Pooling strategy to construct in-house high-resolution electrospray ionization tandem mass spectrometry database of drugs

Adeeba Khadim,Bibi Zareena,Saddam Hussain,Syed Usama Yaseen Jeelani,Arslan Ali,Syed Ghulam Musharraf 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.107 No.-

High-throughput and efficient identification of drugs in a complex mixture is required in many pharmaceuticalindustries and drug testing laboratories. The present study aims to utilize an efficient poolingstrategy for the preparation of high-resolution tandem mass spectrometry (HR-MS/MS) library of drugswith different therapeutic properties. Compounds were classified into four classes of low, medium–low,medium–high and high log P values and were pooled into five pools (100 drugs pool). Compounds withisobaric and close log p values were placed in separate groups to overcome the co-elution problem. Liquidchromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) was utilized to generate ahigh-resolution tandem mass spectrometry (MS/MS) spectral library in both positive and negative electrosprayionization modes of 491 drugs. Spectra were collected by LC-MS/MS analysis using system automatedcollision energy i.e. of 25–60 eV and four predetermined collision energies (10, 20, 30 and 40 eV)for each compound using schedule precursor list of [M + H]+ and [M + Na]+ ions. To validate the applicabilityof the library, serum samples spiked at three concentrations close to bioavailability (0.308 ng/mL,3.08 ng/mL and 30.08 ng/mL) of drugs and were analyzed by LC-QTOF-MS/MS. The HRMS library searchsuccessfully identified all compounds present in the spiked serum samples based on exact masses of precursorions, MS/MS data, and retention time etc. The accurate mass LC-QTOF-MS based tandem massspectral libraries represent a useful tool for the identification of drugs in clinical samples, for pharmacologicaland forensic screening applications etc.

Conducting polymer-based electrochemical biosensors for neurotransmitters: A review

Moon, Jong-Min,Thapliyal, Neeta,Hussain, Khalil Khadim,Goyal, Rajendra N.,Shim, Yoon-Bo Elsevier 2018 Biosensors & bioelectronics Vol.102 No.-

<P><B>Abstract</B></P> <P>Neurotransmitters are important biochemical molecules that control behavioral and physiological functions in central and peripheral nervous system. Therefore, the analysis of neurotransmitters in biological samples has a great clinical and pharmaceutical importance. To date, various methods have been developed for their assay. Of the various methods, the electrochemical sensors demonstrated the potential of being robust, selective, sensitive, and real time measurements. Recently, conducting polymers (CPs) and their composites have been widely employed in the fabrication of various electrochemical sensors for the determination of neurotransmitters. Hence, this review presents a brief introduction to the electrochemical biosensors, with the detailed discussion on recent trends in the development and applications of electrochemical neurotransmitter sensors based on CPs and their composites. The review covers the sensing principle of prime neurotransmitters, including glutamate, aspartate, tyrosine, epinephrine, norepinephrine, dopamine, serotonin, histamine, choline, acetylcholine, nitrogen monoxide, and hydrogen sulfide. In addition, the combination with other analytical techniques was also highlighted. Detection challenges and future prospective of the neurotransmitter sensors were discussed for the development of biomedical and healthcare applications.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Introduction to electrochemical biosensors for neurotransmitters. </LI> <LI> Comprehensive overview of conducting polymers for biosensors. </LI> <LI> Conducting polymer-based electrochemical neurotransmitters sensors. </LI> <LI> Future trends in point-of-care analysis. </LI> </UL> </P>

Molecular and in silico analyses validates pathogenicity of homozygous mutations in the <i>NPR2</i> gene underlying variable phenotypes of <i>Acromesomelic dysplasia</i>, type Maroteaux

Irfanullah,Zeb, Amir,Shinwari, Naila,Shah, Khadim,Gilani, Syed Zohaib Tayyab,Khan, Saadullah,Lee, Keun Woo,Raza, Syed Irfan,Hussain, Shabir,Liaqat, Khurram,Ahmad, Wasim Pergamon 2018 The international journal of biochemistry & cell b Vol.102 No.-

