Design, synthesis and characterization of ethyl 3-benzoyl-7-morpholinoindolizine-1-carboxylate as anti-tubercular agents: In silico screening for possible target identification.

A thorough search for the development of innovative drugs to treat tuberculosis, especially considering the urgent need to address developing drug resistance, we report here a synthetic series of ethyl 3-benzoyl-7-morpholinoindolizine-1-carboxylate analogues (5a-o) as potent anti-tubercular agents. These morpholino-indolizines were synthesized by reacting 4-morpholino pyridinium salts, with various electron-deficient acetylenes to afford the ethyl 3-benzoyl-7-morpholinoindolizine-1-carboxylate analogues (5a-o). All synthesized intermediate and final compounds are characterized by spectroscopic methods such as 1H NMR, 13C NMR and HRMS and further examined for their anti-tubercular activity against the M. tuberculosis H37Rv strain (ATCC 27294-American type cell culture). All the compounds screened for anti-tubercular activity in the range of 6.25-50 µM against the H37Rv strain of Mycobacterium tuberculosis. Compound 5g showed prominent activity with MIC99 2.55 µg/mL whereas compounds 5d and 5j showed activity with MIC99 18.91 µg/mL and 25.07 µg/mL, respectively. In silico analysis of these compounds revealed drug-likeness. Additionally, the molecular target identification for Malate synthase (PDB 5CBB) is attained by computational approach. The compound 5g with a MIC99 value of 2.55 µg/mL against M. tuberculosis H37Rv emerged as the most promising anti-TB drug and in silico investigations suggest Malate synthase (5CBB) might be the compound's possible target.

A Narrative Review on 3D Visualization Techniques in Neurosurgical Education, Simulation and Planning.

INTRODUCTION: High-fidelity visualization of anatomical organs is crucial for neurosurgical education, simulation, and planning. This becomes much more important for minimally invasive neurosurgical procedures. Realistic anatomical visualization can allow resident surgeons to learn visual cues and orient themselves with the complex 3D anatomy. Achieving full fidelity in 3D medical visualization is an active area of research, however, the prior reviews focus on the application area and lack the underlying technical principles. Accordingly, the present study attempts to bridge this gap by providing a narrative review of the techniques used for 3D visualization. MATERIALS AND METHODS: We conducted a literature review on 3D medical visualization technology from 2018 to 2023 using the PubMed and Google Scholar search engines. The cross-referenced manuscripts were extensively studied to find literature that discusses technology relevant to 3D medical visualization. We also compiled and ran software applications that were accessible to us in order to better understand them. RESULTS: We present the underlying fundamental technology used in 3D medical visualization in the context of neurosurgical education, simulation, and planning. Further, we discuss and categorize a few important applications based on the 3D visualization techniques they use. CONCLUSIONS: The visualization of virtual human organs has not yet achieved a level of realism close to reality. This gap is largely due to the interdisciplinary nature of this research, population diversity, and validation complexities. With the advancements in computational resources and automation of 3D visualization pipelines, next-gen applications may offer enhanced medical 3D visualization fidelity.

Exploring the intersection of tuberous sclerosis and precocious puberty unveiled by hematocolpos.

We present the case of a 6-year-old girl who initially presented with acute pelvic pain, ultimately diagnosed with imperforate hymen leading to hematocolpos. Further investigation revealed additional clinical features including academic struggles, mood swings, and cutaneous findings, prompting consideration of a neurocutaneous syndrome. Magnetic Resonance Imaging (MRI) revealed features consistent with tuberous sclerosis complex (TSC), including radial migration lines in the subcortical white matter and an incidental arachnoid cyst. Notably, this case exhibited a unique presentation with absence of typical TSC findings such as subependymal nodules or cortical tubers. Additionally, precocious puberty, rarely associated with TSC, was observed, suggesting a potential link between hypothalamic lesions and hormonal imbalance. This case underscores the importance of comprehensive evaluation in pediatric patients presenting with seemingly unrelated symptoms, as it may unveil underlying conditions necessitating tailored management strategies.

An innovative reconstruction of an enbloc resected composite giant chest and abdominal wall chondrosarcoma with 3D-composite mesh.

