Efficacy of modified cell block cytology compared to fine needle aspiration cytology for diagnostic oral cytopathology.

Cell blocks and fine needle aspirations can be used for cytopathological diagnosis. Conventional fine needle aspiration smears provide limited material for diagnosis. The cell block technique provides more tissue, which improves diagnostic accuracy. We compared a modified cell block cytology to fine needle aspiration for providing optimal preservation of histochemical and immunocytochemical properties. We used 30 fine needle aspirates from oral lesions in two groups: group 1, fine needle aspiration cytology; group 2, cell block cytology. Smears of fine needle aspirates were stained with Papanicolaou. For the modified cell block technique, aspirated material was centrifuged to create a cell pellet, which then was fixed with Nathan alcohol formalin substitute. After routine histopathological processing, cell pellets were embedded in paraffin, then sectioned and stained with hematoxylin and eosin. Sections were compared to Papanicolaou stained smears of fine needle samples. Cellular morphology and staining quality of modified cell block samples were superior to fine needle aspiration cytology; both methods exhibited distinct nuclear morphology. Modified cell blocks provide excellent cytopathologic features compared to fine needle aspiration cytology.

Polyphenolic enriched extract of Cassia glauca Lamk, improves streptozotocin-induced type-1 diabetes linked with partial insulin resistance in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditionally Cassia glauca (CG) has been used to treat diabetes. AIM OF THE STUDY: The study was undertaken to evaluate anti-diabetic and antioxidant activity of polyphenolic enriched extract of CG in standardized streptozotocin (STZ)-induced diabetic rats. MATERIALS AND METHODS: The effect of ethanol (CGE) and water (CGW) extracts of CG (200 and 400mg/kg) treatment were evaluated in STZ (50mg/kg, iv) induced diabetic rats. On 10th day, oral glucose tolerance test and degree of insulin resistance was calculated. On 13th day, insulin tolerance test was performed to know the peripheral utilization of glucose. On 15th day, blood glucose, lipid profiles and endogenous antioxidant levels were estimated. In addition, the effects on oral glucose/sucrose tolerance test in normal rats. Further, HPLC fingerprinting profile of CGE and simultaneous quantification of biomarkers were carried out. RESULTS: Supplementation with CGE and CGW significantly reduced STZ-induced deleterious effects and improved glucose tolerance, and insulin tolerance. In addition, supplementation also decreased oxidative stress by improving endogenous antioxidant levels. Furthermore, administration significantly improves sucrose tolerance suggesting that extract possess inhibition of α-glucosidase enzyme. Further, HPLC studies revealed that CGE contains three bioactive polyphenolic compounds viz., rutin (0.10±0.01mg/g), luteolin-7-glucoside (0.06±0.01mg/g) and isorhoifolin (0.7±0.05mg/g). CONCLUSION: Observed beneficial outcome of CG might be attributed to the presence of polyphenolic compounds and mediated by interacting with multiple targets of diabetes and oxidative stress. Taken together, this study provided the scientific evidence for the traditional use of CG.

Adaptogenic and in vitro antioxidant activity of flavanoids and other fractions of Argyreia speciosa (Burm.f) Boj. in acute and chronic stress paradigms in rodents.

Argyreia speciosa (sweet) (Burm.f.) Boj. is an Ayurvedic rasayana plant used as an adaptogen. The present study reports the investigations done on the adaptogenic property of ethanol (EtAS; 100 and 200 mg/kg; po), ethyl acetate (EAAS; 100 and 200 mg/kg; po) fraction and flavanoids such as quercetin and kaempferol (25 mg/kg; po) of the root. Immobilization induced acute stress (AS; 3 days) and chronic stress (CS; 7 days) and swimming induced stress models were used to screen the anti-stress effect of the plant fractions and isolated flavanoids. The tested doses of EtAS and isolated flavanoids were able to produce significant effects in normalizing altered serum biochemical parameters and the severity of ulcer in both AS and CS models. Higher dose of EtAS, quercetin and kaempferol (25 mg/kg; po) were found to be significant in restoring the hypertrophy of adrenal gland and atrophy of spleen and thymus gland only in CS model. Greater swimming time was noted in the mice pretreated with tested doses of flavanoids and EtAS. In addition, levels of adrenal ascorbic acid and cortisol were restored compared to stress control group. EtAS exhibited significant scavenging effect of DPPH, hydroxyl radical and LPO. Thus, EtAS, quercetin and kaempferol are capable of increasing the capacity to tolerate non-specific stress in experimental animals, as evident from restoration of large number of parameters in the stress models studied. Bioactivity of EtAS may be due to the synergetic action of isolated flavanoids. Improvement in stress markers may be due its prolong effect of resistance to stress and partly due to free radical scavenging activity.

