Ayurveda therapy in the management of epilepsy.

Epilepsy, a chronic non-communicable disease of the brain, is one of the most common neurological diseases globally that affects people of all ages. The existence of medical, neurological, psychiatric, and cognitive comorbidities has always undermined the available advanced treatment strategies for epilepsy. New-generation antiepileptic drugs being less successful in completely controlling the seizures and observance of complex diseases, including drug-resistant cases, have provided scope for integrating and incorporating the therapeutic modalities of Ayurveda, the ancient Indian art of holistic medicine, in the effective management of epilepsy. Epilepsy can be correlated to Apasmara, described in the classics of Ayurveda as the transient appearance of unconsciousness with loathsome expression due to derangement of memory, intelligence, and mind. The multifaceted therapeutic approach of Ayurveda, which involves pharmacologic and nonpharmacologic measures, purificatory and pacifying procedures, herbal and herbo-mineral formulations, disease, and host-specific approaches, have enhanced the potential of not only relieving symptoms but also modifying the pathophysiology of the disease. Newer paradigms of research in Ayurveda, along with holistic and integrative approaches with contemporary medicine, can not only benefit the existing healthcare system but also impact future healthcare management in epileptology research. This cursory literature review is an earnest attempt to identify, evaluate, and summarize various studies and provide a comprehensive insight into the potential of Ayurveda in understanding and treating epilepsy.

Bilateral variation in the branching pattern of the subclavian artery: an unusual finding with clinical implications.

The subclavian artery is a significant branch of the aortic arch. We present a rare case of a bilateral variation in the branching pattern of the subclavian artery, observed in an adult male cadaver aged 70 years. On both the sides of the neck, all the branches of the subclavian artery took their origin from its first part. There was a rare occurrence of a cervicodorsoscapular trunk, which gave rise to superficial cervical, suprascapular, and dorsal scapular arteries. The same branching pattern was observed on the left side of the neck, with the presence of another cervicodorsoscapular trunk. Thyrocervical trunk and transverse cervical artery were both absent from the cervical region bilaterally. The inferior thyroid artery was a direct branch from the subclavian artery. Knowledge regarding variations of the subclavian artery is very important as lateral cervical region arteries are important for flap harvesting in plastic and reconstruction surgery. Preoperative radiologic evaluation of pedicles might help in choosing the optimal flap design, prevent ischemic complications, and help to improve overall treatment outcomes.

A artéria subclávia é um ramo significativo do arco da aorta. Apresentamos um caso raro de variação bilateral do padrão de ramificação da artéria subclávia, observada em um cadáver adulto do sexo masculino de 70 anos. Em ambos os lados do pescoço, todos os ramos da artéria subclávia originavam-se de sua primeira parte. Houve rara ocorrência de tronco escapular cervical dorsal, que deu origem às artérias cervical superficial, supraescapular e escapular dorsal. O mesmo padrão de ramificação foi observado no lado esquerdo do pescoço, com a presença de tronco escapular cervical dorsal. O tronco tireocervical e a artéria cervical transversa estavam ausentes em ambas as regiões cervicais direita e esquerda. A artéria tireóidea inferior consistia em um ramo direto da artéria subclávia. O conhecimento das variações da artéria subclávia é fundamental, pois as artérias da região cervical lateral são importantes para a obtenção de retalhos em cirurgias plásticas e reconstrutivas. A avaliação radiológica pré-operatória dos pedículos pode ajudar na escolha do desenho ideal do retalho, prevenir complicações isquêmicas e ajudar a melhorar o resultado geral do tratamento.

Rare variations in the Origin, Branching Pattern and Course of the Celiac Trunk: Report of Two Cases.

Multiple anomalies in the celiac arterial system presents as rare vascular malformations, depicting deviations of the normal vascular developmental pattern. We found a common left gastro-phrenic trunk and a hepato-spleno-mesenteric trunk arising separately from the abdominal aorta in one cadaver. We also found a common hepatic artery and a gastro-splenic trunk arising individually from the abdominal aorta in another cadaver. Even though many variations in the celiac trunk have been described earlier, the complex variations described here are not mentioned and classified by earlier literature. Knowledge of such variations has significance in the surgical and invasive arterial radiological procedures in the upper abdomen.

