Trimester-specific reference ranges for thyroid hormones of pregnant females at tertiary care hospitals in Lahore, Pakistan.

BACKGROUND: The significance of investigation for diagnosing and managing thyroid dysfunction in pregnant females has been extensively documented in the medical literature. This study aimed to determine trimester-specific reference ranges for thyroid-stimulating hormones (TSH), free T3 (FT3), and free T4 (FT4) in apparently healthy pregnant women attending tertiary care hospitals in Lahore. METHODS: This cross-sectional study was conducted at two tertiary care Hospitals in Lahore, Pakistan. In this multi-centric study, 500 pregnant females were initially enrolled from September 2019 to December 2019 who fulfilled the inclusion criteria. For measurement of serum FT3, FT4, thyroid stimulating hormone (TSH), anti-thyroid peroxidase (anti-TPO), and thyroglobulin antibodies, 5 ml of the blood sample was drawn, under aseptic conditions, from each subject using Maglumi 800 chemiluminescence immunoassay (CLIA) system. RESULTS: Out of 500 subjects, 23 subjects with positive anti-TPO, 19 subjects with anti-TG antibodies, and 12 subjects due to less volume of serum yielded from whole blood (serum less than 3 ml) were excluded from the analysis. Ten samples were hemolyzed and not included in the analysis. A total of 436 samples were examined for analysis. Of the remaining 436 subjects, 133 (30.5%) were from 1st trimester, 153 (35.1%) from 2nd trimester, and 150 (34.4%) from 3rd trimester. As the data were non-normal, the 2.5th, 50th, and 97.5th percentiles were calculated to express each group's results. Trimester specific range of TSH 0.168-4.294, 0.258-4.584 and 0.341-4.625 mIU/mL, FT31.857-4.408, 1.958-4.621 and 2.025-4.821 pmol/L and FT4 8.815-18.006, 8.306-17.341 and 7.402-17.292 pmol/L. CONCLUSION: In this study, we established a trimester-specific reference range for our local population's thyroid function test. The results of this study have complemented the results of previous studies.

Frequency of high bleeding risk in patients undergoing percutaneous coronary intervention.

OBJECTIVE: To assess the prevalence of risk factors for bleeding and high bleeding risk in patients undergoing percutaneous coronary intervention. METHODS: The single-centre retrospective observational study was conducted at Ittefaq Hospital Lahore and comprised data of patients who underwent percutaneous coronary intervention from February 2018 to December 2019. Minor and major bleeding risk factors were identified on the basis of the consensus definition of the Academic Research Consortium. Patients with high bleeding risk were identified according to the consensus-based criteria of at least one major criterion or two minor criteria. Data was analysed using SPSS 20. RESULTS: Of the 385 patients, 280(72.7%) were males. The overall mean age was 57.9±11.9 years. The indication of procedure was acute coronary syndrome in 367(95%) patients. Of all the patients, 171(45%) had bleeding risk, with 94(24%) patients having a high bleeding risk. Of these, 60(15.6%) patients had high risk based on the presence of at least one major criterion and 34(8.8%) patients because of the presence of two or more minor criteria. Patients with high bleeding risk were more likely to be older and female with more co-morbidities (p<0.05). CONCLUSIONS: Almost half of the patients undergoing percutaneous coronary intervention were found to have at least one bleeding risk factor, and one in four patients had high bleeding risk.

Synthesis, in-vitro, in-vivo anti-inflammatory activities and molecular docking studies of acyl and salicylic acid hydrazide derivatives.

Over the course of time several drugs have been synthesized and are available in market for the treatment of inflammation. However, they were unable to cure effectively and associated with side effects. To effectively deal with such diseases, heterocycles and their derivatives have gained their special position. For this reason 1,3,4-oxadiazole (15-16), 1,2,4-triazole (17-18), Schiff base (19-24) and 3,5-disubstituted pyrazole (25) derivatives were synthesized starting from salicylic acid and acyl acid hydrazides (12-14) as COX-1 and COX-2 inhibitors. In vivo anti-inflammatory activities were also tested by carrageenan-induced mice paw edema against albino mice of any sex. Structures of all the synthesized compounds were confirmed by FT-IR and 1H NMR analysis. Schiff base derivative of 4-amiontirazole (24) with IC50 value of 1.76 ± 0.05 (COX-2) and 117.8 ± 2.59 emerged as potent COX-2 inhibitor. Furthermore, we also performed in-vivo anti-inflammatory investigations by using carrageenan induced paw edema test. From in-vivo anti-inflammatory activities, it was found that after 1 h the maximum percentage inhibition 15.8% was observed by compound 14 which is comparable with that of the standard drug followed by the compound 18 with percentage inhibition of 10.5%. After 3 h, the maximum percentage inhibition was observed by compound 18 with 22.2% and compound 14 with 16.7%. After 5 h the maximum percentage inhibition was observed by compound 18 with 29.4% followed by compound 16 with 23.5%. We further explore the mechanism of the inhibition by using docking simulations. Docking studies revealed that the selective COX-2 inhibitors established interactions with additional COX-2 enzyme pocket residues.

