Patterns of suicide and self-harm in Pakistan: a retrospective descriptive study protocol

Introduction Suicide is a major global public health problem. Low-income and middle-income countries contribute 78% of all suicidal deaths. Pakistan, a South Asian country, lacks official statistics on suicides at national level. Statistics on suicide are neither collected nationally nor published in the annual national morbidity and mortality surveys. Medicolegal reports on suicides and self-harm are extremely rich and important source of information but greatly underused in Pakistan. We aim to examine the patterns of suicides and self-harm retrospectively in patients who were registered with medicolegal centres (MLCs) in Karachi, during the period January 2017 to December 2021. Methods and analysis Using retrospective descriptive design, the data will be collected from the medical records maintained at the main office of the Karachi police surgeon. Data from all nine MLCs of Karachi are collated and stored at the main office of Police surgeon. Information on suicide and self-harm cases will be extracted from records of all MLCs. The data will be collected using structured proforma and it will be analysed using descriptive and inferential analysis. Ethics and dissemination The study was approved for exemption from Aga Khan University, Ethical Review Committee. The findings of the study will be disseminated by conducting seminars for healthcare professionals and stakeholders including psychiatrists, psychologists, counsellors, medicolegal officers, police surgeons, mental health nurses, general and public health physicians and policy makers. Findings will be published in local and international peer-reviewed scientific journals

Chemical Genetic Analysis and Functional Characterization of Staphylococcal Wall Teichoic Acid 2-Epimerases Reveals Unconventional Antibiotic Drug Targets

Here we describe a chemical biology strategy performed in Staphylococcus aureus and Staphylococcus epidermidis to identify MnaA, a 2-epimerase that we demonstrate interconverts UDP-GlcNAc and UDP-ManNAc to modulate substrate levels of TarO and TarA wall teichoic acid (WTA) biosynthesis enzymes. Genetic inactivation of mnaA results in complete loss of WTA and dramatic in vitro β-lactam hypersensitivity in methicillin-resistant S. aureus (MRSA) and S. epidermidis (MRSE). Likewise, the β-lactam antibiotic imipenem exhibits restored bactericidal activity against mnaA mutants in vitro and concomitant efficacy against 2-epimerase defective strains in a mouse thigh model of MRSA and MRSE infection. Interestingly, whereas MnaA serves as the sole 2-epimerase required for WTA biosynthesis in S. epidermidis, MnaA and Cap5P provide compensatory WTA functional roles in S. aureus. We also demonstrate that MnaA and other enzymes of WTA biosynthesis are required for biofilm formation in MRSA and MRSE. We further determine the 1.9Å crystal structure of S. aureus MnaA and identify critical residues for enzymatic dimerization, stability, and substrate binding. Finally, the natural product antibiotic tunicamycin is shown to physically bind MnaA and Cap5P and inhibit 2-epimerase activity, demonstrating that it inhibits a previously unanticipated step in WTA biosynthesis. In summary, MnaA serves as a new Staphylococcal antibiotic target with cognate inhibitors predicted to possess dual therapeutic benefit: as combination agents to restore β-lactam efficacy against MRSA and MRSE and as non-bioactive prophylactic agents to prevent Staphylococcal biofilm formation.publishe

COMPARATIVE ANALGESIC STUDIES OF LEAF AND STEM BARK OF CALOPHYLLUM INOPHYLLUM IN SWISS ALBINO MICE

Objectives: The aim of the present study was to perform acute oral toxicity studies and to compare and evaluate the analgesic effect of ethanolic extracts of leaf and stem bark of Calophyllum inophyllum on Swiss albino mice. Methods: Eddy's hotplate method and acetic acid induced writhing were implemented to determine analgesic properties of the extracts. Both the extracts were administered orally. Acute oral toxicity studies were conducted using the OECD guidelines 423 Annexure – 2d. Results: The results indicate the mortality was not observed during the acute oral toxicity studies and maximum safe does was determined. The analgesic effect of the extracts showed significant dose dependent effect (100 mg/kg b.w and 200 mg/kg b.w) on both models of algesia i.e., Eddy's hotplate method and acetic acid induced writhing. Conclusion: The comparative studies between the leaf and stem bark of Calophyllum inophyllum suggests that Calophyllum inophyllum leaf extract showed more activity compared to Calophyllum inophyllum stem bark extract

Patterns of suicide and self-harm in Pakistan: A retrospective descriptive study protocol

Introduction: Suicide is a major global public health problem. Low-income and middle-income countries contribute 78% of all suicidal deaths. Pakistan, a South Asian country, lacks official statistics on suicides at national level. Statistics on suicide are neither collected nationally nor published in the annual national morbidity and mortality surveys. Medicolegal reports on suicides and self-harm are extremely rich and important source of information but greatly underused in Pakistan. We aim to examine the patterns of suicides and self-harm retrospectively in patients who were registered with medicolegal centres (MLCs) in Karachi, during the period January 2017 to December 2021.Methods and analysis: Using retrospective descriptive design, the data will be collected from the medical records maintained at the main office of the Karachi police surgeon. Data from all nine MLCs of Karachi are collated and stored at the main office of Police surgeon. Information on suicide and self-harm cases will be extracted from records of all MLCs. The data will be collected using structured proforma and it will be analysed using descriptive and inferential analysis.Ethics and dissemination: The study was approved for exemption from Aga Khan University, Ethical Review Committee. The findings of the study will be disseminated by conducting seminars for healthcare professionals and stakeholders including psychiatrists, psychologists, counsellors, medicolegal officers, police surgeons, mental health nurses, general and public health physicians and policy makers. Findings will be published in local and international peer-reviewed scientific journals