Cell and molecular targeted therapies for diabetic retinopathy.

Diabetic retinopathy (DR) stands as a prevalent complication in the eye resulting from diabetes mellitus, predominantly associated with high blood sugar levels and hypertension as individuals age. DR is a severe microvascular complication of both type I and type II diabetes mellitus and the leading cause of vision impairment. The critical approach to combatting and halting the advancement of DR lies in effectively managing blood glucose and blood pressure levels in diabetic patients; however, this is seldom achieved. Both human and animal studies have revealed the intricate nature of this condition involving various cell types and molecules. Aside from photocoagulation, the sole therapy targeting VEGF molecules in the retina to prevent abnormal blood vessel growth is intravitreal anti-VEGF therapy. However, a substantial portion of cases, approximately 30-40%, do not respond to this treatment. This review explores distinctive pathophysiological phenomena of DR and identifiable cell types and molecules that could be targeted to mitigate the chronic changes occurring in the retina due to diabetes mellitus. Addressing the significant research gap in this domain is imperative to broaden the treatment options available for managing DR effectively.

A rare congenital anomaly of the pancreas: a cadaveric case report.

CONTEXT: The pancreas is formed by ventral and dorsal pancreatic buds which arise from the endodermal lining of the gut. When the duodenum rotates to the right, the ventral pancreatic bud migrates dorsally and finally come and lie below the dorsal pancreatic bud. The developmental errors in the rotation of these components lead to annular pancreas. CASE REPORT: We report a rare type of the incomplete annular pancreas around the contents of right free margin of lesser omentum. There was an extra pancreatic mass situated horizontally immediately above the superior border of the pancreas, situated behind the lesser omentum. The right end of this mass extended into the epiploic foramen and incompletely encircled the portal vein, bile duct and hepatic artery proper from behind. The left end of the extra pancreatic mass was extended towards the lesser curvature of the stomach. Further, this mass completely surrounded the origin of three branches of the celiac trunk. Its right end was found to be continuous with the head of the pancreas, close to the pylorus. Histology of the extra pancreatic mass revealed the presence of normal pancreatic tissue. CONCLUSION: preoperative diagnosis of this rare anomaly is of clinical importance during surgeries involving the contents of right free margin of lesser omentum and epiploic foramen.

Trigonella foenum-graecum L. seed extract modulates biochemical and histomorphological changes in therapeutic model of high-fat diet-fed ovariectomized rats.

This study investigates the therapeutic effect of petroleum ether fraction of Trigonella foenum-graecum L. (PE-TFG) seed extract in ovariectomized rats fed with high-fat diet. Rats were randomly grouped into sham ovariectomy (S.OVX), ovariectomy + high-fat diet (OVX + HFD), and treatment groups. The blood samples were collected, and lipid profile, glucose, hepatic markers, and inflammatory markers were estimated. Liver, kidney, and common carotid artery were isolated for histopathological observations. Liver samples were tested for antioxidant, oxidative stress markers, mRNA expression of adiponectin, and PPAR-γ. PE-TFG treatment significantly decreased total cholesterol (18%), LDL (20%), hepatic markers (28%), leptin (17%), TNF-α (21%), and increased mRNA expression of adiponectin and PPAR-γ. There was also micro- and macro-hepatic steatosis, inflammation in the liver, deteriorated tubules in the kidney, and increased tunica intima and media thickness of the common carotid artery. These pathological alterations were reversed with PE-TFG administration. This impact might be linked to phytoestrogens and other components in PE-TFG such as diosgenin, phenols, and flavonoids. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03707-8.

Oncogenic BRAFV600E induces expression of neuronal differentiation marker MAP2 in melanoma cells by promoter demethylation and down-regulation of transcription repressor HES1.

