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.

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.

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.

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.

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.

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.

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.

Potential for Heightened Sulfur-Metabolic Capacity in Coastal Subtropical Microalgae.

The activities of microalgae support nutrient cycling that helps to sustain aquatic and terrestrial ecosystems. Most microalgal species, especially those from the subtropics, are genomically uncharacterized. Here we report the isolation and genomic characterization of 22 microalgal species from subtropical coastal regions belonging to multiple clades and three from temperate areas. Halotolerant strains including Halamphora, Dunaliella, Nannochloris, and Chloroidium comprised the majority of these isolates. The subtropical-based microalgae contained arrays of methyltransferase, pyridine nucleotide-disulfide oxidoreductase, abhydrolase, cystathionine synthase, and small-molecule transporter domains present at high relative abundance. We found that genes for sulfate transport, sulfotransferase, and glutathione S-transferase activities were especially abundant in subtropical, coastal microalgal species and halophytic species in general. Our metabolomics analyses indicate lineage- and habitat-specific sets of biomolecules implicated in niche-specific biological processes. This work effectively expands the collection of available microalgal genomes by ∼50%, and the generated resources provide perspectives for studying halophyte adaptive traits.

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.

Tuberculosis of gallbladder neck: a cause of cholecystitis with cholelithiasis.

Tuberculosis of gallbladder neck is not a very common problem reported in the literature. Here, we report a case of gallbladder neck tuberculosis complicated with chronic cholecystitis with cholelithiasis in a 55-year-old woman. Diagnosis was made postoperatively on surgical biopsy.

Pulmonary nocardiosis--a rare case of pulmonary nocardiosis in an immunocompetent patient.

Nocardiosis is localised or disseminated infection caused by soil dwelling aerobic actinomycetes, which habitually enter through the respiratory tract. There has been an increase in the incidence of nocardia species infections probably due to higher degree of clinical suspicion, aggressive diagnostic examinations, increased use of immunosuppressive treatments (chemotherapy agents and immunosuppressive agents) and the appearance of AIDS. Here we are presenting an atypical case of Nocardia asteroides in a 45-years-old immunocompetent female patient presenting with history of cough with scanty expectoration, haemoptysis and fever for 4 months. Clinical and radiological diagnostic consideration were tuberculosis and malignancy. Diagnosis of nocardiosis was confirmed microbiologically and the patient responded to cotrimoxazole.

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.

Role of intraoperative Iso-C based navigation in challenging spine trauma.

BACKGROUND: Pedicle screw fixation is the most preferred method of stabilizing unstable spinal fractures. Pedicle screw placement may be difficult in presence of fractured posterior elements, deformed spine, gross instability and spinal pathology. Challenging spine-fracture fixation is defined as the presence of one or more of the following: 1) obscured topographical landmarks as in ankylosing spondylitis, 2) fractures in occipitocervical or cervicothoracic regions and 3) preexisting altered spinal alignment. We report a series of pedicle screw insertion with guidance of navigation in difficult fixation problems.. MATERIALS AND METHODS: Fourteen patients [hangman's fracture (n=3), odontoid fracture (n=4), C1C2 fracture (n=1) and spinal fracture with coexistent ankylosing spondylitis (n=6)] underwent posterior stabilization. Intraoperatively after surgical exposure, images were acquired by Iso-C 3D C-arm and transferred to navigation system. Instrumentation was performed with navigational assistance. Postoperatively, placements of pedicle screws were evaluated with radiographs and CT scan. RESULTS: Sixty-seven pedicle screws (cervical, n=33; thoracic, n=6; lumbar, n=26; sacral n=2) and 15 lateral mass screws were inserted with navigation guidance. The average time of image data acquisition by Iso-C 3D C-arm and its transfer to workstation was 4 minutes (range, 2-6 minutes). Postoperative CT scan revealed ideal placement of screws in 63 pedicles (94%), grade 1 cortical breaches (<2 mm) in 3 pedicles (4.5%) and grade 2 cortical breach (2-4 mm) in one pedicle (1.5%). There were no neurovascular complications. Deep infection was encountered in one case, which settled with debridement. CONCLUSIONS: Intraoperative Iso-C 3D C-arm based navigation is a useful adjunct while stabilizing challenging spinal trauma, rendering feasibility, accuracy and safety of pedicle screw placement even in difficult situations.