Preparation and characterization of novel semi-interpenetrating polymer network hydrogel microspheres of chitosan and hydroxypropyl cellulose for controlled release of chlorothiazide.

Novel semi-interpenetrating polymer network (IPN) hydrogel microspheres of chitosan (CS) and hydroxypropyl cellulose (HPC) were prepared by emulsion-cross-linking method using glutaraldehyde (GA) as a cross-linker. Chlorothiazide (CT), a diuretic and anti-hypertensive drug with limited water solubility, was successfully encapsulated into IPN microspheres. Various formulations were prepared by varying the ratio of CS and HPC, percentage drug loading and amount of GA. Microspheres were characterized by Fourier transform infrared (FTIR) spectroscopy to investigate the formation of IPN structure and to confirm the absence of chemical interactions between drug, polymer and cross-linking agent. Scanning electron microscopy (SEM) was performed to study the surface morphology of the microspheres. SEM showed that microspheres have smooth surfaces. Particle size, as measured by laser light scattering technique, gave an average size ranging from 199-359 mum. Differential scanning calorimetry (DSC) was performed to know the formation of IPN structure. X-ray diffraction (X-RD) studies were performed to understand the crystalline nature of the drug after encapsulation into IPN microspheres. Encapsulation of drug up to 76% was achieved as measured by UV spectroscopy. Both equilibrium and dynamic swelling experiments were performed in 0.1 N HCl. Diffusion coefficients (D) for water transport through the microspheres were estimated using an empirical equation. In vitro release studies indicated the dependence of release rate on the extent of cross-linking, drug loading and the amount of HPC used to produce the microspheres; slow release was extended up to 12 h. The release data were also fitted to an empirical equation to compute the diffusional exponent (n), which indicated that the release followed the non-Fickian trend.

Co-crystals: a novel approach to modify physicochemical properties of active pharmaceutical ingredients.

Crystal form can be crucial to the performance of a dosage form. This is especially true for compounds that have intrinsic barriers to drug delivery, such as low aqueous solubility, slow dissolution in gastrointestinal media, low permeability and first-pass metabolism. The nature of the physical form and formulation tends to exhibit the greatest effect on bioavailability parameters of water insoluble compounds that need to be given orally in high doses. An alternative approach available for the enhancement of drug solubility, dissolution and bioavailability is through the application of crystal engineering of co-crystals. The physicochemical properties of the active pharmaceutical ingredients and the bulk material properties can be modified, whilst maintaining the intrinsic activity of the drug molecule. This article covers the advantages of co-crystals over salts, solvates (hydrates), solid dispersions and polymorphs, mechanism of formation of co-crystals, methods of preparation of co-crystals and application of co-crystals to modify physicochemical characteristics of active pharmaceutical ingredients along with the case studies. The intellectual property implications of creating co-crystals are also highly relevant.

Sequential interpenetrating polymer network hydrogel microspheres of poly(methacrylic acid) and poly(vinyl alcohol) for oral controlled drug delivery to intestine.

Sequential interpenetrating networks of poly(methacrylic acid) and poly(vinyl alcohol) have been prepared and cross-linked with glutaraldehyde to obtain pH sensitive microspheres by a water-in-oil emulsification method. Microspheres have been used to deliver the chosen model anti-inflammatory drug viz., ibuprofen to the intestine. Ibuprofen was encapsulated up to 70% within polymeric matrices. The interpenetrating polymer network formed was analysed by Fourier transform infrared spectroscopy. Differential scanning calorimetry and X-ray diffraction analyses were done on drug-loaded microspheres to confirm the polymorphism of ibuprofen. Results of this study indicated the molecular level dispersion of ibuprofen in the developed microspheres. Scanning electron microscopy confirmed the spherical nature and smooth surfaces of the microspheres produced. Mean particle size of the microspheres as measured by laser light scattering ranged between 51-176 microm. Swelling was performed in the simulated gastric as well as the intestinal conditions. Microspheres showed a pulsatile swelling behaviour when pH of the swelling media was altered. The swelling data have been fitted to an empirical equation to understand water transport trends as well as to calculate the diffusion coefficients. Values of diffusion coefficients in acidic media were lower than those found in the basic media. Values of diffusion coefficients decrease with increasing cross-linking of the matrix. In vitro release studies have been performed in 1.2 and 7.4 pH media to simulate the gastric and intestinal conditions. The in vitro release results indicated a dependence on the pH of the release media, extent of cross-linking and the amount of drug loading. The release data were fitted to an empirical relation to estimate the transport parameters and thereby to understand the transport mechanism.