BACKGROUND: Chest wall chondrosarcomas, although common, pose unique challenges due to their aggressive nature, rarity of abdominal wall involvement, and propensity for recurrence. We highlight the critical role of meticulous surgical planning, multidisciplinary collaboration, and innovative reconstruction techniques in achieving optimal outcomes for patients with composite giant chest and abdominal wall chondrosarcoma. CASE PRESENTATION: A 38-year-old female patient presented with progressive left chest and abdominal wall swelling for two years; on evaluation had a large lobulated lytic lesion arising from the left ninth rib, scalloping eighth and tenth ribs measuring 13.34 × 8.92 × 10.71 cm (anteroposterior/transverse/craniocaudal diameter) diagnosed with chondrosarcoma grade 2. A three-dimensional (3D) composite mesh was designed based on computed tomography using virtual surgical planning and computer-assisted design and manufacturing technology. She underwent wide local excision and reconstruction of the chest and abdominal wall with 3D-composite mesh under general anesthesia. The postoperative condition was uneventful, with no recurrence at 12 months follow-up. CONCLUSION: A 3D-composite mesh facilitates patient-specific, durable, and cost-effective chest and abdominal wall reconstruction.

Antigen identification strategies and preclinical evaluation models for advancing tuberculosis vaccine development.

In its myriad devastating forms, Tuberculosis (TB) has existed for centuries, and humanity is still affected by it. Mycobacterium tuberculosis (M. tuberculosis), the causative agent of TB, was the foremost killer among infectious agents until the COVID-19 pandemic. One of the key healthcare strategies available to reduce the risk of TB is immunization with bacilli Calmette-Guerin (BCG). Although BCG has been widely used to protect against TB, reports show that BCG confers highly variable efficacy (0-80%) against adult pulmonary TB. Unwavering efforts have been made over the past 20 years to develop and evaluate new TB vaccine candidates. The failure of conventional preclinical animal models to fully recapitulate human response to TB, as also seen for the failure of MVA85A in clinical trials, signifies the need to develop better preclinical models for TB vaccine evaluation. In the present review article, we outline various approaches used to identify protective mycobacterial antigens and recent advancements in preclinical models for assessing the efficacy of candidate TB vaccines.

4-(Benzyloxy)phenol-induced p53 exhibits antimycobacterial response triggering phagosome-lysosome fusion through ROS-dependent intracellular Ca2+ pathway in THP-1 cells.

Drug-resistant tuberculosis (TB) outbreak has emerged as a global public health crisis. Therefore, new and innovative therapeutic options like host-directed therapies (HDTs) through novel modulators are urgently required to overcome the challenges associated with TB. In the present study, we have investigated the anti-mycobacterial effect of 4-(Benzyloxy)phenol. Cell-viability assay asserted that 50 µM of 4-(Benzyloxy)phenol was not cytotoxic to phorbol 12-myristate 13-acetate (PMA) differentiated THP-1 (dTHP-1) cells. It was observed that 4-(Benzyloxy)phenol activates p53 expression by hindering its association with KDM1A. Increased ROS, intracellular Ca2+ and phagosome-lysosome fusion, were also observed upon 4-(Benzyloxy)phenol treatment. 4-(Benzyloxy)phenol mediated killing of intracellular mycobacteria was abrogated in the presence of specific inhibitors of ROS, Ca2+ and phagosome-lysosome fusion like NAC, BAPTA-AM, and W7, respectively. We further demonstrate that 4-(Benzyloxy)phenol mediated enhanced ROS production is mediated by acetylation of p53. Blocking of p53 acetylation by Pifithrin-α (PFT- α) enhanced intracellular mycobacterial growth by blocking the mycobactericidal effect of 4-(Benzyloxy)phenol. Altogether, the results showed that 4-(Benzyloxy)phenol executed its anti-mycobacterial effect by modulating p53-mediated ROS production to regulate phagosome-lysosome fusion through Ca2+ production.

Neutrophil Lymphocyte Ratio as a Predictor of Stroke Severity in Type 2 Diabetes Mellitus: A Single-Center Study.

INTRODUCTION: Type 2 diabetes mellitus (T2DM) is associated with various microvascular and macrovascular complications. Stroke, being a vascular complication, is associated with severe morbidity and mortality. Neutrophil lymphocyte ratio (NLR), a crude, inexpensive, and rather easily available modality to detect inflammation, has been utilized to find the extent of inflammation in type 2 diabetes mellitus patients. In this study, we find the effect of hemoglobin A1c (HbA1c) on NLR and the effect of NLR on stroke severity index. AIMS AND OBJECTIVES: This study aims to determine the use of the NLR in predicting stroke severity in a type 2 diabetes mellitus patient. MATERIALS AND METHODS: This study is an observational cross-sectional study. A total of 400 patients were enrolled, all of whom had type 2 diabetes mellitus, with 200 of them diagnosed with an ischemic stroke. The National Institute of Health stroke scale (NIHSS) was used to standardize stroke severity and NLR was calculated from differential counts. RESULTS: The mean NLR for patients with type 2 diabetes mellitus was 3.87 ± 0.76 (mean ± SD), while for those with type 2 diabetes mellitus and stroke, it was 7.89 ± 1.29 (mean ± SD), with a statistically significant p-value < 0.001. Additionally, for every 1 unit increase in HbA1c, the NLR increased by 0.38 in type 2 diabetes mellitus patients and 0.86 in type 2 diabetes mellitus patients with stroke. Furthermore, each 1-unit increase in NLR corresponded to a rise of 0.80 in the stroke severity index. CONCLUSION: The study shows a significant correlation between NLR in type 2 diabetes mellitus patients and stroke in type 2 diabetes mellitus patients. Also, it shows the significance of NLR in predicting stroke severity.