Bisthioureas of pimelic acid and 4-methylsalicylic acid derivatives as selective inhibitors of tissue-nonspecific alkaline phosphatase (TNAP) and intestinal alkaline phosphatase (IAP): Synthesis and molecular docking studies.

Alkaline phosphatases (ALPs) are membrane bound metalloenzymes, distributed all over the body. Recent studies have revealed that by targeting ALPs can lead towards the treatment of many deadliest diseases including cardiac, cancerous and brain diseases. Thioureas and their derivatives are of considerable significance and are privileged scaffolds in medicinal chemistry. They show a wide range of pharmacological activities such as antibacterial, antiparasitic, anti-inflammatory and antioxidants etc. On the other hand, salicylic acid and its derivatives are known for its broad spectrum of activities. The work presented comprises of synthesis of N-acyl-N'-aryl substituted bisthioureas of pimelic acid (1-7) and 3,5-dimethyl pyrazole (11), 1-aroyl-3-aryl thiourea (12) and 1,3,4-oxadiazole (13) derivatives of 4-methyl salicylic acid. Structures of all the synthesized compounds were characterized by FT-IR and 1H NMR spectroscopic analysis. Synthesized compounds were evaluated for their alkaline phosphatases inhibition potential and exhibited high potency as well as selectivity towards h-TNAP and h-IAP. Compound 7 and 12 which were the bisthiourea derivative of pimmelic acid and thiourea derivative of 4-methyl salicylic acid, respectively, showed excellent selectivity against h-TNAP and h-IAP, respectively.

Resveratrol regulates hyperglycemia-induced modulations in experimental diabetic animal model.

Type 2 diabetes mellitus (T2DM) is a metabolic disorder that is associated with variable degrees of glucose intolerance, impaired insulin secretion, insulin resistance and increased hepatic glucose production. The aim of present study was to investigate the therapeutic potentials of resveratrol (RSV) alone and/or in combination with vitamin-E (Vit-E) against hyperglycemia-induced modulations using experimentally alloxan-induced diabetic animal model. Alloxan was used to induce diabetes mellitus in white albino rats and metformin (MF) was used as standard anti-diabetic drug to compare the therapeutic potentials of RSV (alone and/or with Vit-E) by estimating the effect of treatment on glycemia, insulin resistance, liver and kidney function biomarkers, anti-oxidant status, and serum levels of calcium and magnesium. The results of present study indicate the RSV (P < 0.001) alone and/or in combination with Vit-E (P < 0.001) exhibited a highly significant therapeutic potentials by ameliorating the glycemia-induced modulations. Moreover, we also found that RSV in combination with Vit-E also exhibited a better therapeutic effects when compared with that of MF (P < 0.001) and Vit-E (P < 0.05), respectively. Hence, we conclude that RSV alone and/or in combination with Vit-E exhibit its significant therapeutic potentials against hyperglycemia-induced modulations in experimental diabetic animal model and may be one of the most exciting prospect for future treatment of T2DM.

Conformational analysis and geometry optimization of apomorphine as an Anti-parkinsonian agen.

Apomorphine, a dopamine D1/D2agonist, is an important drug of choice for the treatment of Parkinson's and related disorders. The present study was designed to perform the conformational analysis and geometry optimization of apomorphine. Resultant optimized structure corresponds to a substance as it is found in nature. This could be used for a variety of experimental and theoretical investigations especially in the field of pharmacokinetics. The results indicate that the best conformation of the molecule is present at minimum potential energy -88702.9595 kcal/mol. At this point molecule will be more active as histamine H1 receptor agonist.

Metabolic pathway analysis approach: identification of novel therapeutic target against methicillin resistant Staphylococcus aureus.

