Tear film parameters and meibography in eyes of children with vernal keratoconjunctivitis.

PURPOSE: To study tear film parameters and Meibomian glands in pediatric patients with vernal keratoconjunctivitis (VKC). METHODS: Forty-four eyes of 22 patients with VKC and 44 eyes of 22 age-matched controls were studied in a hospital-based cross-sectional study. Each subject underwent a complete ophthalmological examination including slit-lamp biomicroscopy, non-invasive break-up time (NIBUT), fluorescein break-up time (FBUT), corneal fluorescein staining, and Schirmer's test. Non-contact meibography was performed using a Sirius topographer. RESULTS: All patients had VKC grades between 1 and 3 (Bonini scale). The mean NIBUT for cases was 8.83 ± 4.02 s and in controls, it was 15 ± 2.8 s (P value < 0.001). The mean FBUT for cases was 9.41 ± 3.45 s and in controls, it was 16.7 ± 2.5 s (P value < 0.001). Schirmer's values were 32 ± 3.1 mm in cases and 21.4 ± 4.1 mm in controls, which were significantly higher (P value < 0.001). Also, 22.72% of the cases had corneal fluorescein staining in the form of superficial punctuate keratitis, whereas none of the control groups showed corneal staining. The examination of Meibomian glands using non-contact meibography showed a mean Meibomian gland dropout in the upper lid to be 29.7 ± 14.4% in VKC cases and 10.5 ± 3.2% in controls (P value < 0.001). Meibography of the lower lid showed 25.9 ± 10.2% dropout in cases and 20.5 ± 6.1% in controls (P value = 0.004). CONCLUSION: Tear film parameters were significantly deranged in children with VKC in the active stage as compared to age-matched controls. Children with VKC also had a higher Meibomian gland dropout.

Agreement between Phenotypically Detected Linezolid Resistance and Mutations in rrl and rplC Genes of Mycobacterium tuberculosis Isolates Using Nanopore Sequencing.

BACKGROUND: Phenotypic drug susceptibility testing (DST) is considered the gold standard for detecting linezolid (LZD) resistance in Mycobacterium tuberculosis (MTB), but it is time-consuming. Nanopore sequencing offers a potentially faster alternative approach. This study evaluated the agreement between phenotypically detected LZD resistance and mutations in the rrl and rplC genes of MTB isolates using nanopore sequencing. METHODS: Consecutive drug-resistant MTB isolates from pulmonary samples collected in 2021 underwent liquid culture (LC) DST for LZD. All resistant isolates and an equal number of susceptible isolates were subjected to targeted sequencing of the rrl and rplC genes using nanopore technology. RESULTS: Sequencing identified a C154R mutation in the rplC gene in only one LZD-resistant isolate. No mutations were detected in the rrl gene. The agreement between sequencing and LC-DST for detecting LZD resistance was poor (Cohen's kappa: 0.03571, 95% confidence interval [CI]: -0.034-0.105). Additionally, no significant association was found between LZD resistance and clinical or microbiological outcomes at 6-month follow-up. CONCLUSION: This study revealed a considerable discrepancy between phenotypic and genotypic detection of LZD resistance in MTB. Further research is needed to better understand the genetic mechanisms underlying LZD resistance and to develop reliable molecular diagnostics for rapid resistance detection.

Efficacy of TrueNat cartridge extracted DNA for detecting drug resistant tuberculosis by line probe assay.

