Comparison of changes in retinal nerve fiber layer thickness and intraocular pressure between glaucoma and non-glaucoma patients after phacoemulsification.

Objective: To compare the changes in Retinal Nerve Fiber Layer (RNFL) thickness and Intraocular Pressure (IOP) after phacoemulsification in patients with glaucoma versus without glaucoma. Methods: This was a quasi-experimental study, done at Lahore General Hospital, from January 2021 to December 2021. Patients with senile cataract were divided into two groups. One group included 40 patients, with ≥40 years of age and diagnosed with primary open angle glaucoma compared with a second group of 40 age-matched controls without glaucoma, undergoing phacoemulsification and Intraocular Lens Implantation (IOL). Goldman Applanation Tonometer (GAT) and Spectral Domain Optical Coherence Tomography (SD-OCT) were used to record IOP and RNFL thickness, before surgery, at one week and one month after surgery. The results were compared with baseline readings and also between the two groups. Results: A total of 80 patients were included in the study. Mean age was 60.3±7.9 years with male to female ratio of 1:1. Mean change in RNFL thickness in glaucomatous eyes group and non-glaucomatous Eyes group from pre-operative baseline was 11.33±4.30µm and 4.08±2.59µm respectively after one month (p-value<0.001). Difference of mean change in RNFL thickness from baseline was statistically significant between both groups (p< 0.001). Difference of mean change in IOP from baseline at four weeks was statistically significant in the individual groups but between both groups was statistically insignificant (p= 0.234). Conclusion: Phacoemulsification results in increase in RNFL thickness and decrease in IOP, which are good prognostic factors in control of glaucoma. However, a mean change of IOP of 1mmHg in glaucoma patient does not affect management of glaucoma.

A clinical audit to evaluate antibiotic prescribing practice in pediatric patients admitted for enteric fever. Rationalizing antibiotic stewardship program.

To evaluate the antibiotic prescribing practice in pediatric patients for enteric fever, and to assess the need of developing and implementing the Antibiotic Stewardship Program (ASP) for the hospital. A prospective audit was completed in the pediatric ward of tertiary care hospital of Lahore for one year. Blood culture reports were collected from microbiology departed and clinical data were assessed regarding the choice of antibiotics, frequency, dosage and clinical outcome. All the statistics were analyzed using SPSS software and compared with the guidelines. Out of 157 cases hospitalized with suspicion of enteric fever, 137 cultures were positive for salmonella. Monotherapy of ceftriaxone (70%) was prescribed mostly as empirical therapy. About 20% of patients received a combination of antibiotics empirically. Susceptibility reports showed only 7 cases were of non-resistant typhoid, 15 multi-drug resistant and 115 extensively drug-resistant. Nearly 46% of patients were discharged earlier whose empirical therapy was changed either before or promptly after susceptibility reporting. Commonly used definitive antibiotics (32%) were a combination of azithromycin and meropenem. Inappropriate use of antibiotics was noted frequently as compared to the guidelines. However, recommendations themselves need to be reviewed as antibiotic resistance patterns are changing drastically.

Phytochemistry, antioxidant and antibacterial activities of fruit extracts of Myrsine africana L.

Myrsine africana L. a commonly consumed medicinal plant grows in forest of mountains region located at North East of Pakistan. In current study, the fruit extracts were chemically characterized and their bioactivities were determined. Higher quantity of total phenols, total flavonoids and tannins were obtained from methanolic fruit extracts. The HPLC analysis provided higher level of quercetin followed by rutin and p-coumaric acid. Whereas the GC-MS quantification had given significant level of ten saturated and unsaturated fatty acids and some of them were not reported earlier. In vitro study, lower cytotoxic behavior of fruit extracts but higher antioxidant values as well as higher zone of inhibition versus S. aureus, E. coli, K. pneumonia and B. subtilis and Mycobacterium tuberculosis were observed. The organic compounds found in fruit extracts of M. africana correlated well with its used in ethno medicines.

Effects of smartphone usage on psychological wellbeing of school going children in Lahore, Pakistan.

OBJECTIVE: To analyse the effects of smartphone usage on the psychological wellbeing of schoolgoing children. METHODS: The descriptive cross-sectional community household survey was conducted from January to May 2017 in all the 10 towns of Lahore, Pakistan and comprised school-going smartphone users. A self-administered structured questionnaire was used for data collection. Children who used smartphones for up to 2 hours a day were considered short-term users while children who used smartphones for more than 2 hours a day were considered long-term users. SPSS 21 was used for data analysis. RESULTS: Of the 2889 subjects, 1993(69%) were short-term users and 896(31%) were long-term users. Long-term smartphone usage was significantly associated with behavioural problems (p<0.05) as well as with psychological problems (p<0.05). CONCLUSIONS: Long-term smartphone usage was found significantly associated with behavioural and psychological problems among school-going children.