MAP2 is a neuron-specific microtubule-associated protein that binds and stabilizes dendritic microtubules. Previously, we showed that MAP2 expression is (a) activated in cutaneous primary melanoma and (b) inversely associated with melanoma tumor progression. We also showed that ectopic expression of MAP2 in metastatic melanoma cells inhibits cell growth by inducing mitotic spindle defects and apoptosis. However, molecular mechanisms of regulation of MAP2 gene expression in melanoma are not understood. Here, we show that in melanoma cells MAP2 expression is induced by the demethylating agent 5-aza-2'-cytidine, and MAP2 promoter is progressively methylated during melanoma progression, indicating that epigenetic mechanisms are involved in silencing of MAP2 in melanoma. In support of this, methylation of MAP2 promoter DNA in vitro inhibits its activity. Because MAP2 promoter activity levels in melanoma cell lines also correlated with activating mutation in BRAF, a gene that is highly expressed in neurons, we hypothesized that BRAF signaling is involved in MAP2 expression. We show that hyperactivation of BRAF-MEK signaling activates MAP2 expression in melanoma cells by two independent mechanisms, promoter demethylation or down-regulation of neuronal transcription repressor HES1. Our data suggest that BRAF oncogene levels can regulate melanoma neuronal differentiation and tumor progression.

Modulation of inflammatory markers by petroleum ether fraction of Trigonella foenum-graecum L. seed extract in ovariectomized rats.

This study evaluates the modulation of inflammatory markers by petroleum ether fraction of Trigonella foenum-graecum L. (PE-TFG) seed extract in ovariectomized rats. The HPTLC method was used for standardization and to quantify the diosgenin in PE-TFG. For testing PE-TFG in rats, the total duration of treatment was 12-weeks, and the rats were sacrificed on week 12. The tissue samples such as blood, liver, heart, and aorta were isolated for testing inflammatory markers such as adiponectin, leptin, PPAR-γ, TNF-α, lipid profile, hepatic markers, antioxidants, and oxidative stress markers. The PE-TFG treatment decreased the elevation of total cholesterol, triglyceride, AST, and ALT. Upon PE-TFG treatment, there was a significant increase in adiponectin and PPAR-γ mRNA expression. Leptin and TNF-α were normal after treatment with PE-TFG seed extract. Further, micro-steatosis of hepatocytes marked glomerular hypertrophy in the kidney and increased thickness of tunica intima and media of common carotid artery was reversed after treatment with PE-TFG. PRACTICAL APPLICATIONS: Trigonella foenum-graecum L. is a curative plant used to treat inflammatory conditions like diabetes, obesity, dyslipidemia, arthritis, cancer, and digestive disorders. In our study, PE-TFG supplementation has a protective effect on OVX-induced inflammation, oxidative stress, mRNA expression of adiponectin and PPAR-γ, hepatic steatosis, and decreased thickness of tunica intima and media of common carotid artery.

Supplementation of fenugreek with choline-docosahexaenoic acid attenuates menopause induced memory loss, BDNF and dendritic arborization in ovariectomized rats.

Cognitive impairment due to natural or surgical menopause is always associated with estrogen deficiency leading to reduced brain-derived neurotrophic factor (BDNF). Reduced BDNF levels in menopause affect neuronal maturation, survival, axonal and dendritic arborization and the maintenance of dendritic spine density. Conventional long-term estrogen replacement therapy reported causing the risk of venous thromboembolism and breast cancer. To overcome these undesirable effects, phytoestrogens have been used in menopause-induced condition without the risk of side effects. Therefore, the aim of the present study was to investigate the effect of dietary supplementation of fenugreek seed extract (FG) either alone or in combination with choline-DHA on BDNF and dendritic arborization of pyramidal neurons in CA1 and CA3 regions of the hippocampus in ovariectomized rats. Female Wistar rats of 9-10 months old were divided into six groups as normal control (NC); ovariectomy (OVX); OVX + FG; OVX + choline-DHA; OVX + FG + choline-DHA; and OVX + estradiol. All the groups, except NC, were ovariectomized. After 2 weeks of ovariectomy, dietary supplementation was initiated for a period of 30 days. After supplementation, behavioral studies, BDNF levels and dendritic arborization were estimated. Ovariectomized (OVX) rats showed reduced BDNF levels, dendritic branching points and dendritic intersections of pyramidal neurons in CA1 and CA3 regions of the hippocampus. OVX rats supplemented with FG with choline-DHA showed significantly improved BDNF levels, dendritic branching points and dendritic intersections. These results are demonstrating that FG with choline-DHA supplementation can be an alternative for estrogen replacement therapy to modulate menopause-induced learning and memory deficits.