Polyphosphate kinase-1 regulates bacterial and host metabolic pathways involved in pathogenesis of Mycobacterium tuberculosis.

Inorganic polyphosphate (polyP) is primarily synthesized by Polyphosphate Kinase-1 (PPK-1) and regulates numerous cellular processes, including energy metabolism, stress adaptation, drug tolerance, and microbial pathogenesis. Here, we report that polyP interacts with acyl CoA carboxylases, enzymes involved in lipid biosynthesis in Mycobacterium tuberculosis. We show that deletion of ppk-1 in M. tuberculosis results in transcriptional and metabolic reprogramming. In comparison to the parental strain, the Δppk-1 mutant strain had reduced levels of virulence-associated lipids such as PDIMs and TDM. We also observed that polyP deficiency in M. tuberculosis is associated with enhanced phagosome-lysosome fusion in infected macrophages and attenuated growth in mice. Host RNA-seq analysis revealed decreased levels of transcripts encoding for proteins involved in either type I interferon signaling or formation of foamy macrophages in the lungs of Δppk-1 mutant-infected mice relative to parental strain-infected animals. Using target-based screening and molecular docking, we have identified raloxifene hydrochloride as a broad-spectrum PPK-1 inhibitor. We show that raloxifene hydrochloride significantly enhanced the activity of isoniazid, bedaquiline, and pretomanid against M. tuberculosis in macrophages. Additionally, raloxifene inhibited the growth of M. tuberculosis in mice. This is an in-depth study that provides mechanistic insights into the regulation of mycobacterial pathogenesis by polyP deficiency.

Use of the Aurolab Aqueous Drainage Implant as a buckling element in pediatric retinal detachment with a preexisting glaucoma drainage device.

Purpose: To describe a novel technique for repair of rhegmatogenous retinal detachment in an eye with a previous non-valved glaucoma drainage device, the Aurolab Aqueous Drainage Implant (AADI). Observations: A 5-year-old child with bilateral primary congenital glaucoma presented with an inferior retinal detachment (RD) in the left eye. The left eye had a history of multiple surgical interventions including combined trabeculotomy and trabeculectomy done twice, AADI implantation and subsequently phacoaspiration with IOL implantation, 18 months prior to presentation. The left eye retinal detachment was managed by scleral buckling technique using the plate of the AADI as a buckling element without its explantation. Conclusions: AND IMPORTANCE: Management of retinal detachment in eyes with a pre-existing glaucoma drainage device (GDD) is uniquely challenging. Explantation of the GDD would likely result in intractable glaucoma post-operatively, requiring another surgery. Use of the trimmed plate of the GDD itself as the buckling element helped in settling the RD and preserving intraocular pressure control.

Identification and optimization of pyridine carboxamide-based scaffold as a drug lead for Mycobacterium tuberculosis.

New drugs with novel mechanisms of action are urgently needed to tackle the issue of drug-resistant tuberculosis. Here, we have performed phenotypic screening using the Pathogen Box library obtained from the Medicines for Malaria Venture against Mycobacterium tuberculosis in vitro. We have identified a pyridine carboxamide derivative, MMV687254, as a promising hit. This molecule is specifically active against M. tuberculosis and Mycobacterium bovis Bacillus Calmette-Guérin (M. bovis BCG) but inactive against Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Escherichia coli pathogens. We demonstrate that MMV687254 inhibits M. tuberculosis growth in liquid cultures in a bacteriostatic manner. Surprisingly, MMV687254 was as active as isoniazid in macrophages and inhibited M. tuberculosis growth in a bactericidal manner. Mechanistic studies revealed that MMV687254 is a prodrug and that its anti-mycobacterial activity requires AmiC-dependent hydrolysis. We further demonstrate that MMV687254 inhibits M. tuberculosis growth in macrophages by inducing autophagy. In the present study, we have also carried out a detailed structure-activity relationship study and identified a promising novel lead candidate. The identified novel series of compounds also showed activity against drug-resistant M. bovis BCG and M. tuberculosis clinical strains. Finally, we demonstrate that in contrast to MMV687254, the lead molecule was able to inhibit M. tuberculosis growth in a chronic mouse model of infection. Taken together, we have identified a novel lead molecule with a dual mechanism of action that can be further optimized to design more potent anti-tubercular agents.

