted sPLA2 in mice model of bleomycin-induced lung fibrosis by pBPB, a known sPLA2 inhibitor. pBPB treatment attenuated lung fibrosis induced by bleomycin along with a reduction in TGF-ß and deposition of extracellular matrix in lung. Thus, these findings indicate that these sPLA2 isoforms especially PLA2G2A may serve as a therapeutic target in lung fibrosis.

Primary clear cell carcinoma of lung presented with generalized lymphadenopathy mimicking lymphoma.

Primary Clear cell carcinoma of lung with distant metastasis is a rare tumour. Here is a case of 45 year old male presented with gradual onset dyspnoea, low grade fever and weight loss. Radiologically patient had hilar and parahilar lesion at posterior mediastinum with mild changes of chronic obstructive pulmonary disease with right supraclavicular, peripancreatic and celiac axis lymph nodes enlargement. Fine needle aspiration from lymph node followed by excision biopsy was done. For further localization Fibre optic Bronchoscopy was done which is followed by bronchial wash cytology and transbronchial needle aspiration and bronchial biopsy. Correlating all above diagnostic modalities diagnosis of metastatic clear cell adenocarcinoma of lung was made which is further supported by Immunohistochemistry.

Novel benzoxazinone derivative as potent human neutrophil elastase inhibitor: Potential implications in lung injury.

Neutrophil elastase, a powerful physiological defence tool, may serve as drug target for diverse diseases due to its bystander effect on host cells like chronic obstructive pulmonary disease (COPD). Here, we synthesised seven novel benzoxazinone derivatives and identified that these synthetic compounds are human neutrophil elastase inhibitor that was demonstrated by enzyme substrate kinetic assay. One such compound, PD05, emerged as the most potent inhibitor with lower IC50 as compared to control drug sivelestat. While this inhibition is competitive based on substrate dilution assay, PD05 showed a high binding affinity for human neutrophil elastase (Kd = 1.63 nM) with faster association and dissociation rate compared to notable elastase inhibitors like ONO 6818 and AZD9668, and its interaction with human neutrophil elastase was fully reversible.Preclinical pharmacokinetic studies were performed in vitro where protein binding was found to be 72% with a high recovery rate, aqueous solubility of 194.7 µM, low permeability along with a favourable hERG. Experiments with cell line revealed that the molecule successfully prevented elastase induced rounding and retracted cell morphology and cell cytotoxicity. In mouse model PD05 is able to reduce the alveolar collapse induced by neutrophil elastase. In summary, we demonstrate the in situ, in vitro and in vivo anti-elastase potential of the newly synthesised benzoxazinone derivative PD05 and thus this could be promising candidate for further investigation as a drug for the treatment of COPD.

Ku70 modulation alleviates murine allergic asthma features and restores mitochondrial function in lungs.

The airway epithelium is continuously exposed to a variety of pollutants and allergens, thanks to both natural and manmade environmental pollution. With numerous protective mechanisms, the airway epithelium protects the lungs. DNA repair mechanism is one such protective response and its failure could lead to the accumulation of DNA mutations. Our lab had earlier demonstrated the dysfunctional mitochondria in airway epithelium of the asthmatic mice lungs. Here, we show that Ku70 modulation by the administration of Ku70 plasmid attenuates asthma features and reduces mitochondrial dysfunction in the lungs of allergen exposed mice. Ku70 is a key DNA repair protein with diverse roles including VDJ recombination, telomere maintenance, and maintenance of cell homeostasis. Recently, we found a reduction in Ku70 expression in asthmatic airway epithelium, and this was associated with mitochondrial dysfunction in asthmatic condition. In this study, we have shown that Ku70 over-expression in asthmatic mice attenuated airway hyperresponsiveness, airway inflammation, sub-epithelial fibrosis along with reduction in TGF-ß with no effect in IL-13 levels and goblet cell metaplasia. Ku70 over-expression in asthmatic mice reduced 8-isoprostane, a marker of oxidative stress, and restored the mitochondrial function in asthmatic mice. We further found these roles of Ku70 to be independent of DNA damage as Ku70 overexpressed mice did not show any reduction in DNA tail, an index of DNA damage. Thus, our findings indicate that Ku70 can attenuate crucial features of asthma along with the restoration of mitochondrial function. This implies that Ku70 could be a therapeutic target for asthma without affecting DNA repair function.

Neuroendocrine Carcinoma of the Colon: Emergency Presentation of a Rare Disease With Poor Biology.

Neuroendocrine tumors (NETs) are rare entities. Most common among them are gastroenteropancreatic neuroendocrine tumors (GEP-NETs) and pulmonary NETs. Most of them are indolent in nature. Colonic NETs are rare among GEP-NETs and mostly present with large size and with metastasis. Emergency presentation with hematochezia is rare in colonic NETs. This case report discusses a rare emergency presentation of colonic NETs and highlights their poor biological nature.

