Tinospora cordifolia, a novel source of extracellular disaccharidases, useful for human disaccharidase deficiency therapy.

Disaccharide intolerance is the inability to digest certain carbohydrates due to a lack of one or more intestinal disaccharidases (e.g., lactase, maltase, isomaltase and sucrase). Symptoms include diarrhea, abdominal distention and flatulence. Management of the disorder by external enzymes supplementation has not yet been attempted. We report that the medicinal plant Tinospora cordifolia contains substantial amounts of all disaccharidases required for intestinal digestion of carbohydrates. The plant is also a rich source of saccharifying amylase. We recovered (units/100 g fresh stem) amylase: 49,000+500, maltase: 400+50, isomaltase: 130+50, sucrase: 4500+500, acid lactase: 350+30, cellobiase: 35+10 and trehalase: 40+10 by buffer extraction of the blended stem. Crude enzymes in the forms of stem powder, lyophilized aqueous extract and ethanol precipitated protein were found to be stable. Disaccharidases were optimally active at 50 (0) C in the pH range of 4-5. Lactase was an acid lactase similar to the type linked with human lactose intolerance. Enzymes were catalytically stable in the pH range of 2-7 and temperature range of up to 40 (0) C. T. cordifolia enzyme was non-toxic up to a dose of 200 mg protein/kg body weight.

Characterization of nimbidiol as a potent intestinal disaccharidase and glucoamylase inhibitor present in Azadirachta indica (neem) useful for the treatment of diabetes.

Azadirachta indica, used in antidiabetic herbal drugs, was reported to contain α-glucosidase inhibitor. Bioassay guided purification characterized the inhibitor as nimbidiol (a diterpenoid), present in root and stem-bark of the tree. Nimbidiol inhibited intestinal (mammalian) maltase-glucoamylase, sucrase-isomaltase, lactase, trehalase and fungal α-glucosidases. Nimbidiol showed a mixed competitive inhibition on intestinal carbohydrases. IC50, Ki and Ki' (µM) were 1.35 ± 0.12, 0.08 ± 0.01, 0.25 ± 0.11, respectively, for maltase-glucoamylase (maltotetraose as substrate). Nimbidiol was more potent inhibitor of isomaltase (IC50 0.85 ± 0.035 µM), lactase (IC50 20 ± 1.33 µM) and trehalase (IC50 30 ± 1.75 µM) than acarbose, voglibose, salacinol, kotalanol and mangiferin. Ki and Ki' values (µM) for intestinal sucrase were 0.7 ± 0.12 and 1.44 ± 0.65, respectively. Development of nimbidiol as an antidiabetic drug appears to be promising because of broad inhibition spectrum of intestinal glucosidases and easy synthesis of the molecule.

Anatomical variations of uncinate process observed in chronic sinusitis.

Chronic Sinusitis, an extremely persistent illness, is surgically best treated by Functional Endoscopic Sinus Surgery. The ostiomeatal complex is the main area targeted and within it uncinate process is the first anatomical structure encountered. The significance of anatomical variations concerning age and sex of uncinate process in chronic sinusitis were evaluated. A prospective study on 50 patients of chronic sinusitis (100 uncinate processes) was done. The results were tabulated and analyzed using Statistical Package for Social Science (SPSS) 16.0. Type I superior attachment of uncinate process (67 %) was the most common variety in all ages and both sexes and a statistically significant relationship between Type I superior attachment of uncinate process and sex was found (p < 0.05). The typical uncinate process was most common (70 %) followed by medial deviation of the uncinate (24 %). This difference in occurrence was significant with respect to both age and sex (p < 0.05). Anatomical variations of uncinate process are not responsible for causing chronic sinusitis. Mere presence of these variations of uncinate is not an indication for FESS.

Pattern of Nonmelanoma Skin Cancers in Sikkim, India: A 3-year Clinicopathological Review.