Personalized 3D Printed Eye Gear for Microscopic Surgeons Amidst and beyond COVID-19.

The COVID-19 pandemic underscored the need for enhanced protective measures for healthcare workers, particularly surgeons, who face a heightened risk of exposure to infectious aerosols. However, conventional eye protection equipment such as face shields, goggles, or glasses often leads to ergonomic discomfort and a reduced field of view (FOV), impeding surgeons' ability to perform microsurgical procedures with precision and ease. To address these limitations, this study aimed to develop personalized 3D-printed eye gear for microscopic surgeons based on facial anthropometry data. 3D scanning was employed to obtain facial data from ten neurosurgery residents. Utilizing computer-aided designing, eye gears tailored to the unique facial features of each participant were developed. Finite element analysis-based contact simulation was used to assess the pressure exerted by the eye gear. Multi-material 3D printing was employed to fabricate the personalized eye gear. Participants, while donning the eye gear, engaged in simulation-based micro suturing tasks at various magnifications of the operating microscope, and marked the FOV range. They provided feedback scores (1-10) on the effectiveness of the eye gear through a Likert scale questionnaire (Q1-Q8). Finite element analysis demonstrated uniform strain distribution on the face, indicating that the edges of the customized eye gear fit exactly to the user's face. The average scores for the questionnaire Q1 to Q8 ranged from 6.8 to 8.5, with an overall mean score of 7.6. This indicates that the developed eye gear was simple to use and did not cause any discomfort. Additionally, the average reduction in the FOV was only 10.93% across the different operating microscope magnifications. These findings highlight eye gear's potential to alleviate discomfort and enhance precision in microscopic surgeries. Consequently, personalized 3D-printed eye gear offers a promising solution for providing surgeons with a safe environment while preserving the benefits of the operating microscope.

Scrub Typhus Presenting As Acute Febrile Illness With Splenic Infarct: A Rare Manifestation.

Scrub typhus is a mite-borne infectious disease endemic in India, Korea, China, Japan, Taiwan, Pakistan, Malaysia, Thailand, and Australia. It has a multitude of clinical manifestations ranging from mild symptoms like headache, myalgia, anorexia, fever, and rash to severe multiorgan failure. It can also lead to several complications, including pancreatitis, hepatitis, myocardial infarction, and cerebral infarction. A few cases of splenic infarction are also reported. We report a rare case of a 40-year-old female presenting with fever and left upper quadrant abdominal pain of acute onset. She is diagnosed serologically with scrub typhus using enzyme-linked immunosorbent assay (ELISA) after ruling out other infectious causes, including other tropical diseases. Abdominal computed tomography revealed splenic infarction attributed to scrub typhus after excluding other etiologies. She improved after a course of doxycycline and was advised to follow up. Hence, a splenic infarct should be suspected in a patient with scrub typhus complaining of acute left hypochondriac pain.

Soybean lectin-triggered IL-6 secretion induces autophagy to kill intracellular mycobacteria through P2RX7 dependent activation of the JAK2/STAT3/Mcl-1 pathway.

Cytokine therapy and cytokine-mediated autophagy have been used as prominent host-directed therapy (HDT) approaches to restrain M. tb growth in the host cell. In the present study, we have dissected the anti-tubercular activity of Soybean lectin (SBL) through cytokine-mediated autophagy induction in differentiated THP-1 (dTHP-1) cells. A significant increase in IL-6 expression was observed in both uninfected and mycobacteria infected dTHP-1 cells through the P2RX7 mediated pathway via PI3K/Akt/CREB-dependent signalling after SBL treatment. Inhibition of IL-6 level using IL-6 neutralizing antibody or associated signalling significantly enhanced the mycobacterial load in SBL-treated dTHP-1 cells. Further, autocrine signalling of IL-6 through its receptor-induced Mcl-1 expression activated autophagy via JAK2/STAT3 pathway, and inhibition of this pathway affected autophagy. Finally, blocking the IL-6-regulated autophagy through NSC 33994 (a JAK2 inhibitor) or S63845 (an Mcl-1 inhibitor) led to a notable increase in intracellular mycobacterial growth in SBL-treated cells. Taken together, these results indicate that SBL interacts with P2RX7 to regulate PI3K/Akt/CREB network to release IL-6 in dTHP-1 cells. The released IL-6, in turn, activates the JAK2/STAT3/Mcl-1 pathway upon interaction with IL-6Rα to modulate autophagy that ultimately controls mycobacterial growth in macrophages.

