Aggregated neutrophil extracellular traps occlude Meibomian glands during ocular surface inflammation.

PURPOSE: Obstructive Meibomian gland dysfunction (MGD) is one of the leading causes of evaporative dry eye disease. Meibomian glands at the eyelid secrete lipids that prevent evaporation of the aqueous tear film. The pathogenesis of obstructive MGD is incompletely understood to date. Herein, we aim to investigate the pathogenesis of obstructive MGD using murine and human samples with various forms of ocular surface inflammation. METHOD: The presence of Neutrophil extracellular Traps (NETs) was detected with immunofluorescence analysis of ocular surface discharge and biopsy samples from patients with blepharitis. Tear fluid from patients with MGD and blepharitis were evaluated for the presence of inflammatory mediators using bead based immunoassay. Murine model of allergic eye disease (AED) was performed to investigate the role of NETs in MG occlusion. RESULTS: we show that the ocular discharge from patients with blepharitis contains aggregated neutrophil extracellular traps (aggNETs). Furthermore, the ducts of human Meibomian glands affected by blepharitis were largely congested by aggNETs. Tear fluid from patients with MGD showed elevated neutrophil chemoattractants (C5a, IL6, IL8 and IL18). C5a and IL8 correlated with the degree of deficiency of tear fluid. In the murine model of allergic eye disease (AED), aggNETs accumulated in the MG leading to occlusion of their ducts and the retrograde pent-up of the fluid followed by acinar atrophy. Constraining aggNET formation by genetic or pharmacological inhibition of peptidyl arginine deiminase type 4 (PADI4) effectively reduced MG damage. CONCLUSION: We conclude that aggNETs occlude MG causing MGD after ocular surface inflammation.

Unusual case of isolated major aortopulmonary collateral artery perfusing entire functional left lower lobe of the lung.

Pulmonary sequestrations are nonfunctioning pulmonary parenchyma supplied by a systemic artery. We describe an 18-month-old baby girl with no significant past medical history who was found to have a large isolated collateral from the aorta entirely perfusing the functional left lower lobe. Cardiac catheterization confirmed the major aortopulmonary collateral artery supplying the left lower lobe with normal drainage into the left atrium. At a multidisciplinary meeting, decision was made to preserve the functional lobe. During surgery, the collateral artery was dissected and its course within the pleural space appreciated, the main left pulmonary artery was isolated. Intraoperatively surgeons anastomosed an 8 mm ringed conduit from the left pulmonary artery, anteriorly to the proximal aspect of the collateral. The patient tolerated the surgery well. Postoperative echocardiogram showed pulmonary flow to the lower lobe, and she was discharged home on postoperative day 2 without complications.

Cardiac Dysfunction and Shock in Pediatric Patients With COVID-19.

Coronavirus disease-2019 (COVID-19) has been reported to cause significant morbidity in adults, with reportedly a lesser impact on children. Cardiac dysfunction has only been described in adults thus far. We describe 3 cases of previously healthy children presenting with shock and COVID-19-related cardiac inflammation. (Level of Difficulty: Intermediate.).

Observation of Self-Bending and Focused Ultrasound Beams in the Megahertz Range.

Self-bending (or self-accelerating) and nondiffracting acoustic beams, such as Airy beams, have the potential to focus around obstacles that are directly in the beam path. Here, we demonstrate the self-bending and focusing properties of Airy beams in the ultrasound domain using finite difference time-domain simulations at 5.2 MHz. The phase profiles of self-bending Airy beams are determined from the Airy function. This beam is then transmitted experimentally using a linear array transducer connected to a 128 channel Vantage Verasonics operating at 5.2 MHz. The performance of self-bending beams is compared to conventional focused ultrasound beams in the presence of a strong scattering obstacle (steel rod). The ability of self-bending Airy beams to bypass obstacles is characterized in terms of their relative energy retention at peak intensity, that was found experimentally to be 50.5% for traditional focused beams whereas 71.5% for Airy beams, proving that self-bending beams performed better than conventional beams in terms of relative energy retention with no significant change in the focal profiles. However, it is observed that, in absolute terms, self-bending beams focus less energy than traditional focused beams.

The Median Mandibular Symphysis Tessier 30 Cleft:: 3D Imaging Analysis and Review of the Literature.

Atypical craniofacial clefts of the upper facial region have been well documented; however, the mandibular clefts remain rare and reported as isolated case reports. We report a case of a median mandibular cleft within the context of a Tessier 0-14 axis that we have followed over a 5-year period without surgical/orthodontic intervention. The mandibular symphysis cleft remained open without evidence of the fusion, in contrast to ossification of the metopic dysraphism. Within this context, we present a review of the median mandibular cleft cases from 1819 to 2015.

Lamin Cleavage: A Reliable Marker for Studying Staurosporine-Induced Apoptosis in Corneal Tissue.