BACKGROUND: Currently for diagnosing Mycobacterium tuberculosis and its drug resistance, two sputum samples are required. One of them is subjected to TrueNat™ and if positive the other sample is subjected to line probe assay (LPA). This study was done to evaluate whether TrueNat extracted DNA can be directly used for performing LPA in a diagnostic laboratory setting to decrease patient turn-around time. METHODS: Total 45 smear positive sputum samples were subjected to TrueNat™ MTB detection and first and second line (FL and SL) LPA testing in parallel. DNA extracted by Trueprep® Cartridge was also tested by LPA and results were compared. Further, TrueNat extracted DNA from 20 samples was divided into 6 aliquots each, two of which were stored at 4 °C, 37 °C and 55 °C (under humidification) each. One aliquot from stored DNA at each temperature was used for FL & SL LPA on day three and the other on day eight. The blots thus obtained were compared with those of conventional LPA at day 1. RESULTS: For FL-LPA, TrueNat extracted DNA gave valid results for all 45 (100%) samples but conventionally extracted DNA could give results for 44 (97.8%) samples. Likewise, for SL-LPA, valid results were obtained for 40 (88.9%) and 35 (77.8%) samples respectively using TrueNat extracted DNA and conventionally extracted DNA respectively. All samples with invalid LPA results had Ct values ≥ 28 by TrueNat PCR. LPA results were obtained for all the 20 samples using stored DNA at all temperatures and duration. CONCLUSIONS: TrueNat extracted DNA can be used for performing LPA under field conditions for selected samples.

Normative Values of Cerebral Blood Flow Velocities in Very Low Birth Weight Neonates During First 28 Days of Life.

OBJECTIVE: To determine the normative values of cerebral blood flow (CBF) velocities in very low birth weight (VLBW) neonates during the first 28 days of life. METHOD: In this prospective observational study, doppler assessment of CBF velocities was performed from the anterior cerebral artery (ACA), middle cerebral artery (MCA) and basilar artery (BA) at 2-8 hours, 24 hours, day 3, 7, 14 and 28 of life. Neonates with gross congenital malformations, those requiring extensive resuscitation at birth, mechanical ventilation with mean airway pressure >12 mbar, requiring inotropes, or those who developed intraventricular hemorrhage (grade II or more) were excluded. RESULTS: A total of 103 VLBW neonates were enrolled, in whom 1178 doppler measurements were recorded. The mean (SD) peak systolic velocity, end diastolic velocity and mean velocity (cm/s) in ACA increased from 26.53 (8.56) to 51.35 (9.36), 9.22 (2.91) to 13.9 (3.24) and 17.75 (3.97) to 25.84 (3.27) respectively from 2 to 8 hours to day 28 of life. In MCA and BA also, CBF velocities increased with post-natal age. CONCLUSION: We report normative data of CBF velocities in VLBW neonates in first 28 days of life.

Comparative Assessment of Herbal Mouthwash with Chlorhexidine on Plaque Accumulation, Gingivitis, and Salivary Streptococcus mutans Growth.

Objectives: The goal of the current study was to assess the effectiveness of a commercially available herbal mouthwash with chlorhexidine on the number of Streptococcus mutans in the saliva, the condition of the gingival tissue, and plaque development. Materials and Methods: Twenty-two adults in all, ranging in age from 20 to 30, were divided into two groups at random. Throughout the 14-day clinical trial, Group A (11) and Group B (11) received 10 mL of test herbal mouthwash and chlorhexidine, respectively. Result: Herbal mouthwash was discovered to be equally as effective as chlorhexidine in lowering the salivary mutans streptococci count and in affecting plaque and gingival scores. Conclusion: Alternatives from the herbal world may show to be a reliable and secure therapy option.

Study of biomarkers p53, Ki-67, Bcl-2, and VEGF in pterygium.

PURPOSE: To study the biomarkers present in primary pterygium samples of patients of Indian ethnicity and compare it with the samples obtained from the unaffected conjunctiva of the same eye. METHODS: A prospective case-control study of 17 eyes in patients above 10 years of age with primary pterygium who underwent pterygium excision using limbal conjunctival autograft technique. The pterygium samples (cases) and conjunctival samples (controls) were sent for immunohistochemical (IHC) staining for the following biomarkers: p53, Bcl-2, Ki-67, and vascular endothelial growth factor (VEGF). RESULT: The immunohistochemistry of the samples and the controls revealed p53 positivity in 47.05% of pterygium samples and 29.4% of controls ( P < 0.587). Nine cases each in pterygium and control samples were positive for Ki-67 expression. Differences in the staining pattern between the two groups were not statistically significant ( P < 1.000). Bcl-2 positivity was seen in 10 pterygium samples (58.8%) and 12 controls (70.5%), with no statistical difference between the two groups ( P < 0.455). VEGF expression was seen in both epithelial and endothelial cells of the samples and controls, with no statistical difference between the two groups, with P = 1.000 for the epithelial staining and P = 0.637 for endothelial staining. CONCLUSION: The expression of biomarkers was comparable in both groups. We conclude that pterygium, against common belief, might not be a localized disease process but a global ocular phenomenon where the apparently healthy tissue also has some ongoing disease process at a molecular level.