What do retina fellows-in-training think about the vitreoretinal surgical simulator: A multicenter survey.

Purpose: To profile vitreoretinal (VR) fellows-in-training from India exposed to the Eyesi surgical simulator, to identify potential barriers to voluntary use, and enumerate the most preferred tools and tasks before incorporating them into a formal skill-transfer curriculum. Methods: A questionnaire consisting of 22 questions was designed and circulated through an online portal (surveymonkey.com) to four different institutes of India having a VR surgical fellowship program and using a functional Eyesi (Haag-Streit) simulator. All fellows and trainees who were exposed to the simulator were eligible to participate, irrespective of time spent on the simulator and exposure to training steps on real patients. The responses collected were private and anonymous. Results: Of the 37 respondents, most (n = 25, 68%) considered surgical simulators to be the best training tool before operating on the human eye. A majority (n = 35, 94.5%) of participants spent <3 h per week on the simulator, which, most (n = 30, 81%) felt was not enough time. The main reasons for this underutilization were work-hour limitations (54.8%), lack of a structured training program (19.3%), or a dedicated supervisor (16.1%). Again, the majority (n = 33, 89%) of participants responded that VR surgical skills acquired during simulator training were transferrable to the operating room, which was reflected by their response (n = 31, 83.7%) that simulator-based training should be made mandatory before operating room exposure. Conclusion: This study gives an insight into the overall practice patterns and preferences in simulation training of surgical VR fellows-in-training across India. It indicates that the simulator is extremely helpful to fellows and if adopted, VR surgical simulators with organized, directed, and supervised sessions will considerably improve the surgical training experience.

Ac-93253 inhibits intracellular growth of mycobacteria in human macrophages by inducing apoptosis in mitochondrial-dependent manner.

Recent studies suggest that apoptosis in macrophages plays a significant role in host defence against intracellular pathogens like viruses, fungi, protozoan, and bacteria, including Mycobacterium tuberculosis (M. tb). It is still unclear if micromolecules inducing apoptosis could be an attractive approach to combat the intracellular burden of M. tb. Hence, the present study has investigated the anti-mycobacterial effect of apoptosis mediated through phenotypic screening of micromolecules. Through MTT and trypan blue exclusion assay, 0.5 µM of Ac-93253 was found to be non-cytotoxic even after 72 h of treatment in phorbol 12-myristate 13-acetate (PMA) differentiated THP-1 (dTHP-1) cells. Significant regulation in the expression of various pro-apoptotic genes like Bcl-2, Bax, and Bad and the cleaved caspase 3 was observed upon treatment with a non-cytotoxic dose of Ac-93253. Ac-93253 treatment also leads to DNA fragmentation and increased phosphatidylserine accumulation in the plasma membrane's outer leaflet. Further, Ac-93253 also effectively reduced the growth of mycobacteria in infected macrophages, Z-VAD-FMK a broad-range apoptosis inhibitor significantly brought back the mycobacterial growth in Ac-93253 treated macrophages. These findings suggest apoptosis may be the probable effector response through which Ac-93253 manifests its anti-mycobacterial property.

Displacement of Submacular Hemorrhage With Vitrectomy Combined With Subretinal Balanced Salt Solution and Air.

PURPOSE: To describe a technique of displacement of submacular hemorrhage (SMH) using subretinal injection of balanced salt solution and filtered air. METHODS: Patients presenting within 2 weeks of massive SMH (>4 disk diameter) were prospectively included. All patients underwent 25-gauge pars plana vitrectomy, posterior vitreous detachment, injection of subretinal balanced salt solution and filtered air followed by partial fluid air exchange, 20% sulfur hexafluoride tamponade, and heads-up positioning postoperatively. Degree of displacement of SMH was assessed at 1 month and change in best-corrected visual acuity was assessed at 3 months. RESULTS: Ten patients with massive SMH who underwent the aforementioned procedure were included. Complete displacement of bleed from the macula was achieved in nine (90%) of 10 eyes at 1 month. There was significant improvement in best-corrected visual acuity from baseline at 1 month ( P = 0.015) and 3 months ( P = 0.043). CONCLUSION: Pars plana vitrectomy with injection of subretinal balanced salt solution and filtered air was well-tolerated and efficacious in displacing large and thick SMH in patients operated within 2 weeks of onset of symptoms.