Multiple Drug Resistant (MDR) bacteria are no more inhibited by the front line antibiotics due to extreme resistance. Methicillin Resistant Staphylococcus aureus (MRSA) is one of the MDR pathogens notorious for its widespread infection around the world. The high resistance acquired by MRSA needs a serious concern and efforts should be carried out for the discovery of better therapeutics. With this aim, we designed a comparison of the metabolic pathways of the pathogen, MRSA strain 252 (MRSA252) with the human host (i.e., Homo sapiens) by using well-established in silico methods. We identified several metabolic pathways unique to MRSA (i.e., absent in the human host). Furthermore, a subtractive genomics analysis approach was applied for retrieval of proteins only from the unique metabolic pathways. Subsequently, proteins of unique MRSA pathways were compared with the host proteins. As a result, we have shortlisted few unique and essential proteins that could act as drug targets against MRSA. We further assessed the druggability potential of the shortlisted targets by comparing them with the DrugBank Database (DBD). The identified drug targets could be useful for an effective drug discovery phase. We also searched the sequences of unique as well as essential enzymes from MRSA in Protein Data Bank (PDB). We shortlisted at least 12 enzymes for which there was no corresponding deposition in PDB, reflecting that their crystal structures are yet to be solved! We selected Glutamate synthase out of those 12 enzymes owing to its participation in significant metabolic pathways of the pathogen e.g., Alanine, Aspartate, Glutamate and Nitrogen metabolism and its evident suitability as drug target among other MDR bacteria e.g., Mycobacteria. Due to the unavailability of any crystal structure of Glutamate synthase in PDB, we generated the 3D structure by homology modeling. The modeled structure was validated by multiple analysis tools. The active site of Glutamate synthase was identified by not only superimposing the template structure (PDB ID: 1E0A) over each other but also by the Parallel-ProBiS algorithm. The identified active site was further validated by cross-docking the co-crystallized ligand (2-oxoglutaric acid; AKG) of PDB ID: 1LLW. It was concluded that the comparative metabolic in silico analysis together with structure-based methods provides an effective approach for the identification of novel antibiotic targets against MRSA.

An exhaustive yet simple virtual screening campaign against Sortase A from multiple drug resistant Staphylococcus aureus.

Methicillin resistant Staphylococcus aureus (MRSA) is one of the challenging bacterial pathogen due to its acquired resistance to the ß lactam antibiotics. The Sortase A is an enzyme of Gram-positive bacteria including S. aureus to anchor surface proteins to the cell wall. Sortase A is well studied enzyme and considered as the drug target against MRSA. Sortase A plays active role in anchoring the virulence proteins on the cell wall of the Gram-positive bacteria. The inhibition of Sortase A activity results in the separation of S. aureus from the host cells and ultimately alleviation of the infection. Here, we adapted a structure-based virtual screening protocol which helped in identification of novel potential inhibitors of Sortase A. The protocol involved the docking of a chemical library of druglike compounds with the Sortase A binding site represented by multiple crystal structures. The compounds were ranked by multiple scoring functions and shortlisted for future experimental screening. The method resulted in shortlisting of three compounds as potential novel inhibitors of Sortase A out of a large chemical library. The high rankings of shortlisted compounds estimated by multiple scoring functions showed their binding potential with Sortase A. The results are proved to be a simple yet efficient choice of structure-based virtual screening. The identified compounds are druglike and show high rankings among all set protocols of the virtual screening. We hope that the study would eventually help to expedite the discovery of novel drug candidates against MRSA.

Identification and characterization of potential drug targets by subtractive genome analyses of methicillin resistant Staphylococcus aureus.

Methicillin resistant Staphylococcus aureus (MRSA) causes serious infections in humans and becomes resistant to a number of antibiotics. Due to the emergence of antibiotic resistance strains, there is an essential need to develop novel drug targets to address the challenge of multidrug-resistant bacteria. In current study, the idea was to utilize the available genome or proteome in a subtractive genome analyses protocol to identify drug targets within two of the MRSA types, i.e., MRSA ST398 and MRSA 252. Recently, the use of subtractive genomic approaches helped in the identification and characterization of novel drug targets of a number of pathogens. Our protocol involved a similarity search between pathogen and host, essentiality study using the database of essential genes, metabolic functional association study using Kyoto Encyclopedia of Genes and Genomes database (KEGG), cellular membrane localization analysis and Drug Bank database. Functional family characterizations of the identified non homologous hypothetical essential proteins were done by SVMProt server. Druggability potential of each of the identified drug targets was also evaluated by Drug Bank database. Moreover, metabolic pathway analysis of the identified druggable essential proteins with KEGG revealed that the identified proteins are participating in unique and essential metabolic pathways amongst MRSA strains. In short, the complete proteome analyses by the use of advanced computational tools, databases and servers resulted in identification and characterization of few nonhomologous/hypothetical and essential proteins which are not homologous to the host genome. Therefore, these non-homologous essential targets ensure the survival of the pathogen and hence can be targeted for drug discovery.

Remifentanil infusion prolongs spinal anesthesia.

Spinal anesthesia was given to a patient undergoing transurethral resection ofprostate (TURP). A total of 3.2 ml of bupivacaine 0.5% mixed with fentanyl 20 mcg were used. The patient started experiencing sensation after 150 min. Remifentanil intravenous infusion prolonged the duration of anesthesia for an additional 105 minutes.

Anaesthetic management of a patient of Brugada syndrome for an emergency appendicectomy.

Brugada syndrome is a myocardial transmembrane conduction of sodium abnormality and a common cause of sudden cardiac death. It is characterized by a distinctive electrocardiograph pattern with right bundle branch block and ST segment elevation in precordial leads V1-V3. Many factors during general anesthetic management could precipitate malignant dysrhythmia. We report the anesthetic management of a patient with Brugada syndrome for emergency appendectomy uneventfully.