Basal cell carcinoma (BCC) and Squamous cell carcinoma (SCC), in combination, are referred to as Nonmelanoma skin cancers (NMSCs). NMSC has not been extensively studied in the population of the North eastern India as it has been done in the rest of India. This study was performed to understand the pattern of NMSCs in head and neck region in Sikkimese population. The present study is a retrospective analysis of NMSC of head and neck region that presented to the Department of ENT, Sikkim Manipal Institute of Medical sciences (SMIMS), a tertiary care center in Gangtok, Sikkim from July 2009 to July 2012. They were reviewed according to age, gender, site of involvement and histological types. Of the thirty-seven cases of NMSC analyzed during the 3 year study period, 24 cases were SCC and 13 cases were BCC. The most common age group for SCC was 40-50 years and BCC was 61-70 years. The most common site for both SCC and BCC was the upper lip. For SCC, the most common histologically type was well-differentiated and for BCC, it was the solid type. NMSCs comprise a small but significant proportion of all skin cancer patients in the world. SCC is the commonest variety unlike in western countries. The importance of understanding the risk factors as well as studying the frequency of histopathological patterns can help in the overall prognostic outlook for patients and also in planning an effective management.

Total Reconstruction of Lower Eyelid in a Post-traumatic Patient Using Modified Fricke's Cheek Flap.

Eyelids are very complex structure, reconstruction of which is a challenge to surgeons. Reconstruction of eyelids may be required in a variety of conditions like congenital anomalies, trauma, or postsurgical excision in malignant lesions involving the eyelids. There are numerous ways to reconstruct the eyelids; the best procedure depends on both the skill of the surgeon and the condition of the patient. Fricke' lateral temporal based flap was first described in 1829 for reconstruction of the eyelids and lateral canthal region. This flap had inherent problems regarding cosmetic appearance of the eyebrows. The modified Fricke's flap based on the cheek has the advantage of avoiding such complications. It is very easy and rapid outpatient department (OPD) based procedure with acceptable cosmetic and functional result. It can be done by all ear, nose, and throat (ENT) and head and neck surgeons without any reconstructive training. In this article we are presenting a case of total reconstruction of lower eyelid using the modified Fricke's cheek flap.

Persistent deciduous molar as an etiology for a maxillary sinus mucocele.

Mucoceles are accumulations of trapped mucus, forming cystic expansile lesions. Maxillary sinus mucoceles are rare amongst paranasal sinus mucoceles, usually being a late sequel of Caldwell-Luc surgery. We present a case of a maxillary sinus mucocele due to a persistent carious deciduous molar in a middle aged patient, reported to highlight its unusual etiology.

Fine needle aspiration cytology as the primary diagnostic tool in thyroid enlargement.

BACKGROUND: In the preoperative decision-making of the thyroid swellings, fine needle aspiration cytology (FNAC) is becoming an ever more vital tool. OBJECTIVES: To compare the advantage of preoperative FNAC of thyroid swellings with postoperative histopathology to reach a consensus protocol as a simple procedure for diagnosis and optimal management of thyroid swellings. MATERIALS AND METHODS: A prospective study of preoperative FNAC was carried out on 178 incidental thyroid swellings attending a tertiary care centre in Kishanganj, Bihar. Evidence-based surgical interventions were done, irrespective of FNAC findings and diagnosis was confirmed by histopathological examination (HPE) postoperatively in all the cases. RESULTS: In the FNAC, preponderance of the cases (75.84%) was colloid goitre followed by granulomatous thyroiditis; follicular carcinoma was noted in 7.30 percent and anaplastic carcinoma in 3.37 percent of cases. Histopathological examination showed colloid goitre predominantly (76.97%), followed by follicular carcinoma (8.99%). The overall prevalence of malignancy was 11.24 percent diagnosed by HPE and 9.55 percent by FNAC. In our FNAC series sensitivity of was 90 percent while specificity was 100 percent; accuracy was 98.88 percent. Predictive value of a positive test and negative tests was 100 percent and 98.75 percent respectively. CONCLUSION: The study highlights that FNAC should be treated as a first-line diagnostic test for thyroid swellings to guide the management though this is not a substitute for HPE as a need to improve primary healthcare in India.

Purification and characterization of a thiol amylase over produced by a non-cereal non-leguminous plant, Tinospora cordifolia.