Benign fibrous histiocytoma of bone: a case report.

Benign fibrous histiocytoma (BFH) occurs commonly in soft tissues with very few cases of BFH of bone on record. We report a case of BFH of tibia in fifty year old male who presented with supracondylar fracture femur due to trivial fall and a brief review of literature.

Guinea worm infection of urinary bladder manifesting as obstructive uropathy in rural Maharashtra.

Guinea worm or Dracunculus medinensis is a well-documented helminthic infestation in many areas of Asia. In this report, we describe a rare case of guinea worm infestation in a 25-year-old woman who had developed symptoms of obstructive uropathy, in whom fragments of guinea worm were removed after urethral catheterization. To the best of our knowledge, adult guinea worm occurring in the urinary bladder has not been previously described.

A new method for radiochemical separation of arsenic from irradiated germanium oxide.

Radioarsenic labelled radiopharmaceuticals could be a valuable asset to Positron Emission Tomography (PET). In particular, the long half-lives of (72)As (T(1/2)=26 h) and (74)As (T(1/2)=17.8 d) allow to investigate slow physiological or metabolical processes, like the enrichment and distribution of antibodies in tumor tissue. This work describes the direct production of no-carrier-added (nca) arsenic isotopes *As, with *=71, 72, 73, 74 or 77, the reaction to [*As]AsI(3) and its radiochemical separation from the irradiated solid germanium oxide via polystyrene-based solid-phase extraction. The germanium oxide target, irradiated at a cyclotron or a nuclear reactor, is dissolved in concentrated HF and Ge is separated almost quantitatively (99.97%) as [GeF(6)](2-). [*As]AsI(3) is formed by addition of potassium iodide. The radiochemical separation yield for arsenic is >90%. [*As]AsI(3) is a versatile radioarsenic labelling synthon.

Prevalence of seroreactivity among blood donors in rural population.

The seroreactivity pattern amongst blood donors in rural population was studied at S.R.T. Rural Medical College and Hospital, Ambajogai (M. S.). The study period was from January 1996 to December 2001. A total number of 12,240 blood donors were screened. The voluntary donation was 36.98% and replacement donors were 63.02%. No professional donor is bled in our blood bank. The HIV seroreactivity among voluntary donors was 1.56% and 2.11% in replacement donors. The HBsAg seroreactivity was 2.78% in voluntary donors and 4.84% in replacement donors. VDRL seroreactivity is 1.12% in replacement donors. No malarial parasite and HCV seroreactive donor was found in our study period. We have found the magnitude of hepatitis to be far more than that of HIV. Hence testing for HCV routinely is mandatory, besides HBsAg.

Transient adhesion of platelets in pump-oxygenator systems: influence of SMA and nitric oxide treatments.

We employed gamma scintigraphy to quantify the transient accumulations of platelets in pump-oxygenator systems employed in cardiopulmonary bypass (CPB). A flat sheet microporous polypropylene membrane oxygenator (Cobe Duo) was employed, with and without siloxane/caprolactone oligomer coating (SMA) (n = 8 each). The effect of nitric oxide gas infusion on platelet deposition was also evaluated for the uncoated Cobe Duo system (n = 10 each). Scintigraphic images of radiolabelled cells were obtained and converted to numbers of all platelets, labeled and unlabeled, adhering to the pump and oxygenator surfaces. These numbers were compared, by study group, for a 90-min period of normothermic CPB in the adult pig, employing standard prime and anticoagulation regimens. Platelets adhered in large numbers to control oxygenators, reaching maxima (> 20% of the circulating platelet mass) 30 min following institution of CPB, and decreasing for the duration of CPB. SMA treatment significantly decreased platelet adhesion following a 5-10-min transient accumulation period. Nitric oxide infusion significantly reduced platelet adhesion throughout the CPB period. Platelet accumulations on the high fluid shear centrifugal pump surfaces increased monotonically to maxima at about the same time as for the oxygenators, but did not decrease thereafter. Higher platelet surface densities were observed on the centrifugal pump surfaces than on the oxygenator surfaces. CPB with the untreated circuit tended to reduce circulating platelet counts vs theoretical values based on hemodilution alone. In contrast, SMA significantly increased the circulating platelet count versus the untreated control group. These results indicate that platelet adherence to the foreign surfaces of CPB equipment are influenced in characteristic ways by time and fluid shear. SMA treatment and nitric oxide infusion both reduce platelet adhesion to oxygenator surfaces. SMA treatment spares these cells for the circulation.