<P><B>Abstract</B></P> <P>Homozygous and/or heterozygous loss of function mutations in the natriuretic peptide receptor B (<I>NPR2</I>) have been reported in causing acromesomelic dysplasia, type Maroteaux with variable clinical features and idiopathic short stature with nonspecific skeletal deformities. On the other hand, gain of function mutations in the same gene result in overgrowth disorder suggesting that NPR2 and its ligand, natriuretic peptide precursor C (CNP), are the key players of endochondral bone growth. However, the precise mechanism behind phenotypic variability of the <I>NPR2</I> mutations is not fully understood so far.</P> <P>In the present study, three consanguineous families of Pakistani origin (A, B, C) with variable phenotypes of acromesomelic dysplasia, type Maroteaux were evaluated at clinical and molecular levels. Linkage analysis followed by Sanger sequencing of the <I>NPR2</I> gene revealed three homozygous mutations including p.(Leu314 Arg), p.(Arg371*), and p.(Arg1032*) in family A, B and C, respectively. In silico structural and functional analyses substantiated that a novel missense mutation [p.(Leu314 Arg)] in family A allosterically affects binding of NPR2 homodimer to its ligand (CNP) which ultimately results in defective guanylate cyclase activity. A nonsense mutation [p.(Arg371*)] in family B entirely removed the transmembrane domain, protein kinase domain and guanylate cyclase domains of the NPR2 resulting in abolishing its guanylate cyclase activity. Another novel mutation [p.(Arg1032*)], found in family C, deteriorated the guanylate cyclase domain of the protein and probably plundered its guanylate cyclase activity. These results suggest that guanylate cyclase activity is the most critical function of the NPR2 and phenotypic severity of the NPR2 mutations is proportional to the reduction in its guanylate cyclase activity.</P>

Diagnostic Accuracy, Sensitivity, Specificity and Positive Predictive Value of Fine Needle Aspiration Cytology (FNAC) in Intra Oral Tumors

Gillani, Munazza,Akhtar, Farhan,Ali, Zafar,Naz, Irum,Atique, Muhammad,Khadim, Muhammad Tahir Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.8

Objective: The objective of this study was to establish the diagnostic accuracy, specificity and sensitivity of fine needle aspiration cytology(FNAC) for intra-oral tumors, comparing with histopathology as the gold standard. Materials and methods: Forty cases of FNA cytology from intraoral tumors was performed in AFID along with the demographic data and clinical information and then diagnosed at AFIP, Rawalpindi. Then the cytology results obtained per FNAC were compared with the histopathological biopsy results of the same lesions. The following variables were recorded for each patient: Age, gender, site of biopsy, diagnosis. The data were entered and analyzed using Open-epi version 2.0. Diagnostic accuracy, sensitivity, specificity, positive predictive value and negative predictive value were calculated. Cohen Kappa was further applied to compare the agreement between the biopsy and FNAC diagnoses. A p-value of < 0.05 was considered as statistically significant. Results: Among the total patients included in the study there were 24 males and 16 females, with a ratio of 1.5:1. Age of the patients ranged from 24 to 80 years with a mean of 52 years. A total of six sites were aspirated from the oral cavity with maximum (11) aspirates taken from alveolar ridge. The results of FNAC revealed that there were 32 malignant and 8 benign aspirates. Confirmation through histopathological analysis came for 31/32 malignant cases while one was falsely given positive for malignancy on FNAC. Among a total of 40 cases, 31(77%) cases diagnosed were found to be malignant and remaining 9(23%) were benign. The FNAC results revealed 32 malignant and 8 benign lesions. Histopathology of the subsequent surgically excised specimen showed malignant lesions in 31(77%) and benign in 9(23%) patients. As a whole, it was found that the absolute sensitivity for introral FNAC was 100% and specificity 89% with positive predictive value of 97% and negative predictive value of 100%. Conclusion: Cytological diagnosis was almost corroborative with final histopathological diagnosis in all cases, with very few exceptions, exhibiting high diagnostic accuracy.

Efficient power allocation algorithm in downlink cognitive radio networks

Omar Abdulghafoor,Musbah Shaat,Ibraheem Shayea,Farhad E. Mahmood,Rosdiadee Nordin,Ali Khadim Lwas 한국전자통신연구원 2022 ETRI Journal Vol.44 No.3

In cognitive radio networks (CRNs), the computational complexity of resource allocation algorithms is a significant problem that must be addressed. However, the high computational complexity of the optimal solution for tackling resource allocation in CRNs makes it inappropriate for use in practical applications. Therefore, this study proposes a power-based pricing algorithm (PPA) primarily to reduce the computational complexity in downlink CRN scenarios while restricting the interference to primary users to permissible levels. A twostage approach reduces the computational complexity of the proposed mathematical model. Stage 1 assigns subcarriers to the CRN’s users, while the utility function in Stage 2 incorporates a pricing method to provide a power algorithm with enhanced reliability. The PPA’s performance is simulated and tested for orthogonal frequency-division multiplexing-based CRNs. The results confirm that the proposed algorithm’s performance is close to that of the optimal algorithm, albeit with lower computational complexity of O(Mlog(M)).