A 43kDa α-amylase was purified from Tinospora cordifolia by glycogen precipitation, ammonium sulfate precipitation, gel filtration chromatography, and HPGPLC. The enzyme was optimally active in pH 6.0 at 60°C and had specific activity of 546.2U/mg of protein. Activity was stable in the pH range of 4-7 and at temperatures up to 60°C. PCMB, iodoacetic acid, iodoacetamide, DTNB, and heavy metal ions Hg(2+)>Ag(+)>Cd(2+) inhibited enzyme activity while Ca(2+) improved both activity and thermostability. The enzyme was a thiol amylase (3 SH group/mole) and DTNB inhibition of activity was released by cysteine. N-terminal sequence of the enzyme had poor similarity (12-24%) with those of plant and microbial amylases. The enzyme was equally active on soluble starch and amylopectin and released maltose as the major end product.

An attempt to define the type of biopsy in a sinonasal lesion showing bony erosion.

OBJECTIVES: To present a case of sino-nasal destructive mass initially diagnosed as an inflammatory lesion following punch biopsy from the lesion however the post surgical histopathology was diagnostic of Grade 2 angiocentric immunoproliferative lesion (AIL). The reasons for the initial misdiagnosis are analyzed. MATERIALS AND METHODS: A 76-year-old male patient presenting with progressive bilateral nasal obstruction for 1 year. Repeated punch biopsies from the mass were suggestive of an inflammatory lesion. RESULT: The patient underwent surgical exenteration of the mass and the final histopathology report suggested AIL Grade 2. The patient was thereafter treated with chemotherapy and radiotherapy. CONCLUSION: Initial superficial punch biopsies lead to incorrect diagnosis leading to an unnecessary surgical exenteration. The explanations for the initial misdiagnosis are given below and appropriate diagnostic protocols, mode and depth of biopsy are suggested based on the case study.

Purification and characterization of a low molecular weight endo-xylanase from mushroom Termitomyces clypeatus.

A low molecular weight endo-xylanase (EC 3.2.1.8) was purified from an edible mushroom Termitomyces clypeatus grown in submerged medium with oat spelt xylan. Xylanase was purified to apparent homogeneity by ammonium sulfate fractionation and gel filtration chromatography. Its molecular weight was determined by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 12 kDa. The enzyme was found to be most active at 50 degrees C and pH 5.0, being most stable at pH 6.5. The K(m) for oat spelt xylan was determined to be 10.4 mg/ml. The specificities of the enzyme was observed to be highly specific towards oat spelt xylan and was inhibited by mercuric chloride (HgCl(2)), N-bromosuccinimide, and trans-1,2-diaminocyclohexane-N',N',N',N'-tetraacetic acid strongly. The inhibitory action of N-bromosuccinimide on enzyme confirmed the presence of one tryptophan residue in its substrate-binding site. Amino acid analysis for xylanase showed the presence of high amount of hydrophobic serine, glycine, threonine, and alanine residues. The N-terminal sequencing study for the previously purified and characterized 56 kDa xylanolytic amyloglucosidase reveal the presence of 33.30% identity with glucoamylase chain A from Aspergillus awamori. The N-terminal sequence analysis of the present 12 kDa enzyme showed highest similarity (72.22% identity) towards xylanase from Neurospora crassa.

Clinicopathological correlates of pediatric head and neck cancer.

BACKGROUND: The spectrum of head and neck tumors in children continues to be the cause of diverse, diagnostically challenging issues. AIMS: To demonstrate and compare the unique clinicopathological features in our study population and their correlations with the final histopathological diagnosis. METHODS: Fifty-three children with head and neck cancer were examined thoroughly at the Otorhinolaryngology department in a tertiary care teaching hospital followed by histopathological studies. RESULTS: Lymphomas were the most common malignant lesions seen followed by rhabdomyosarcomas, nasopharyngeal carcinomas, and others like thyroid carcinomas and eosinophilic granulomas. In the neck, the commonest cause of primary malignant disease was lymphoma; however, the most frequent lesion was reactive lymphadenitis. In the sinonasal region, the commonest malignancy was rhabdomyosarcoma, which often had extension to the orbit and the face. Recurrent epistaxis was found universally in the malignant cases of this region. In the facial region, disfiguring swelling with proptosis was mainly caused by rhabdomyosarcoma. The only case of tonsillar malignancy was due to non-Hodgkin lymphoma. The duration of disease was less than 1 year. CONCLUSION: The most common manifestation of the malignant lesions in the pediatric age group was with a history of an enlarging, painless neck swelling. Still, an insignificant lump in the neck or recurrent bleeding from nose may be the manifestation of an underlying cancer.