Minimally Invasive L5 Corpectomy with Navigated Expandable Vertebral Cage: A Technical Note.

BACKGROUND: Conventional L5 corpectomy requires a large incision and an extended period of intraoperative fluoroscopy. We describe herein a new L5 corpectomy technique. METHODS: A 79-year-old woman was referred to our hospital for leg pain and lower back pain due to an L5 vertebral fracture. Her daily life had been affected by severe lower back pain and sciatica for more than 2 months. We initially performed simple decompression surgery, but this proved effective for only 10 months. RESULTS: For revision surgery, the patient underwent minimally invasive L5 corpectomy with a navigated expandable cage without fluoroscopy. The second surgery took 215 min, and estimated blood loss was 750 mL. The revision surgery proved successful, and the patient could then walk using a cane. In terms of clinical outcomes, the Oswestry Disability Index improved from 66% to 24%, and the visual analog scale score for lower back pain improved from 84 to 31 mm at the 1-year follow-up. CONCLUSIONS: Minimally invasive L5 corpectomy with a navigated expandable vertebral cage is effective for reducing cage misplacement and surgical invasiveness. With this new technique, surgeons and operating room staff can avoid the risk of adverse events due to intraoperative radiation exposure.

Protocol to generate and characterize biofouling transformants of a model marine diatom.

Diatoms are a major group of microalgae that initiate biofouling by surface colonization of human-made underwater structures; however, the involved regulatory pathways remain uncharacterized. Here, we describe a protocol for identifying and validating regulatory genes involved in the morphology shift of the model diatom species Phaeodactylum tricornutum during surface colonization. We also provide a workflow for characterizing biofouling transformants. By using this protocol, gene targets such as GPCR signaling genes could be identified and manipulated to turn off diatom biofouling. For complete information on the generation and use of this protocol, please refer to Fu et al. (2020).

Large-scale genome sequencing reveals the driving forces of viruses in microalgal evolution.

Being integral primary producers in diverse ecosystems, microalgal genomes could be mined for ecological insights, but representative genome sequences are lacking for many phyla. We cultured and sequenced 107 microalgae species from 11 different phyla indigenous to varied geographies and climates. This collection was used to resolve genomic differences between saltwater and freshwater microalgae. Freshwater species showed domain-centric ontology enrichment for nuclear and nuclear membrane functions, while saltwater species were enriched in organellar and cellular membrane functions. Further, marine species contained significantly more viral families in their genomes (p = 8e-4). Sequences from Chlorovirus, Coccolithovirus, Pandoravirus, Marseillevirus, Tupanvirus, and other viruses were found integrated into the genomes of algal from marine environments. These viral-origin sequences were found to be expressed and code for a wide variety of functions. Together, this study comprehensively defines the expanse of protein-coding and viral elements in microalgal genomes and posits a unified adaptive strategy for algal halotolerance.

Alternative glycosylation controls endoplasmic reticulum dynamics and tubular extension in mammalian cells.

The endoplasmic reticulum (ER) is a central eukaryotic organelle with a tubular network made of hairpin proteins linked by hydrolysis of guanosine triphosphate nucleotides. Among posttranslational modifications initiated at the ER level, glycosylation is the most common reaction. However, our understanding of the impact of glycosylation on the ER structure remains unclear. Here, we show that exostosin-1 (EXT1) glycosyltransferase, an enzyme involved in N-glycosylation, is a key regulator of ER morphology and dynamics. We have integrated multiomics and superresolution imaging to characterize the broad effect of EXT1 inactivation, including the ER shape-dynamics-function relationships in mammalian cells. We have observed that inactivating EXT1 induces cell enlargement and enhances metabolic switches such as protein secretion. In particular, suppressing EXT1 in mouse thymocytes causes developmental dysfunctions associated with the ER network extension. Last, our data illuminate the physical and functional aspects of the ER proteome-glycome-lipidome structure axis, with implications in biotechnology and medicine.

Amyopathic dermatomyositis with diffuse lung disease.

A 38-year-old woman presented with progressively increasing breathlessness, recurrent productive cough, and intermittent fever of 1 year duration. Examination revealed cutaneous eruptions on the dorsal aspects of the hands and on face. Histopathologic features of skin biopsy revealed acanthosis, hyperkeratosis with focal vacuolar alteration of the basal-cell layer, and perivascular inflammatory infiltrates in upper dermis. CT scan showed diffuse lung disease and pulmonary function tests showed severe restrictive lung disease. There was no muscular involvement clinically or on electromyography and magnetic resonance imaging. She was diagnosed as a case of amyopathic dermatomyositis with diffuse lung disease and managed with topical and systemic steroid and topical sunscreen with fairly good response.

