Acute and sub-acute toxicity study of a Pakistani polyherbal formulation

Abstract Background Herbology is the prevailing system among the nationally-accepted alternative or complementary systems of medicine. The system is due to its general and patient-oriented methodology, is widely used in the general population exposing them to the risk of the side effects of the herbal medicines. Method The aim of study was to assess the acute and sub-acute toxicity of the polyherbal formulation Hab-e-Kabad Noshadri tablets. In the acute arm of the study, a single dose of 2000 mg/kg was administered to Swiss Albino mice which were observed for physical symptoms and behavioral changes for 72 h. In sub-acute toxicity study repeated doses of the polyherbal preparation was administered to Wistar rats of both genders, separately. The animals received three doses of polyherbal product (50 mg/kg/day, 100 mg/kg/day and 200 mg/kg/day) for a period of 28 days. On 28th day of experiment, blood sampling of animals was done for hematological and biochemical analysis i.e. liver and renal function parameters, lipid profile and then sacrificed for histopathological examination of liver and kidney. Result There was no morbidity and mortality noticed with single dose administration in acute toxicity study in mice. In sub-acute toxicity study, morphological changes with some damage in liver and kidney tissues of male and female animals were recorded at dose of 100 mg/kg/day and 200 mg/kg/day. Conclusions It was found that prolonged use at higher dose i.e. 200 mg/kg/day of this polyherbal formulation should be avoided and practitioners should cautiously prescribe this formulation in patients with hepatic and renal impairment

Molecular Diagnosis of Fragile X Syndrome in Subjects with Intellectual Disability of Unknown Origin: Implications of Its Prevalence in Regional Pakistan

<div><p>Fragile-X syndrome (FXS) is the most common form of inherited intellectual disability (ID) and affects 0.7–3.0% of intellectually compromised population of unknown etiology worldwide. It is mostly caused by repeat expansion mutations in the <i>FMR1</i> at chromosome Xq27.3. The present study aimed to develop molecular diagnostic tools for a better detection of FXS, to assess implementation of diagnostic protocols in a developing country and to estimate the prevalence of FXS in a cohort of intellectually disabled subjects from Pakistan. From a large pool of individuals with below normal IQ range, 395 subjects with intellectual disability of unknown etiology belonging to different regions of the country were recruited. Conventional-PCR, modified-PCR and Southern blot analysis methods were employed for the detection of CGG repeat polymorphisms in the <i>FMR1</i> gene. Initial screening with conventional-PCR identified 13 suspected patients. Subsequent investigations through modified PCR and Southern blot analyses confirmed the presence of the <i>FMR1</i> mutation, suggesting a prevalence of 3.5% and 2.8% (mean 3.3%) among the male and female ID patients, respectively. These diagnostic methods were further customized with the in-house conditions to offer robust screening of referral patients/families for diagnostics and genetic counseling. Prescreening and early diagnosis are crucial for designing a prudent strategy for the management of subjects with ID. Outcome of the study recommends health practitioners for implementation of molecular based FXS diagnosis in routine clinical practice to give a better care for patients similar to the ones included in the study.</p></div

A. Primer pair 1 PCR amplification products of FXS negative subjects (lanes 1–5).

<p>Agarose gel electrophoresis results of methylation specific PCR. <b>B.</b> Primer pair 3 gave amplification product of ~80bp of FXS positive subjects (lanes 2–4). <b>C.</b> Primer pair 4 did not give amplification of the FXS positive subjects (lanes 2&3) but yielded a product of ~300bp with FXS negative subjects (lane 4).</p

Survey plan for screening of subjects.

<p>Survey plan for screening of subjects.</p

Clinical features of subjects with ID of unknown origin (n = 395, confirmed FXS = 13).

<p>* metacarpophalangeal.</p><p>Clinical features of subjects with ID of unknown origin (n = 395, confirmed FXS = 13).</p

Facial features of unrelated Fragile X patients.

<p>Facial features of unrelated Fragile X patients.</p

Selected pedigrees of FXS patients.

<p>Analyzed patients and carrier mothers are given identification numbers.</p

Southern blot analysis shows a normal female control (lane 2); premutation carrier females (lanes 3 and 7); methylation mosaic male (lane 4); full mutation female (lane 5); size mosaic males (lanes 6 and 8).

<p>1kb DNA size marker is shown in lane 1.</p