Staphylococcus hemolyticus: The Most Common and Resistant Coagulase-Negative Staphylococcus Species Causing Bacteremia in North India.

INTRODUCTION: Coagulase-negative Staphylococcus (CoNS) species are normal skin commensals but may also cause bacteremia. Therefore, isolating a CoNS species on blood culture often leads to a diagnostic dilemma about whether to consider the isolate as a true pathogen or not. This study was done to understand the distribution of various CoNS species in bloodstream infections, determine their antibiotic resistance patterns, and identify possible risk factors and patient outcomes in hospital settings. MATERIALS AND METHODS: Inpatients with confirmed bacteremia defined as isolation of the same CoNS species with similar antibiograms from paired blood culture bottles, which were obtained from patients with at least clinical evidence of infection, were included. The isolates obtained were studied for CoNS species distribution and antibiotic resistance patterns, and the corresponding patients were assessed for possible risk factors and outcomes. RESULTS: A total of 170 CoNS isolates obtained from 85 patients were analyzed. Staphylococcus haemolyticus (S. haemolyticus)(90, 52.9%) was the most common species isolated, and it was also the most resistant of all, followed by S. hominis (50, 29.4%), S. epidermidis (26, 15.3%), S. lentus (2,1.2%), and S. succinus (2,1.2%). S. haemolyticus and S. hominis were significantly more isolated from patients aged 18-60 years and >60 years, respectively. Methicillin-resistant (MR)-CoNS (68.8%) were significantly more resistant than methicillin-sensitive (MS)-CoNS (31.2%) to certain antibiotics, and none were resistant to vancomycin, linezolid, or teicoplanin. Mortality occurred in 17.6% of patients, which was most commonly associated with S. haemolyticus infection. CONCLUSION: Age-specific predisposition of CoNS species, high rates of methicillin resistance, and mortality in CoNS bacteremia are highlights of this study. To our knowledge, we are the first to study the age-related association of CoNS species.

High frequency of isoniazid and fluroquinolone resistance among patients with rifampicin sensitive tuberculosis.

This study was done to determine frequency of isoniazid (INH) and fluoroquinolones FQ resistance among rifampicin sensitive strains of Mycobacterium tuberculosis and to study their mutation patterns. Retrospective analysis was done for samples with M. tuberculosis detected by Cartridge based NAAT (CBNAAT). They were tested sequentially by first line (FL) and second line - line probe assay (SL-LPA) depending on their drug resistance pattern and following diagnostic algorithm. Total 9722 (74.1 %) of 13124 NAAT positive samples were sensitive for rifampicin. On FL-LPA, 833 (8.6 %) were resistant to INH and of which 110 (13.2 %) were also resistant to FQ by SL-LPA. Most common mutations observed for INH resistance were katG S315T1 mutation in 615 (97.3 %) strains, inhA C15T mutation in 174 (86.6 %) strains and for FQ resistance were gyrA D94G mutation in 46 (41.8 %) strains. Heteroresistance, inferred mutations, combination of mutations and unique mutations were also observed in all genes.

Advocating, Unlearning, Tearing It All Down? How BIPOC Young Adults with Mental Health Concerns Perceive and Engage with the Media.