Calcium homeostasis in human melanocytes: role of transient receptor potential melastatin 1 (TRPM1) and its regulation by ultraviolet light.

Transient receptor potential melastatin (TRPM) is a subfamily of ion channels that are involved in sensing taste, ambient temperature, low pH, osmolarity, and chemical ligands. Melastatin 1/TRPM1, the founding member, was originally identified as melanoma metastasis suppressor based on its expression in normal pigment cells in the skin and the eye but not in aggressive, metastasis-competent melanomas. The role of TRPM1 and its regulation in normal melanocytes and in melanoma progression is not understood. Here, we studied the relationship of TRPM1 expression to growth and differentiation of human epidermal melanocytes. TRPM1 expression and intracellular Ca(2+) levels are significantly lower in rapidly proliferating melanocytes compared to the slow growing, differentiated melanocytes. We show that lentiviral short hairpin RNA (shRNA)-mediated knockdown of TRPM1 results in reduced intracellular Ca(2+) and decreased Ca(2+) uptake suggesting a role for TRPM1 in Ca(2+) homeostasis in melanocytes. TRPM1 knockdown also resulted in a decrease in tyrosinase activity and intracellular melanin pigment. Expression of the tumor suppressor p53 by transfection or induction of endogenous p53 by ultraviolet B radiation caused repression of TRPM1 expression accompanied by decrease in mobilization of intracellular Ca(2+) and uptake of extracellular Ca(2+). These data suggest a role for TRPM1-mediated Ca(2+) homeostasis, which is also regulated by ultraviolet B, in melanogenesis.

MAGE-A, mMage-b, and MAGE-C proteins form complexes with KAP1 and suppress p53-dependent apoptosis in MAGE-positive cell lines.

The MAGE-A, MAGE-B, and MAGE-C protein families comprise the class-I MAGE/cancer testes antigens, a group of highly homologous proteins whose expression is suppressed in all normal tissues except developing sperm. Aberrant expression of class I MAGE proteins occurs in melanomas and many other malignancies, and MAGE proteins have long been recognized as tumor-specific targets; however, their functions have largely been unknown. Here, we show that suppression of class I MAGE proteins induces apoptosis in the Hs-294T, A375, and S91 MAGE-positive melanoma cell lines and that members of all three families of MAGE class I proteins form complexes with KAP1, a scaffolding protein that is known as a corepressor of p53 expression and function. In addition to inducing apoptosis, MAGE suppression decreases KAP1 complexing with p53, increases immunoreactive and acetylated p53, and activates a p53 responsive reporter gene. Suppression of class I MAGE proteins also induces apoptosis in MAGE-A-positive, p53wt/wt parental HCT 116 colon cancer cells but not in a MAGE-A-positive HCT 116 p53-/- variant, indicating that MAGE suppression of apoptosis requires p53. Finally, treatment with MAGE-specific small interfering RNA suppresses S91 melanoma growth in vivo, in syngenic DBA2 mice. Thus, class I MAGE protein expression may suppress apoptosis by suppressing p53 and may actively contribute to the development of malignancies and by promoting tumor survival. Because the expression of class I MAGE proteins is limited in normal tissues, inhibition of MAGE antigen expression or function represents a novel and specific treatment for melanoma and diverse malignancies.

Influence of traditional medicines on the activity of keratinocytes in wound healing: an in-vitro study.