Effects of feed additives on chicken growth and their residues in meat instigating deleterious consequences on the liver health of consumers - A prospective human study

The liver is a multifunctional organ that metabolizes all forms of diets, and food ingredients can have beneficial or detrimental effects on liver cells during such a process. As we have already studied the components of chicken feed previously, the purpose of this study was to observe the effects of chicken meat consumption on liver cells of rats. One hundred and twenty female Albino Wistar rats were randomly divided into four groups and separately for standard chow rat, commercial chicken feed, commercial chicken meat and organic chicken meat over a six-week period. Plasma levels of tumor necrosis factor alpha (TNF-α) and alpha-fetoprotein (AFP) were estimated before and after the administration of different meals. After the experiment, the liver samples were weighed and histopathologically evaluated using a knodell score to assess portal hypertension and liver damage. Animals fed with commercial chicken meat and feed for six weeks showed development of inflammation, necrosis, apoptosis and cirrhosis on histopathological examination of the liver, and had raised plasma TNF-α and AFP-levels. This study shows that damage to liver cells is caused by consumption of commercial feed for chickens and their meat which is considered safe for human consumption

Whole-genome sequencing of L638^R mutants.

<p>Heat map summary of all non-synonymous mutations identified by Illumina-based whole-genome sequencing (100X genome coverage) of L638^R mutants in MRSE CLB26329 (A) or MRSA COL (B). Red, non-synonymous mutation; grey, no change versus parental genome sequence; yellow, non-synonymous mutations in genes other than <i>mnaA</i>. Genome position, base pair change, and resulting amino acid residue substitution are highlighted. Note: with only one exception (<i>Δcap5P mnaA</i>_<i>Sa</i>^<i>D281Y</i>), no additional non-synonymous mutations besides the indicated <i>mnaA</i> mutation were identified in each of the drug resistant strains examined.</p

Genetic inactivation of <i>mnaA</i> in methicillin-resistant <i>Staphylococci</i> restores β-lactam susceptibility.

<p>Genetic inactivation of <i>mnaA</i> in methicillin-resistant <i>Staphylococci</i> restores β-lactam susceptibility.</p

MRSA and MRSE MnaA LOF mutants are highly susceptible to imipenem in a murine thigh infection.

<p>Immune-suppressed CD-1 mice (5 per group) were challenged intramuscularly with the parental MRSA COL strain, MRSA <i>Δcap5P</i>, or MRSA <i>Δcap5P mnaA</i>_<i>Sa</i> LOF mutants (A) or with the parental MRSE strain versus <i>mnaA</i>_<i>Se</i>, <i>tarO</i>_<i>Se</i> and <i>tarA</i>_<i>Se</i> LOF mutants (B) and treated three times daily (TID) with imipenem (IPM). Thighs were harvested at 24hrs, homogenized and plated to determine CFU per thigh. (A) Restored efficacy of IPM (10 mg kg^-1) against MRSA <i>Δcap5P mnaA</i>_<i>Sa</i>^<i>P12L</i>, <i>Δcap5P mnaA</i>_<i>Sa</i>^<i>Y194</i>*, and <i>Δcap5P mnaA</i>_<i>Sa</i>^<i>D281Y</i>. Following IPM treatment, bacterial burden amongst mice infected with <i>Δcap5P mnaA</i>_<i>Sa</i>^<i>P12L</i>, <i>Δcap5P mnaA</i>_<i>Sa</i>^<i>Y194</i>*, and <i>Δcap5P mnaA</i>_<i>Sa</i>^<i>D281Y</i> is reduced approximately 2–3 log at 24 hrs versus those infected with MRSA COL or <i>Δcap5P</i> controls. * p<0.01 versus parent at 24 hr; $ p<0.05 versus respective 24 hr vehicle. (B) Restored efficacy of IPM (2.5 mg kg^-1) against MRSE <i>mnaA</i>, <i>tarO</i>, and <i>tarA</i> LOF mutants. Reduction in bacterial burden of mice infected with the <i>mnaA</i>_<i>Se</i>^<i>G171D</i> is comparable to those infected with <i>tarO</i>_<i>Se</i>^<i>G84</i>* or <i>tarA</i>_<i>Se</i>^<i>G129R</i> mutants, yielding an approximate 3 log reduction in 24 hr IPM treatment versus the wild-type control. Note, as MRSE CLB26329 is more susceptible to IPM than MRSA COL, its dose was reduced to 4-fold versus the MRSA efficacy study (A).</p

Tunicamycin inhibits MnaA in a concentration-dependent manner.

<p>Representative electropherograms of the MnaA-catalyzed conversion of UDP-ManNAc to UDP-GlcNAc in the absence of tunicamycin (A), in the presence of 100 μM (B) and 200 μM tunicamycin (C). Substrate concentration is 100 μM. Peaks: <b>1</b> UDP-ManNAc; <b>2</b> UDP-GlcNAc.</p

Biophysical studies demonstrate MnaA and Cap5P bind tunicamycin.

<p>(A, D) 600 MHz 1H NMR spectra of 15 μM tunicamycin. (B, E) 1H NMR STD spectra of 15 μM tunicamycin without 2-epimerase. (C) 1H NMR STD spectra of 15 μM tunicamycin in presence of 5 μM MnaA. (F) 1H NMR STD spectra of 15 μM tunicamycin in presence of 5 μM Cap5P. Saturation of the protein was achieved with a Gaussian pulse cascade resulting in a total saturation time of 3s. The protein resonances were saturated at 100 Hz and the off resonance was set to -120 ppm. Tunicamycin-specific peaks in NMR STD spectra were only obtained in the presence of MnaA or Cap5P. (G) Structure of tunicamycin.</p