Purpose: The aims of this study were to identify a robust apoptosis marker suitable for both quantification and back-to-back analyses of programmed cell death and to define specific upstream targets for apoptosis in corneal cells. Methods: Apoptotic cleavage of initiator caspases and their downstream targets such as lamins and poly-ADP ribose polymerase was investigated in human corneal endothelial cells (HCEC-12), keratocytes (HCK), epithelial cells (HCEp), and full-thickness corneas using Western blotting and confocal microscopy following apoptosis induction with staurosporine. We specifically focused on nuclear lamins, which have important structural and regulatory functions in the cell nucleus. Results: The cleavage of lamin A in HCEC-12 was significantly increased following apoptotic induction compared with HCK. More importantly, lamin A cleavage was detected in a dose-dependent manner in full-thickness corneal tissue by both Western blot analysis and fluorescence microscopy. Our study also demonstrates that HCEp show approximately three-fold increase in caspase 6 cleavage compared with endothelial cells or keratocytes. The presence of cleaved caspase 9 was lower in endothelial cells compared with epithelial cells and keratocytes. Conclusions: We successfully established lamin A cleavage as a quantifiable marker of apoptosis in both corneal cells and tissue. Quantification of lamin A cleavage by Western blotting followed by a back-to-back analysis with fluorescence microscopy was studied for the first time in the experimental (donor) corneal tissue. Screening of downstream apoptosis proteins and establishing cell type-specific protocols allowed us to identify possible targets (caspases, Apaf-1, etc.) for protective therapeutic approaches.

Genome sequence of a clinical isolate of dermatophyte, Trichophyton rubrum from India.

Trichophyton rubrum is one of the major causative agents of dermatophytosis in humans worldwide. We report the draft genome sequence of T. rubrum var. raubitschekii from Delhi, India, isolated from a patient presenting symptoms of onychomycosis. The total estimated genome size of the clinical isolate is 25.2 MB containing 8265 predicted protein-coding sequences, 91 tRNA and 15 rRNA genes. Sequence analysis of the secreted subtilases, one of the major virulence factors in dermatophytes, clusters them into three subfamilies with distinct sequence features. The genome sequence is a step in understanding diversity of dermatophytes worldwide and will aid in identification of virulence factors and dissecting mechanisms of pathogenesis among them.

Preimplantation and postimplantation therapy for the treatment of reproductive failure.

Treatment of patients with recurrent pregnancy losses and recurrent implantation failure can be instituted only when the underlying etiology is determined. Embryo-secreted preimplantation factor (PIF) is essential for implantation and adequate trophoblastic invasion. Deficiency of PIF affects the outcome of the pregnancy leading to recurrent pregnancy losses. Synthetic PIF modulates the outcome of the pregnancy decreasing the incidence of recurrent implantation failure and recurrent pregnancy losses. In this article a thorough search is done regarding the data published for diagnoses of reproductive failure and its treatment. The effect of immunoglobulin (Ig), intralipid, heparin, aspirin, progesterone, estrogen, and granulocyte colony stimulating factor (G-CSF) is taken into consideration. Heparin, aspirin, and progesterone have successfully shown to decrease the incidence of recurrent pregnancy loses; whereas G-CSF, intralipids, estrogen, and Igs have shown success in the treatment of the recurrent implantation failure and recurrent pregnancy failure. The pregnancies treated with Igs and intralipids showed equal outcome when evaluated and compared. The place of intralipid in reducing natural killer (NK) cells has been discussed.

Isolation, identification and optimization of a new extracellular lipase producing strain of Rhizopus sp.

A lipolytic mesophilic fungus which produces lipase extracellularly was isolated from soil. Based on ITS1-5.8S-ITS4 region sequences of ribosomal RNA, it was concluded that the isolate JK-1 belongs to genus Rhizopus and clades with Rhizopus oryzae. The present paper reports the screening, isolation, identification, and optimization of fermentation conditions for the production of lipase (EC 3.1.1.3). Culture conditions were optimized, and the highest lipase production was observed in basal medium with corn steep liquor as nitrogen source and glucose as carbon source. Maximum lipase production was observed at 72 h, which is about 870 U/ml. Optimization of fermentation conditions resulted in 16-fold enhancement in enzyme production.

Novel point mutations in sulfadoxine resistance genes of Plasmodium falciparum from India.

Point mutations in the dhfr and dhps genes of Plasmodium falciparum are associated with pyrimethamine and sulfadoxine resistance respectively. In this study we have analyzed these genes from Bikaner (situated in North-West region of India), where both uncomplicated and severe manifestations of P. falciparum malaria are seen. A majority of isolates showed double mutant allele for DHFR. In contrast, the only reported mutation present in DHPS was A437G in few isolates. In addition, three novel non-synonymous mutations were observed in the PfDHPS from this region viz., S587F, N666K and C668W. The mutations at the 666 and 668 codon seem to form a bend in the big loop region of the DHPS enzyme and may affect the binding of the drug to the enzyme. Molecular docking of sulfadoxine to this mutated structure indicates reduction in its binding affinity to this enzyme.