Natural medicinal systems such as Ayurveda and folk medicine has remedies for wound management. However, the exact cellular and extracellular mechanisms involved in the healing process and its influence on keratinocytes is less discussed. Therefore, the present study was designed to evaluate the effect of certain natural wound healing medicines on the biology of the keratinocytes/HaCaT cells. Test materials such as honey (H), ghee (G), aqueous extracts of roots of Glycyrrhiza glabra (GG) and leaves of Nerium indicum (NI) were considered. The HaCaT cells were treated with the test materials singly and in combinations (H+G, all combined [Tot]) for a specific period (24, 48, and 72 hours). The cells were then subjected to cytotoxicity/proliferation and migration/scratch assays. All the test materials, except NI, were non-cytotoxic and showed increased cell proliferation at variable concentrations. Significant observations were made in the groups treated with honey (100 µg/ml at 48 hours, P<0.05; 1,000 µg/ml at 72 hours, P<0.05), GG (all concentrations at 48 hours, P<0.05; 750 µg/ml at 72 hours, P<0.05), H+G (250 µg/ml at 24 hours, P<0.001; 500 µg/ml at 48 and 72 hours, P<0.05), and Tot (50 µg/ml at 24, 48 and 72 hours, P<0.01). In the in-vitro wound healing assay, all the treated groups showed significant migration and narrowing of the scratch area by 24 and 48 hours (P<0.001) compared to control. The results obtained from the present study signifies the positive influence of these natural wound healing compounds on keratinocytes/HaCaT cells.

Microtubule-associated proteins as targets in cancer chemotherapy.

Natural and synthetic compounds that disrupt microtubule dynamics are among the most successful and widely used cancer chemotherapeutic agents. However, lack of reliable markers that predict sensitivity of cancers to these agents and development of resistance remain vexing issues. There is accumulating evidence that a family of cellular proteins that are associated with and alter the dynamics of microtubules can determine sensitivity of cancer cells to microtubule-targeting agents and play a role in tumor cell resistance to these agents. This growing family of microtubule-associated proteins (MAP) includes products of oncogenes, tumor suppressors, and apoptosis regulators, suggesting that alteration of microtubule dynamics may be one of the critical events in tumorigenesis and tumor progression. The objective of this review is to integrate the knowledge on these seemingly unrelated proteins that share a common function and examine their relevance to microtubule-targeting therapies and highlight MAPs-tubulin-drug interactions as a novel avenue for new drug discovery. Based on the available evidence, we propose that rational microtubule-targeting cancer therapeutic approaches should ideally include proteomic profiling of tumor MAPs before administration of microtubule-stabilizing/destabilizing agents preferentially in combination with agents that modulate the expression of relevant MAPs.

Transcriptional regulation of human MAP2 gene in melanoma: role of neuronal bHLH factors and Notch1 signaling.

Microtubule-associated protein 2 (MAP2), a neuron-specific protein, stabilizes microtubules and is critical for neurite outgrowth and dendrite development. Although MAP2 is widely used as a marker of neuronal differentiation, regulation of its transcription has not been investigated. We showed that MAP2 is frequently activated in human cutaneous melanoma. Here, we identified a 2.2 kb region that is sufficient for neuronal-specific expression in vitro and in vivo. Comparative analysis of the mouse, rat and human MAP2 promoter sequences showed the presence of a conserved bHLH factor binding sites. Electrophoretic mobility shift analysis, promoter mutagenesis and co-transfection experiments showed that NeuroD, a pro-neuronal differentiation factor, and Hairy and Enhancer of Split (HES1), a transcription repressor, are involved in the regulation of MAP2 promoter activity. Melanoma cells express both NeuroD and HES1. Chromatin immunoprecipitation showed that in metastatic melanoma cells N-box region of the MAP2 promoter is occupied by endogenous HES1. We show that the inhibition of Notch signaling, a regulator of HES1 gene expression, and/or shRNA knockdown of HES1 results in the upregulation of MAP2 promoter activity. Thus, our data suggest that Notch signaling, which is implicated in melanoma progression, and HES1 play a role in MAP2 gene regulation during melanoma progression.

A unique branching pattern of the axillary artery in a South Indian male cadaver.