Hypoglycemic activity of the antioxidant saponarin, characterized as alpha-glucosidase inhibitor present in Tinospora cordifolia.

Tinospora cordifolia, used in anti-diabetic herbal drug preparations, was reported [12] to contain an alpha-glucosidase inhibitor, characterized as saponarin (apigenin-6-C-glucosyl-7-O-glucoside). The leaf extract had appreciable antioxidant and hydroxyl radical scavenging activities and contained the flavonoid in the range of 32.1 +/- 1.5-45.5 +/- 3.5 mg/g of dry solid. Saponarin showed mixed competitive inhibition on activities of alpha-glucosidase and sucrase of different origins. IC(50), Ki and ki' values determined were 48 muM, 8 muM and 19.5 microM respectively for intestinal maltase and 35 microM, 6 microM and 13 microM respectively for intestinal sucrase. When given orally to maltose-fed rat, saponarin showed hypoglycemic activity in the range of 20-80 mg/kg compared to 100-200 mg/kg for acarbose as reported.

Fourth branchial pouch sinus - an unusually late initial presentation.

OBJECTIVE: Here we present an unusual case of a male patient with 4th branchial pouch sinus with an initial presentation of pseudothyroiditis at an unusually late age of 20 years. METHOD: Case report and a review of the world literature concerning 4th branchial pouch sinus is presented. RESULT: The patient was diagnosed by radiology and endoscopy and treated surgically. Histopathology revealed chronic inflammatory changes, which pointed towards recurrent sub clinical low-grade inflammations over the years keeping the condition occult till 20 years of age. CONCLUSION: According to available literature it almost always presents in the first decade of life when it might be misdiagnosed. However our report shows it may also exist with sub clinical inflammations or infections and present at a much later stage in life, when the possibility of such a sinus shouldn't be ruled out on the basis of lack of history of recurrent neck inflammations.

Spectrum of head and neck cancer in children.

AIM: To assess the incidence of different head and neck cancers in pediatric age group in a referral hospital. METHODS: In this prospective study, children below the age of 12 years underwent a thorough clinical, ENT examination and the diagnosis was conformed histologically in all the cases. RESULTS: Fifty-three were diagnosed to be suffering from different head and neck neoplasms among 21,216 children (0.25%). Male-to-female sex distribution was 1.78:1. The lymphomas were the most common (43.39%) followed by the rhabdomyosarcoma (20.75%) and the nasopharyngeal carcinoma (15.09%). Of the lymphomas, the non-Hodgkin's lymphoma was predominant (26.41%). Other lesions were thyroid carcinomas and mucoepidermoid carcinoma of parotid. CONCLUSIONS: Malignancy should always be considered in the differential diagnosis of masses in the head and neck region in children.

Occult foreign bodies in the aero digestive tracts in infancy.

Foreign body impaction in aero digestive tract in infancy is difficult to diagnose sometimes. In this review of five unusual cases of occult foreign body impaction in aero digestive tract has been presented and their management discussed.

Behavior disparities towards blood donation in Sikkim, India.

BACKGROUND: The aim of the current research was to determine disparities in blood donation motives among the general mass of Sikkim. AIMS: To identify the reasons for people donating and not donating blood voluntarily. SETTINGS AND DESIGN: Population based cross-sectional study in Gangtok, East Sikkim. PARTICIPANTS: 300 adults by two-stage cluster sampling technique. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Correlates of attitudes towards blood donation. DATA COLLECTION PROCEDURE: The data collection tool used for the study was a pre-tested structured interview schedule by which the principal investigator collected the data using interview technique. STATISTICAL ANALYSIS USED: Percentages and ODDS ratio were used in this study. RESULTS AND CONCLUSIONS: Out of 300 respondents, overwhelming majority (78.7%) of the respondents in the present study felt that people donate blood to save a friend or a relative. On the contrary, minority respondents (46%) were ready to donate blood voluntarily. Only 12.7% of the respondents had ever donated blood while 87.3% had never donated. Among ever donors, gender wise men donors were found to be more; 89% were married, half were from the 30 to 39 years age group. As the per-capita income or level of education increased, so did the percent of blood donors.