Mechanism of Interferon-Stimulated Gene Induction in HIV-1-Infected Macrophages.

Viruses manipulate the complex interferon and interferon-stimulated gene (ISG) system in different ways. We have previously shown that HIV inhibits type I and III interferons in its key target cells but directly stimulates a subset of >20 ISGs in macrophages and dendritic cells, many of which are antiviral. Here, we examine the mechanism of induction of ISGs and show this occurs in two phases. The first phase was transient (0 to 24 h postinfection [hpi]), induced mainly by extracellular vesicles and one of its component proteins, HSP90α, contained within the HIV inoculum. The second, dominant, and persistent phase (>48 hpi) was induced via newly transcribed HIV RNA and sensed via RIGI, as shown by the reduction in ISG expression after the knockdown of the RIGI adaptor, MAVS, by small interfering RNA (siRNA) and the inhibition of both the initiation and elongation of HIV transcription by short hairpin RNA (shRNA) transcriptional silencing. We further define the induction pathway, showing sequential HIV RNA stimulation via Tat, RIGI, MAVS, IRF1, and IRF7, also identified by siRNA knockdown. IRF1 also plays a key role in the first phase. We also show that the ISGs IFIT1 to -3 inhibit HIV production, measured as extracellular infectious virus. All induced antiviral ISGs probably lead to restriction of HIV replication in macrophages, contributing to a persistent, noncytopathic infection, while the inhibition of interferon facilitates spread to adjacent cells. Both may influence the size of macrophage HIV reservoirs in vivo Elucidating the mechanisms of ISG induction may help in devising immunotherapeutic strategies to limit the size of these reservoirs.IMPORTANCE HIV, like other viruses, manipulates the antiviral interferon and interferon-stimulated gene (ISG) system to facilitate its initial infection and establishment of viral reservoirs. HIV specifically inhibits all type I and III interferons in its target cells, including macrophages, dendritic cells, and T cells. It also induces a subset of over 20 ISGs of differing compositions in each cell target. This occurs in two temporal phases in macrophages. Extracellular vesicles contained within the inoculum induce the first, transient phase of ISGs. Newly transcribed HIV RNA induce the second, dominant ISG phase, and here, the full induction pathway is defined. Therefore, HIV nucleic acids, which are potent inducers of interferon and ISGs, are initially concealed, and antiviral ISGs are not fully induced until replication is well established. These antiviral ISGs may contribute to persistent infection in macrophages and to the establishment of viral reservoirs in vivo.

Determinants of patients preferring Complementary and Alternative medicine attending public hospitals in Lahore, Pakistan.

OBJECTIVE: To find the frequency and determinants of patients preferring complementary and alternative medicine over registered medical doctors.. METHODS: This cross-sectional study was conducted at Services Hospital, Mayo Hospital and Jinnah Hospital in Lahore, Pakistan, from June 8 to August 20, 2017.A pre-tested self-administered questionnaire was used to collect data. SPSS 20 was used for data analysis. . RESULTS: Of the 385 subjects, 200(51.9%) had visited at least one complementary and alternative medicine healer in their life. Besides, 166(83%) subjects confirmed positive outcome of such therapies. When asked about reasons behind their choice,227(59%)subjects mentioned cost effectiveness, 99(25.7%) better understanding, 131(34%) guidance about disease by such healers, 198(51.4%) harmless therapies, 198(51.4%) natural ingredients of medicines, 154(40%) accessibility, 161(41.8%) psychological satisfaction, 221(57.4%) said fewer appointments, 222(57.7%) said diagnosis without laboratory tests, 131(34%)family influence, 52(13.5%) had surgical fears, 101(26.2%) said faith in spiritual exercises, 63(16.4%) were addicted to complementary and alternative medicine products, 122(31.7%) said better communication) while183(47.5%) said sparing more time for consultation. CONCLUSIONS: There were a variety of beliefs and reasons behind patients preferring complementary and alternative medicine providers over medical doctors.

Flagellar motility plays important role in Biofilm formation of Bacillus cereus and Yersinia enterocolitica.