The COVID-19 pandemic has increased the salience of issues related to social justice, mental health, and health equity. During this time, the media have been instrumental in amplifying social movements but also in spreading mis/disinformation, violence, and hatred. Among communities who have been affected heavily during this time are Black, Indigenous, and People of Color (BIPOC) young adults who have a mental illness/significant mental health concerns. As frequent engagers and creators of media, their perspectives are critical to informing what aspects of the media should be leveraged to promote the health, well-being, and safety of people who have been marginalized. Thus, our study asks: How do BIPOC young adults (in the United States) who have a behavioral health diagnosis or significant mental health concerns perceive the media? Guided by intersectionality and grounded theory, we conducted in-depth interviews with 20 participants. The results reveal participants' 1) deep recognition of how the media uphold oppressive structures and 2) identification of ways that people have used and can use the media to fight those same structures. Researchers and media leaders should listen to, center the experiences of, collaborate with, and follow the lead of people who have been marginalized. This is critical to transforming the media such that they promote the well-being of all people.

Super-resolution Reflection Microscopy via Absorbance Modulation.

In recent years, fluorescence microscopy has been revolutionized. Reversible switching of fluorophores has enabled circumventing the limits imposed by diffraction. Thus, resolution down to the molecular scale became possible. However, to the best of our knowledge, the application of the principles underlying super-resolution fluorescence microscopy to reflection microscopy has not been experimentally demonstrated. Here, we present the first evidence that this is indeed possible. A layer of photochromic molecules referred to as the absorbance modulation layer (AML) is applied to a sample under investigation. The AML-coated sample is then sequentially illuminated with a one-dimensional (1D) focal intensity distribution (similar to the transverse laser mode TEM01) at wavelength λ1 = 325 nm to create a subwavelength aperture within the AML, followed by illumination with a Gaussian focal spot at λ2 = 633 nm for high-resolution imaging. Using this method, called absorbance modulation imaging (AMI) in reflection, we demonstrate a 2.4-fold resolution enhancement over the diffraction limit for a numerical aperture (NA) of 0.65 and wavelength (λ) of 633 nm.

Determination of a cut-off value for the serological diagnosis of scrub typhus by detecting anti-Orientia tsutsugamushi immunoglobulin M.

Background & objectives: The diagnosis of scrub typhus (ST) is usually done using enzyme-linked immunosorbent assay (ELISA) due to its ease of performance and reading objectivity. The cut-off value for ELISA needs to be calculated for each geographical location as it depends on zonal endemicity of the disease. This study was, therefore, undertaken to calculate the pan-India cut-off for anti-Orientia tsutsugamushi (OT) immunoglobulin M (IgM) by ELISA. Methods: Samples from cases (cases of ST) and controls (voluntary, consenting, healthy adults) were collected by a network of 29 laboratories across India and tested for anti-OT IgM by immunofluorescence assay (IFA), the considered gold standard test. These samples were retested by ELISA for anti-OT IgM and their optical densities (ODs) were used for cut-off estimation by receiver operating characteristic (ROC) curve. Results: Anti-OT IgM ELISA ODs from 273 controls and 136 cases were used for the cut-off estimation. The ODs of the anti-OT IgM ELISA on healthy individuals and those of confirmed ST cases ranged from 0.1 to 0.75 and 0.5 to 4.718, respectively. ROC curve-based cut-off for ELISA was calculated as 0.554 at a sensitivity of 95.2 per cent and specificity of 95.1 per cent. A value of >1 was noted to have a specificity of 100 per cent in diagnosing ST. Interpretation & conclusions: The cut-off calculated for India was similar to the previous cut-off that was used until now.

Repeat deep anterior lamellar keratoplasty (DALK) for failed primary DALK.

Purpose: To evaluate the effectiveness of repeat deep anterior lamellar keratoplasty (DALK) in patients of previous failed DALK. Methods: : A retrospective analysis of records of seven patients who had undergone repeat DALK following the failure of the primary DALK was done. The indications for repeat surgery, time elapsed since the first surgery, and pre- & postoperative best-corrected visual acuity (BCVA) were noted for all the patients. Results: The follow-up period ranged between one- to four-year post repeat DALK. The indication of primary DALK was keratoconus with vernal keratoconjunctivitis (VKC) (n = 3), corneal amyloidosis (n = 2), Salzman nodular keratopathy (n = 1), and healed keratitis (n = 1). The need for repeat surgery arose when the BSCVA dropped to less than 20/200. The time interval elapsed since the first surgery ranged from two months to four years. Postoperatively, the BSCVA improved from 20/120 to 20/30 at the end of one-year post repeat DALK in all except one patient. All regrafts were clear at the most recent examination, performed after a mean period of 18 months after the secondary graft. No complication was encountered during the resurgery. The dissection of the host bed was easier in the second surgery owing to weaker adhesions. Conclusion: The prognosis for repeat DALK for failed DALK is excellent, and the outcomes of secondary grafts were comparable to those of primary DALK grafts. Re DALK offers the advantage of an easier dissection and lower chances of graft rejection compared to penetrating keratoplasty.