Axillary artery divides into 3 parts by pectoralis minor muscle and classically each part has its own branches. There are many reports to show different variations in the branching pattern of the axillary artery. However, here we have shown an unreported unique branching pattern of axillary artery, where most of the branches of the axillary artery are arising from one common trunk from its 2nd part. Further, with relevant literature review we have also discussed their developmental and clinical importance (Fig. 1, Ref. 16). Full Text (Free, PDF) www.bmj.sk.

Anomalous origin of the lumbrical muscles: a study on South Indian cadavers.

Although variations in the attachments of the lumbrical muscles have been commonly reported, these have been seen mainly in the Caucasian population. The present study has been undertaken in South Indian population. The upper extremities of 24 South Indian (20 male and four female) cadavers were examined. Three instances of variant origins of the lumbrical muscles (two instances of the second and one of the first lumbrical) were seen. The muscles were unusually long extending to the level of the proximal border of the flexor retinaculum and in two of the cases it was taking origin from the flexor digitorum profundus tendon and its accessory belly. In one case it was taking origin from the flexor digitorum profundus and superficialis tendons. An anomalous origin of the lumbrical from muscles in the forearm has the potential to cause compression of the median nerve in the carpal tunnel.

Bilateral variation in the branching pattern of the subclavian artery: an unusual finding with clinical implications / Variação bilateral no padrão de ramificação da artéria subclávia: um achado incomum com implicações clínicas

Abstract The subclavian artery is a significant branch of the aortic arch. We present a rare case of a bilateral variation in the branching pattern of the subclavian artery, observed in an adult male cadaver aged 70 years. On both the sides of the neck, all the branches of the subclavian artery took their origin from its first part. There was a rare occurrence of a cervicodorsoscapular trunk, which gave rise to superficial cervical, suprascapular, and dorsal scapular arteries. The same branching pattern was observed on the left side of the neck, with the presence of another cervicodorsoscapular trunk. Thyrocervical trunk and transverse cervical artery were both absent from the cervical region bilaterally. The inferior thyroid artery was a direct branch from the subclavian artery. Knowledge regarding variations of the subclavian artery is very important as lateral cervical region arteries are important for flap harvesting in plastic and reconstruction surgery. Preoperative radiologic evaluation of pedicles might help in choosing the optimal flap design, prevent ischemic complications, and help to improve overall treatment outcomes.

Resumo A artéria subclávia é um ramo significativo do arco da aorta. Apresentamos um caso raro de variação bilateral do padrão de ramificação da artéria subclávia, observada em um cadáver adulto do sexo masculino de 70 anos. Em ambos os lados do pescoço, todos os ramos da artéria subclávia originavam-se de sua primeira parte. Houve rara ocorrência de tronco escapular cervical dorsal, que deu origem às artérias cervical superficial, supraescapular e escapular dorsal. O mesmo padrão de ramificação foi observado no lado esquerdo do pescoço, com a presença de tronco escapular cervical dorsal. O tronco tireocervical e a artéria cervical transversa estavam ausentes em ambas as regiões cervicais direita e esquerda. A artéria tireóidea inferior consistia em um ramo direto da artéria subclávia. O conhecimento das variações da artéria subclávia é fundamental, pois as artérias da região cervical lateral são importantes para a obtenção de retalhos em cirurgias plásticas e reconstrutivas. A avaliação radiológica pré-operatória dos pedículos pode ajudar na escolha do desenho ideal do retalho, prevenir complicações isquêmicas e ajudar a melhorar o resultado geral do tratamento.

Additional Muscle Slip of Bicipital Aponeurosis and its Anomalous Relationship with the Median Cubital Vein.

The cubital region of the arm is a common site for recording blood pressure, taking blood for analysis and administering intravenous therapy and blood transfusions. During the routine dissection of a 70-year-old male cadaver at the Kasturba Medical College, Manipal, Karnataka, India, in 2015, it was observed that the aponeurotic insertion of the biceps brachii muscle divided into two slips. The medial slip fused normally with the deep fascia of the forearm, while flexor carpi radialis muscle fibres originated from the lateral slip. There was also a single vein in the forearm, the cephalic vein, which bifurcated to form the median cubital vein and the cephalic vein proper. The median cubital vein, further reinforced by the radial vein, passed deep to the two slips of the bicipital aponeurosis and then continued as the basilic vein. During venepuncture, medical practitioners should be aware of potential cubital fossa variations which could lead to nerve entrapment syndromes.