Reproductive fecundity of Iraqi Awassi ewes immunized against synthetic inhibin-α subunit or steroid-free bovine follicular fluid

Al-Sa'aidi, Jabbar Abbas Ahmed,Khudair, Khalisa Khadim,Khafaji, Sura Safi Asian Australasian Association of Animal Productio 2018 Animal Bioscience Vol.31 No.8

Objective: The present study was conducted to investigate the impacts of active and passive immunization against synthetic inhibin and steroid-free bovine follicular fluid, respectively, on reproductive fecundity out of breeding season in Iraqi Awassi ewes. Methods: Follicular fluid was aspired from mature bovine follicles, treated with activated charcoal, and used for immunization of male rabbits for obtaining steroid free bovine follicular fluid (SFBFF) antiserum. Forty non-pregnant Awassi ewes were allocated into 4 groups (n = 10 each). At day 38 of experiment, ewes were treated with intra-vaginal MPA sponge (60 mg for 12 days). At days 0, 28, and 50, ewes were treated with 4, 2, and 2 mL of normal saline (control; C-ve), 400, 200, and $200{\mu}g$ of ovalbumin (C+ve), 400, 200 and $200{\mu}g$ of inhibin (SI group), respectively, and 4 mL of normal saline at day 0, and 4 and 2 mL of SFBFF antiserum at days 28 and 50, respectively, (AI group). After mating with Awassi rams, pregnancy and embryo number were diagnosed, at day 38 of pregnancy, using ultrasonography. Blood samples were collected at days 30, 60, 90, and 120 of pregnancy, for assessment of estradiol-$17{\beta}$ (E2) and progesterone (P4) levels. After parturition, numbers of delivered lambs were recorded. Results: The results revealed significant increase of P4 and significant decrease of E2 levels in SI and AI pregnant ewes than controls at days 30, 60, 90, and 120. Newborn number increased significantly in SI and AI treated than control ewes. Conclusion: Active or passive immunization against endogenous inhibin could augment reproductive fecundity out of breeding season in Iraqi Awassi ewes.

Clinicohematological parameters and outcomes in a cohort of chronic lymphocytic leukemia patients with Deletion 17p from Pakistan

Rafia Mahmood,Saleem Ahmed Khan,Chaudhry Altaf,Hamid Saeed Malik,Muhammad Tahir Khadim 대한혈액학회 2018 Blood Research Vol.53 No.4

BackgroundChronic lymphocytic leukemia (CLL) exhibits profound heterogeneity in its clinical course. Its clinicohematological and cytogenetic features play a significant role in de-termining the clinical course and in predicting the treatment response and prognosis. In this context, 17p deletion is known to predict a poor prognosis, as these cases are re-fractory to conventional therapy. This study aimed to evaluate the clinicohematological characteristics, outcomes, and prognostic factors among CLL patients with and without del 17p in Pakistan.MethodsThis prospective observational study was conducted at the Department of Haematology, Armed Forces Institute of Pathology (Rawalpindi, Pakistan) between January 2013 and December 2017. Patients were diagnosed based on the International Workshop on Chronic Lymphocytic Leukaemia IWCLL criteria, their clinicohematological parameters were recorded, and cytogenetic analyses were performed. The time from diagnosis to treatment and the 2-year overall survival rate were also evaluated.ResultsWe evaluated 130 CLL cases, including 24 patients (18.5%) with del 17p, who included 18 men (75%) and 6 women (25%). The median age was 68 years. Binet stage C was de-tected at the presentation in 16 patients (67%). Treatment was administered to 14 patients (70%) at a median interval of 11 months (range, 0‒28 mo) after diagnosis. The overall response rate was 64.3%, the median event-free survival was 9 months (range, 1‒23 mo), and the 2-year overall survival rate was 65%.ConclusionDel 17p is relatively common in Pakistan, and patients harboring this deletion had poor treatment response and survival outcomes.