Application of a Hermite-based measure of non-Gaussianity to normality tests and independent component analysis.

In the analysis of neural data, measures of non-Gaussianity are generally applied in two ways: as tests of normality for validating model assumptions and as Independent Component Analysis (ICA) contrast functions for separating non-Gaussian signals. Consequently, there is a wide range of methods for both applications, but they all have trade-offs. We propose a new strategy that, in contrast to previous methods, directly approximates the shape of a distribution via Hermite functions. Applicability as a normality test was evaluated via its sensitivity to non-Gaussianity for three families of distributions that deviate from a Gaussian distribution in different ways (modes, tails, and asymmetry). Applicability as an ICA contrast function was evaluated through its ability to extract non-Gaussian signals in simple multi-dimensional distributions, and to remove artifacts from simulated electroencephalographic datasets. The measure has advantages as a normality test and, for ICA, for heavy-tailed and asymmetric distributions with small sample sizes. For other distributions and large datasets, it performs comparably to existing methods. Compared to standard normality tests, the new method performs better for certain types of distributions. Compared to contrast functions of a standard ICA package, the new method has advantages but its utility for ICA is more limited. This highlights that even though both applications-normality tests and ICA-require a measure of deviation from normality, strategies that are advantageous in one application may not be advantageous in the other. Here, the new method has broad merits as a normality test but only limited advantages for ICA.

Low-level language processing in brain-injured patients.

Assessing cognitive function-especially language processing-in severely brain-injured patients is critical for prognostication, care, and development of communication devices (e.g. brain-computer interfaces). In patients with diminished motor function, language processing has been probed using EEG measures of command-following in motor imagery tasks. While such tests eliminate the need for motor response, they require sustained attention. However, passive listening tasks, with an EEG response measure can reduce both motor and attentional demands. These considerations motivated the development of two assays of low-level language processing-identification of differential phoneme-class responses and tracking of the natural speech envelope. This cross-sectional study looks at a cohort of 26 severely brain-injured patient subjects and 10 healthy controls. Patients' level of function was assessed via the coma recovery scale-revised at the bedside. Patients were also tested for command-following via EEG and/or MRI assays of motor imagery. For the present investigation, EEG was recorded while presenting a 148 s audio clip of Alice in Wonderland. Time-locked EEG responses to phoneme classes were extracted and compared to determine a differential phoneme-class response. Tracking of the natural speech envelope was assessed from the same recordings by cross-correlating the EEG response with the speech envelope. In healthy controls, the dynamics of the two measures were temporally similar but spatially different: a central parieto-occipital component of differential phoneme-class response was absent in the natural speech envelope response. The differential phoneme-class response was present in all patient subjects, including the six classified as vegetative state/unresponsive wakefulness syndrome by behavioural assessment. However, patient subjects with evidence of language processing either by behavioural assessment or motor imagery tests had an early bilateral response in the first 50 ms that was lacking in patient subjects without any evidence of language processing. The natural speech envelope tracking response was also present in all patient subjects and responses in the first 100 ms distinguished patient subjects with evidence of language processing. Specifically, patient subjects with evidence of language processing had a more global response in the first 100 ms whereas those without evidence of language processing had a frontopolar response in that period. In summary, we developed two passive EEG-based methods to probe low-level language processing in severely brain-injured patients. In our cohort, both assays showed a difference between patient subjects with evidence of command-following and those with no evidence of command-following: a more prominent early bilateral response component.