A Comparison Between Phorbol 12 Myristate 13 Acetate and Phorbol 12, 13 Dibutyrate in Human Melanocyte Culture.

INTRODUCTION: Melanocyte culture is an integral part of the studies of skin biology and cosmetic applications. After the introduction of selective medium for the culture of human melanocyte using Phorbol 12-myristate13-acetate (PMA) in 1982, a lot of methods of culturing were tried but till date PMA is a preferred mitogen because of its cost effectiveness compared to growth factors. We have tried to preliminarily evaluate the efficacy of another phorbol ester, Phorbol 12, 13-dibutyrate (PDBu) in melanocyte culture because of its less hydrophobic nature compared to PMA. This property minimizes the trace amount of mitogen in cell culture after washing off and hence does not interfere in other biological assays. AIM: To evaluate the differences in the melanocyte survival rate, morphology and mitotic index when grown in media supplemented with PMA and PDBu. MATERIALS AND METHODS: Foreskins were collected from children undergoing circumcision. Epidermal cells were isolated from foreskin and cultured using PMA and PDBu. Melanocytes in culture were monitored for the better establishment and documented. In proliferative assay, melanocytes were treated with PMA and PDBu for 24, 48 and 72 hours and proliferation was measured using 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay method. RESULTS: When cultured, melanocytes acquired proliferative status and bipolar morphology quicker in PDBu medium than in PMA medium. Keratinocytes survived as contamination in PMA medium whereas PDBu medium had minimal keratinocytes. MTT assay showed that PDBu has higher proliferative induction capacity than PMA. In even lower concentration of PDBu in medium, melanocytes survived till 72 hours without significant cell loss in compared to PMA medium. CONCLUSION: PDBu can be a valuable replacement for PMA in human melanocyte culture. Higher proliferation induction, unfavourable to keratinocyte survival and less hydrophobicity make PDBu a promising alternative for quicker establishment of pure human melanocyte cultures especially in cosmetic in vitro experimental dermatology.

Effects of Lead in Various Preparatory Stages of Nagabhasma on Function and Histopathology of Cornu Ammonis of Hippocampus.

INTRODUCTION: Heavy metals are frequently used in the preparations of traditional/folk medicines. One such preparation in Ayurveda is Nagabhasma, in which lead is the main ingredient. Lead is non-essential element to the human body and is known toxic substance to many organ systems. However, it is claimed that, the highly toxic metallic lead will be converted into health beneficial organo-metallic compound when raw lead is subjected to various traditional methods of purification during preparation as mentioned in the ancient medicinal system. AIM: The present study is designed to evaluate the effect of such detoxification of lead in various stages of authentically prepared Nagabhasma on the learning and memory. MATERIALS AND METHODS: Using half of the human equivalent doses of traditionally prepared Nagabhasma, at intermittent stages of its preparation were fed orally to healthy Wistar rats for 30 days. After treatment, the immediate effect and residual effect after 2 months was evaluated by subjecting them to passive avoidance test. Then rats were sacrificed and hippocampus was collected for histopathological evaluation. RESULTS: Pure lead treated animals showed deficit in learning and memory which is indicated by spending more time in the dark compartment in passive avoidance test. However, animals treated with stage 1 to 4 Nagabhasma showed gradual increase in the memory and learning. This observation is substantiated by the findings of the histopathology of the Cornu Ammonis (CA) region of hippocampus. CONCLUSION: The results of the present study indicate that, the metallic toxicity of the lead used in the preparation of bhasma was gradually decreased from stage 1 to stage 4 of preparation. Therefore, the traditional way of preparing the metallic bhasma is very critical in eliminating the possible health hazardous metallic lead toxicity.