Bacteria live either independently as planktonic cells or in organized surface associated colonies called as biofilms. Biofilms play an important role in increased pathogenesis of bacteria and it is assumed that motility is one of the contributing factors towards biofilm initiation. This study was planned to identify the role of flagella in biofilm formation by constructing flagellated (wild type) and physically disrupted variants (non-motile). Total 10 clinical bacterial strains were isolated and characterized. Morphological and biochemical study identified these strains as Enterobacter spp., Pseudomonas spp., Yersinia spp., Escherichia spp., Salmonella spp., Proteus spp., Staphylococcus spp., Streptococcus spp., Lactobacillus spp. and Bacillus spp. Among all strains, two strains including Yersinia spp and Bacillus spp. showed higher antibiotic resistance, hence studied at molecular and physiological level. Biofilm formation capacity of strains was analyzed using three methods including Congo red assay, Test tube assay and Liquid-interface coverslip assay. Afterwards, flagellar disintegration was induced by blending and centrifugation for 5, 10 and 15 minutes. 16S rRNA sequencing showed two strains as Bacillus cereus and Yersinia enterocolitica. Both strains produced significant biofilm by all three above mentioned methods. A motility test of these blended variants showed partial/diminished motility with increased blending time. The significant loss in biofilm formation after 15 minutes blending confirmed the important flagellar contribution to the initiation of biofilm formation. This biofilm defect observed in flagella paralysed/minus variants presumably may be due to defects in attachments to surface at early stages. This study indicated that flagellar motility is crucial initially for surface attachment and subsequently for biofilm formation.

Automated prior knowledge-based quantification of neuronal patterns in the spinal cord of zebrafish.

MOTIVATION: To reliably assess the effects of unknown chemicals on the development of fluorescently labeled sensory-, moto- and interneuron populations in the spinal cord of zebrafish, automated data analysis is essential. RESULTS: For the evaluation of a high-throughput screen of a large chemical library, we developed a new method for the automated extraction of quantitative information from green fluorescent protein (eGFP) and red fluorescent protein (RFP) labeled spinal cord neurons in double-transgenic zebrafish embryos. The methodology comprises region of interest detection, intensity profiling with reference comparison and neuron distribution histograms. All methods were validated on a manually evaluated pilot study using a Notch inhibitor dose-response experiment. The automated evaluation showed superior performance to manual investigation regarding time consumption, information detail and reproducibility. AVAILABILITY AND IMPLEMENTATION: Being part of GNU General Public Licence (GNU-GPL) licensed open-source MATLAB toolbox Gait-CAD, an implementation of the presented methods is publicly available for download at http://sourceforge.net/projects/zebrafishimage/.

Occurrence of Virulence Genes among Methicillin-Resistant Staphylococcus aureus Isolated from Subclinical Bovine Mastitis.

The occurrence of Staphylococcus aureus-induced subclinical mastitis holds significant implications for public health. This specific microorganism possesses a wide array of pathogenic factors that enable it to adhere to, colonize, invade, and infect the host. The objective of the current study was to assess the prevalence of S. aureus, determine antimicrobial resistance patterns, and identify virulence genes of methicillin-resistant S. aureus (MRSA) strains responsible for subclinical mastitis in bovines. A total of 249 milk samples were collected from various farms in the district of Faisalabad. The presence of subclinical mastitis was assessed by using the California mastitis test. Positive milk samples (n = 100) were then subjected to standard microbiological techniques for isolation and identification of S. aureus. Antibiogram analysis was conducted by using the disc diffusion method to assess antimicrobial resistance. For the molecular detection of S. aureus and its virulence genes, the polymerase chain reaction (PCR) was performed with species-specific primers. The overall prevalence of S. aureus was found to be 40% (40/100), which was confirmed through molecular detection of the nuc gene in 40/40 (100%) of samples using PCR. Antimicrobial susceptibility tests indicated the highest susceptibility to vancomycin, sulfamethoxazole/trimethoprim, erythromycin, gentamicin, ciprofloxacin, and chloramphenicol, while the highest resistance rate was observed against tetracycline. Additionally, 30% of samples (12/40) tested positive for methicillin resistance. PCR analysis revealed that 100% of MRSA-tested isolates harbored the mecA and clfA genes. Furthermore, the MRSA isolates showed the presence of pvl, hla, hlb, sec, icaA, icaD, icaB, and icaC genes at rates of 92, 75, 67, 42, 42, 75, 8, and 25%, respectively. These findings underscore the need for stricter aseptic control in dairy farms to prevent disease transmission between animals and ensure the production of safe and uncontaminated food for human consumption.

Frequency and Severity of Thrombocytopenia in Neonatal Sepsis.