Benign calcified thyroid cyst with skin sinus formation.

OBJECTIVES: To present a rare case of calcified thyroid cyst with discharging neck sinus and its probable explanation. MATERIALS AND METHODS: A 75-year-old male patient presented to the outpatients department who had had thyroid swelling for the last 50 years and a discharging neck sinus from the swelling with ulceration and fungation of the adjoining area for the last year. RESULTS: An X-ray of the soft tissue of the neck showed tracheal shift to the right and irregular calcification of the left lobe of the thyroid. Fine needle aspiration cytology (FNAC) of the left lobe yielded little material with inflammatory cells as the swelling was too hard to be easily penetrated with a needle. FNAC of the right lobe revealed a picture similar to adenomatous goiter. A subtotal thyroidectomy was performed and the histopathologic report was suggestive of multinodular goiter with dystrophic calcification. CONCLUSION: Though the clinical features of the patient mimicked malignant change in a pre-existing adenomatoid goiter, histopathologically, it was found to be a calcified thyroid cyst. Probable explanation is given in the "Discussion" section.

Conservation of average hydrophobicity of apolar aminoacids in polypeptides constituting same glycosyl hydrolase sub-family enzymes.

Polypeptides constituting the same functional enzyme in cells of different origins have small sequence similarities among themselves. Amino acid analysis reveals that each glycosyl hydrolase sub-family polypeptides conserves an average hydrophobicity value for total constituent apolar amino acids. The value may be a measure of the driving force present in the polypeptide for designed primary collapse for three-dimensional active site formation.

Extracellular trehalose utilization by Saccharomyces cerevisiae.

Two haploid strains of Saccharomyces cerevisiae viz. MATalpha and MATa were grown in glucose and trehalose medium and growth patterns were compared. Both strains show similar growth, except for an extended lag phase in trehalose grown cells. In both trehalose grown strains increase in activities of both extracellular trehalase activities and simultaneous decrease in extracellular trehalose level was seen. This coincided with a sharp increase in extracellular glucose level and beginning of log phase of growth. Alcohol production was also observed. Secreted trehalase activity was detected, in addition to periplasmic activity. It appeared that extracellular trehalose was hydrolyzed into glucose by extracellular trehalase activity. This glucose was utilized by the cells for growth. The alcohol formation was due to the fermentation of glucose. Addition of extracellular trehalase caused reduction in the lag phase when grown in trehalose medium, supporting our hypothesis of extracellular utilization of trehalose.

Regulation of trehalose metabolism by Adox and AdoMet in Saccharomyces cerevisiae.

Effect of a potent methylation inhibitor oxidized adenosine (Adox), and a universal methyl group donor S-adenosyl-L-methionine (AdoMet) on trehalose metabolism was studied in two haploids of S. cerevisiae of mating types MATalpha, met3 (6460 -8D) and MATa, leu2, ura3, his4 (8534 -10A). Trehalose level decreased in presence of Adox in both strains. Both neutral trehalase (NT) and trehalose-6-phosphate (tre-6-p) synthase activities increased in presence of Adox in -8D strain. Decrease in trehalose level in -8D thus could not be explained in the light of increased tre-6-p synthase activity; however, it could be correlated with increased NT activity. In strain -10A, NT activity was reduced in presence of Adox while tre-6-p synthase activity increased. Enzyme activity profiles in -10A thus do not explain the reduced trehalose level on Adox treatment. Effect of AdoMet was not very prominent in either strain, though in -8D a small increase in trehalose level was seen on treatment. Intracellular AdoMet level of untreated cells of -10A was seen to be almost six times higher than that of -8D. Further, AdoMet treatment caused increase in its level compared to untreated cells, suggesting AdoMet uptake. No effect of either compound was seen on acid trehalase (AT) activity in any strain. The results suggest that there was a possible effect of demethylation on trehalose metabolism (particularly in the synthetic direction) in both strains, though effect of methylation was not very prominent, the reason for which is not very clear.