A looming role of mitochondrial calcium in dictating the lung epithelial integrity and pathophysiology of lung diseases.

With the increasing appreciation of mitochondria in modulating cellular homeostasis, various disease biology researchers have started exploring the detailed role of mitochondria in multiple diseases beyond neuronal and muscular diseases. In this context, emerging shreds of evidence in lung biology indicated the meticulous role of lung epithelia in provoking a plethora of lung diseases in contrast to earlier beliefs. As lung epithelia are ceaselessly exposed to the environment, they need to have multiple protective mechanisms to maintain the integrity of lung structure and function. As ciliated airway epithelium and type 2 alveolar epithelia require intense energy for executing their key functions like ciliary beating and surfactant production, it is no surprise that defects in mitochondrial function in these cells could perturb lung homeostasis and engage in the pathophysiology of lung diseases. On one hand, intracellular calcium plays the central role in executing key functions of lung epithelia, and on the other hand maintenance of intracellular calcium needs the buffering role of mitochondria. Thus, the regulation of mitochondrial calcium in lung epithelia seems to be critical in lung homeostasis and could be decisive in the pathogenesis of various lung diseases.

GPCR Genes as Activators of Surface Colonization Pathways in a Model Marine Diatom.

Surface colonization allows diatoms, a dominant group of phytoplankton in oceans, to adapt to harsh marine environments while mediating biofoulings to human-made underwater facilities. The regulatory pathways underlying diatom surface colonization, which involves morphotype switching in some species, remain mostly unknown. Here, we describe the identification of 61 signaling genes, including G-protein-coupled receptors (GPCRs) and protein kinases, which are differentially regulated during surface colonization in the model diatom species, Phaeodactylum tricornutum. We show that the transformation of P. tricornutum with constructs expressing individual GPCR genes induces cells to adopt the surface colonization morphology. P. tricornutum cells transformed to express GPCR1A display 30% more resistance to UV light exposure than their non-biofouling wild-type counterparts, consistent with increased silicification of cell walls associated with the oval biofouling morphotype. Our results provide a mechanistic definition of morphological shifts during surface colonization and identify candidate target proteins for the screening of eco-friendly, anti-biofouling molecules.

A rare case of bilateral luxatio erecta with bilateral greater tuberosity fracture following a fall due to seizure.

Luxatio erecta is the least common form of shoulder dislocations. Bilateral luxatio erecta with concomitant greater tuberosity fracture is further rare with only couple of cases reported so far in the literature. We present an uncommon and a rare case of post epileptic seizure related bilateral luxatio erecta with greater tuberosity fracture in a young Indian male which is unique in its etiology, method of management and rarity of its type among so far reported cases.

Feasibility and Safety of Outpatient Lumbar Microscopic Discectomy in a Developing Country.

Study Design: Prospective study. Purpose: To verify the feasibility and safety of outpatient microscopic lumbar discectomy (MLD) in a developing country. Overview of Literature: Outpatient MLD is advantageous in terms of cost effectiveness and avoidance of nosocomial infections. Safety of outpatient MLD has been well established in the developed nations of North America and Europe. There is no published study of outpatient MLD from the rest of the world, especially in developing countries. Methods: Fifty-eight consecutive patients undergoing outpatient MLD with a median follow-up time of 12 months (range, 6-21 months) were included in this study. Simultaneous patient counseling was done by a surgical and anesthetic team preoperatively and pre-discharge. We collected and analyzed data pertaining to the demography, socioeconomic status, perioperative parameters, complications, and outcome assessment scores of the patients. Results: The average patient age was 37.8±9.6 years (39 males, 19 females). Unilateral discectomy was performed in 55 patients, and bilateral discectomy in three. The majority (80.3%) of the patients were classified to lower middle (III) or upper lower (IV) class on the Modified Kuppuswamy Scale. The average operative time was 41.0±8.4 minutes with an average blood loss of 42.6±14.9 mL. The average postoperative stay was 5.5±0.7 hours and the successful discharge rate was 100%. Complications noted were postoperative nausea (n=8), urinary retention (n=2), meralgia paresthetica (n=3), delayed wound healing (n=2), and recurrence (n=1). The successful outcome rates were Visual Analog Scale (VAS) score leg pain, 93.1%; VAS score back pain, 89.6%; Oswestry Disability Index score, 91.3%; return to activities of daily living, 94.8%; return to work, 79.3%; patient satisfaction rate, 82.7%; and overall success rate, 88.4%. Conclusions: Outpatient MLD can be safely performed with success, even in the setting of a developing country, if the prerequisites of appropriate patient selection, arduous adherence to outpatient surgery protocol, competent surgical/anesthetic team, and infrastructure needed for conduction of microsurgery are met.