Background Neonatal sepsis includes numerous systemic illnesses such as septicemia, meningitis, urinary tract infections, and pneumonia. In developing countries, the major reason for neonatal mortality is septicemia, which accounts for almost 50% of overall deaths. Thrombocytopenia is one of the most common hematological problems during the neonatal period, affecting the majority of sufferers admitted to the neonatal intensive care unit (NICU). The aim of our study was to find the frequency of thrombocytopenia and its severity in neonates with sepsis. Methods The study was conducted at the Department of Hematology at Khyber Medical University, Peshawar, Pakistan. A total of 170 neonates with an age of fewer than 28 days, both genders, and positive blood cultures were included in the study using a non-probability consecutive sampling technique. Data was recorded in predesigned questionnaires after taking informed consent. Data were recorded and analyzed using SPSS version 26 (IBM Corporation, Armonk, NY, USA). Results Of the 170 neonates, 104 (61.2%) were males, with a mean age of 12.12±8.88 days. The majority of the babies 73 (42.9%) were in the age group of 0-7 days. Most of the neonates 72 (42.4%) were born via normal vaginal delivery (NVD). Of the neonates, 117 (68.82%) presented with fever, and 105 (61.76%) were reluctant to feed. Furthermore, 65.29% of the neonates had thrombocytopenia, of which 34 (20%) had mild, 43 (25.3%) had moderate, and 34 (20%) had severe thrombocytopenia. In neonates with positive blood culture, the platelet level was low (p<0.001). In the case of gram-negative organisms, the level of platelets was lower as compared to gram-positive organisms (p<0.001). Conclusion Sepsis is still a common cause of newborn thrombocytopenia. The fact that it is present in more than half of all culture-positive sepsis episodes indicates the severity of the condition. This condition is further defined by higher percentages of early-onset gram-negative septicemia compared to gram-positive sepsis.

Therapeutic Chemical Screen Identifies Phosphatase Inhibitors to Reconstitute PKB Phosphorylation and Cardiac Contractility in ILK-Deficient Zebrafish.

Patients with inherited dilated cardiomyopathy (DCM) often suffer from severe heart failure based on impaired cardiac contractility leading to increased morbidity and mortality. Integrin-linked kinase (ILK) as a part of the cardiac mechanical stretch sensor was found to be an essential genetic regulator of cardiac contractility. Integrin-linked kinase localizes to z-disks and costameres in vertebrate hearts and regulates the activity of the signaling molecule protein kinase B (PKB/Akt) by controlling its phosphorylation. Despite identification of several potential drug targets in the ILK signaling pathway, pharmacological treatment strategies to restore contractile function in ILK-dependent cardiomyopathies have not been established yet. In recent years, the zebrafish has emerged as a valuable experimental system to model human cardiomyopathies as well as a powerful tool for the straightforward high-throughput in vivo small compound screening of therapeutically active substances. Using the ILK deficient zebrafish heart failure mutant main squeeze (msq), which shows reduced PKB phosphorylation and thereby impaired cardiac contractile force, we identified here, in an automated small compound screen, the protein phosphatase inhibitors calyculin A and okadaic acid significantly restoring myocardial contractile function by reconstituting PKB phosphorylation in msq ILK-deficient zebrafish embryos.

Zebrafish biosensor for toxicant induced muscle hyperactivity.

Robust and sensitive detection systems are a crucial asset for risk management of chemicals, which are produced in increasing number and diversity. To establish an in vivo biosensor system with quantitative readout for potential toxicant effects on motor function, we generated a transgenic zebrafish line TgBAC(hspb11:GFP) which expresses a GFP reporter under the control of regulatory elements of the small heat shock protein hspb11. Spatiotemporal hspb11 transgene expression in the musculature and the notochord matched closely that of endogenous hspb11 expression. Exposure to substances that interfere with motor function induced a dose-dependent increase of GFP intensity beginning at sub-micromolar concentrations, while washout of the chemicals reduced the level of hspb11 transgene expression. Simultaneously, these toxicants induced muscle hyperactivity with increased calcium spike height and frequency. The hspb11 transgene up-regulation induced by either chemicals or heat shock was eliminated after co-application of the anaesthetic MS-222. TgBAC(hspb11:GFP) zebrafish embryos provide a quantitative measure of muscle hyperactivity and represent a robust whole organism system for detecting chemicals that affect motor function.

Dysferlin-mediated phosphatidylserine sorting engages macrophages in sarcolemma repair.

Failure to repair the sarcolemma leads to muscle cell death, depletion of stem cells and myopathy. Hence, membrane lesions are instantly sealed by a repair patch consisting of lipids and proteins. It has remained elusive how this patch is removed to restore cell membrane integrity. Here we examine sarcolemmal repair in live zebrafish embryos by real-time imaging. Macrophages remove the patch. Phosphatidylserine (PS), an 'eat-me' signal for macrophages, is rapidly sorted from adjacent sarcolemma to the repair patch in a Dysferlin (Dysf) dependent process in zebrafish and human cells. A previously unrecognized arginine-rich motif in Dysf is crucial for PS accumulation. It carries mutations in patients presenting with limb-girdle muscular dystrophy 2B. This underscores the relevance of this sequence and uncovers a novel pathophysiological mechanism underlying this class of myopathies. Our data show that membrane repair is a multi-tiered process involving immediate, cell-intrinsic mechanisms as well as myofiber/macrophage interactions.