Primary Intracranial Alveolar Soft Part Sarcoma: A Report of 3 Cases.

Alveolar soft part sarcoma (ASPS) commonly involves extremities and head and neck regions. Primary intracranial ASPS is rare. We report a series of 3 primary intracranial ASPS. These were not suspected clinically and histopathology with immunohistochemistry proved to be diagnostic in all 3 tumors.

Tumor infiltrating lymphocytes profile and response in neoadjuvant chemotherapy-treated triple-negative breast carcinoma patients.

Background: Triple negative breast carcinoma (TNBC) has the highest mortality among all the breast carcinoma subtypes, but paradoxically, it shows the best response to neoadjuvant chemotherapy (NACT). Tumor infiltrating lymphocytes (TIL) density has been shown to have prognostic significance in TNBC. However, there are limited data on TIL subpopulation and their association with response to NACT in TNBC. Materials and Methods: The study included 80 consecutive patients with TNBC prospectively diagnosed for two and half years, who underwent tru-cut biopsy before NACT, followed by subsequent definite surgical procedures. Global TIL profile and immunohistochemistry (IHC) analysis of CD3, CD4, CD8, CD20, and CD56 were done on all baseline tru-cut biopsies and post-NACT surgical specimens. Results: Almost half the patients were postmenopausal with a mean age of 45.89 ± 4.62 years. The majority had low CD3, low CD4, low CD56, low CD20, and high CD8 positivity in both pre- and post-NACT specimens. On multivariate analysis, low CD3, CD4, CD56 and CD 20 were established as independent predictor of poor pathologic response (PR). Low CD4 (adjusted odds ratio [OR]: 228.46) was associated with the highest OR for poor PR. Low CD8 was associated with significantly decreased odds of poor PR on univariate analysis (OR: 0.26), but it was not been established as an independent predictor of PR on multivariate logistic regression. NACT did not significantly alter the profile of TILs. Conclusions: TIL profile with low CD3, CD4, CD20, and CD56 expression predicts PR to NACT in TNBC and may thus help in prognostication of these patients.

Corneal aberrations, densitometry on scheimpflug imaging, and visual acuity after deep anterior lamellar keratoplasty.

PURPOSE: To evaluate visual acuity, corneal aberrations, and densitometry post deep anterior lamellar keratoplasty (DALK). METHODS: Twenty-five eyes of 25 patients who underwent DALK for corneal stromal diseases with normal endothelium were enrolled in a prospective interventional study. Evaluation for corneal aberrations and densitometry postoperatively was performed using Scheimpflug imaging at 6 months. Anterior Segment Optical Coherence Tomography (ASOCT) was done for host-graft morphology. Correlation analysis of postoperative best-corrected visual acuity with corneal aberrations and corneal densitometry (CD) was performed. RESULTS: Big bubble deep anterior lamellar keratoplasty (BB-DALK) was successful in 18 patients, while 7 eyes underwent manual dissection due to failed BB (Big Bubble) technique. The mean BSCVA was 1.35 ± 0.46 logMAR units pre-operatively which improved to 0.36 ± 0.19 logMAR units post DALK at 6 months (P < 0.001). The mean Root Mean Square (RMS) (µm) of the magnitude of total corneal aberrations, HOA and LOA from central 6mm zone of cornea were 14.6 ± 9.2, 4.7 ± 2.9, and 13.8 ± 8.8, respectively. The mean anterior (120µm), central and posterior corneal (60µm) densitometry in 0-2mm zone were 25.8 ± 7.7 GSU, 18.7 ± 4.16 GSU and 10.09 ± 3.9 GSU respectively. A statistically significant correlation of postoperative residual stromal bed thickness, total corneal HOA, LOA, total corneal aberrations, HOA front and posterior corneal densitometry (0-2mm zone) with postoperative BSCVA was found. CONCLUSION: Though there is a significant improvement in BSCVA after DALK, the quality of vision in terms of aberrometry and densitometry is suboptimal, especially in cases undergoing manual DALK compared to those undergoing big bubble formation.