Alpha thalassemia deletions found in suspected cases of beta thalassemia major in Pakistani population

Background and Objective: Alpha [alpha] thalassemia is a hereditary disorder and is caused by deletions or mutations in globin genes. It is present in two clinically significant forms: hemoglobin Bart hydrops fetalis [Hb Bart] syndrome and hemoglobin H [HbH] disease. It is highly prevalent in South-East Asia or Mediterranean countries. The most common deletion reported in alpha thalassemia in Pakistani population was -alpha[3.7] with a frequency of 8.3%, and the rare forms were -alpha[4.2] [0.2%] and alphaalphaalpha[anti3.7] [0.9%]. In our study, diagnosis of severe anemia cases without any alpha and beta mutations or deletions were made by using extended alpha thalassemia deletions panel. The main objective of this study was to determine the prevalence and to study the spectra of alpha thalassemia gene deletions in beta thalassemia patients with the use of an extended panel including -[SEA], --[FIL], --[MED], --[20.5], --[THAI] in addition to -alpha [3.7], -alpha [4.2] and - alphaalphaalpha[anti3.7]

Methods: The samples were collected in ethylenediaminetetraacetic acid [EDTA] vacutainers. A total of 156 samples were analyzed for alpha thalassemia mutations. This cohort included 121 samples of beta thalassemia major, nine samples of beta thalassemia minor and 26 without any evidence of beta thalassemia mutations. DNA was extracted with Qiagen extraction kit. The primers for determination of different subsets of alpha thalassemia deletions were included. PCR amplification was performed and result interpreted on agarose gel

Results: Co-inheritance of alpha thalassemia [-alpha [3.7], -alpha [4.2]] with homozygous beta thalassemia was detected in 30% cases of studied cohort [37 out of 121]. The most common found was -alpha[3.7] deletion [35/37] as single/double deletions or in combination with -alphaalphaalpha[anti3.7]. In undiagnosed cases screened for beta thalassemia major, we found Mediterranean [-alpha [MED]] deletion at specifically 875 bp on agarose gel. This is distinctive finding in case of detecting -alpha [MED] instead of any other deletion from Pakistan

Conclusion: Alpha thalassemia deletions [-alpha[3.7], -alpha[4.2]] are the common co-inherited deletions found in beta thalassemia major patients. On the basis of results, we propose an extended alpha thalassemia genetic mutation panel should be used for screening of children presenting with anemia with suspicion of haemoglobinopathy

Homozygous mutations in NTRK1 gene underlie congenital insensitivity to pain with anhidrosis in Pakistani families

Congenital insensitivity to pain with anhidrosis is a rare autosomal recessive disorder presenting with loss of pain sensation, thermal sensation defects, and self-mutilating behavior. In the present study, we recruited two consanguineous pedigree showing pain insensitivity symptoms from Pakistan for clinical and molecular investigations. In family A, one female patient displayed classical CIPA symptoms along with microcephaly and severe intellectual disability. During course of the disease, her right foot was amputated and had remarkable dental degeneration and teeth shedding. In family B, one boy presented with classical symptoms of congenital insensitivity to pain with anhidrosis. Blood was collected from both families for molecular studies. Sequencing with the Ilumina Trusight One Sequencing Panel covering 4813 OMIM genes revealed a known homozygous mutation c.2084C>T; p.P695L of NTRK1 in family A and a novel truncated mutation c.2025C>G; p.Y681X in family B. Protein modeling analysis of both mutations (p.P695L and p.Y681X) predicted loss of the rigidity in tyrosine kinase domain of NTRK1 that led to conformational changes as well as deleterious effect on protein function. The known mutation was reported more than a decade ago in a family from Northern Israel and other non-sense mutation is newly identified. It is interested that most of NTRK1 mutations are associated with this domain. This is first ever report of NTRK1 variants in congenital insensitivity to pain with anhidrosis patients from Pakistan.

Timing of Levothyroxine in the Treatment of Primary Hypothyroidism.

Aim: Timing of levothyroxine (L-thyroxine) administration seems beneficial for early obtaining thyroid state. The present study aimed at investigating the best time of L-thyroxine administration that can achieve earlier normalization of thyroid stimulating hormone (TSH) and free thyroxine (FT4) levels in patients with primary hypothyroidism. Study Design: Eighty two patients with primary hypothyroidism were recruited between November 2012 and July 2013 during their consultation to Al-Faiha Specialized Diabetes, Endocrine and Metabolism Center, Basrah, Iraq. The patients were divided into two equal groups; group A were receiving L-thyroxine daily, one hour before breakfast, group B: the dose of L-thyroxine was given at the evening. TSH, FT4, Body mass index (BMI), blood pressure, lipid profile were measured before, 30, 60 and 90 days after treatment with L-thyroxine. Results: The mean reduction in TSH from baseline for the evening treatment was 13.6±22.2 mIU/ml which was slightly and insignificantly higher than the value of the morning treatment (11.3±22.5 mIU/ml), P = .63, df = 80, 95% CI: -12.17, 7.5). The mean increase in FT4 from baseline for the evening treatment was 5.7±4.9 pmol/l which was lower than 7.6±6 pmol/l in the morning treatment, (P = .12, df = 80, 95% CI: - 0.5, 4.3). There was no effect of treatment timing on lipid profile, blood pressure, and BMI. Conclusions: There were no differences between the morning and evening treatment with L-thyroxine on early normalization of TSH and FT4.

A significant hepatotoxicity associated with paracetamol overdose in the absence of kidney injury in rabbits.

Background: Overdose of paracetamol (PCM) is reported to cause hepatotoxicity and nephrotoxicity, or nephrotoxicity in absence of hepatotoxicity. This study was planned to investigate hepatotoxicity or nephrotoxicity induced by PCM. Methods: Two groups of rabbits, six rabbits in each were used; control group were treated with normal saline, the second group was treated with PCM 1 g/kg/day orally for 9 days. Results: PCM lead to a significant rise in serum liver enzymes, aspartate aminotransferase, alanine transaminase, alkaline phosphatase and total bilirubin with an increase in serum level of malondialdehyde (MDA) and reduction in serum glutathione (GSH). MDA level in liver homogenate was also significantly increased. These findings were further confirmed by histopathological changes suggestive of severe liver damage. On the contrary, PCM slightly increased serum creatinine, without changing MDA and GSH in kidney homogenate. Lack of PCM nephrotoxicity was further confirmed by histopathological examination. Conclusion: PCM overdose produced severe hepatotoxicity without affecting the kidneys of the rabbits.