Development of novel 19F NMR pH indicators: synthesis and evaluation of a series of fluorinated vitamin B6 analogues.

We have synthesized a series of novel fluorinated vitamin B6 analogues (6-fluoropyridoxol derivatives) as potential 19F NMR pH indicators for use in vivo. Modifications included addition of aldehyde, carboxyl or aminomethyl groups at the 4- or 5-ring position, and examination of a trifluoromethyl moiety as an internal chemical shift standard. The variation in chemical shift with respect to acid-base titration showed pKa values in the range 7.05-9.5 with a chemical shift sensitivity in the range 7.4-12 ppm. Several of the molecules readily cross cell membranes providing estimates of both intra- and extra-cellular pH in whole blood. 6-Fluoropyridoxamine (6-FPAM) exhibits a pKa = 7.05, which is closer to normal physiological pH than the parent molecule 6-fluoropyridoxol (6-FPOL) (pKa = 8.2), and should thus, be useful for precise and accurate measurements of pH in vivo. Enhanced spectral resolution for 6-FPAM over 6-FPOL is demonstrated in whole blood and the perfused rat heart.

SPECT electronic collimation resolution enhancement using chi-square minimization.

An electronic collimation technique is developed which utilizes the chi-square goodness-of-fit measure to filter scattered gammas incident upon a medical imaging detector. In this data mining technique, Compton kinematic expressions are used as the chi-square fitting templates for measured energy-deposition data involving multiple-interaction scatter sequences. Fit optimization is conducted using the Davidon variable metric minimization algorithm to simultaneously determine the best-fit gamma scatter angles and their associated uncertainties, with the uncertainty associated with the first scatter angle corresponding to the angular resolution precision for the source. The methodology requires no knowledge of materials and geometry. This pattern recognition application enhances the ability to select those gammas that will provide the best resolution for input to reconstruction software. Illustrative computational results are presented for a conceptual truncated-ellipsoid polystyrene position-sensitive fibre head-detector Monte Carlo model using a triple Compton scatter gamma sequence assessment for a 99mTc point source. A filtration rate of 94.3% is obtained, resulting in an estimated sensitivity approximately three orders of magnitude greater than a high-resolution mechanically collimated device. The technique improves the nominal single-scatter angular resolution by up to approximately 24 per cent as compared with the conventional analytic electronic collimation measure.

Analytic treatment of resolution precision in electronically collimated SPECT imaging involving multiple-interaction gamma rays.

The widely applied single-interaction analytic expression characterizing the energy resolution component of the angular resolution precision for an electronically collimated point source is extended to include multiple-interaction Compton scatter sequences as well as sequences terminated by photoelectric absorption. The analytic formulation is developed using the statistical variance of the mean for components comprising composite, multivariate resolution precision estimators. It is demonstrated that enhanced resolution precision in the incident interaction scatter angle is attained when use is made of information from multiple interactions. An improvement in the resolution precision of up to approximately 40% is observed for triple Compton scatter. Comparison of the analytic estimates with Monte Carlo/chi-square results shows good agreement.

Simultaneous intracellular and extracellular pH measurement in the heart by 19F NMR of 6-fluoropyridoxol.

6-Fluoropyridoxol (6-FPOL) was evaluated as a simultaneous indicator of intracellular and extracellular pH and, hence, pH gradient in perfused rat hearts. After infusion, 19F NMR spectra rapidly showed two well-resolved peaks assigned to the intracellular and extracellular compartments, and pH was calculated on the basis of chemical shift with respect to a sodium trifluoroacetate standard. To demonstrate use of this molecule, dynamic changes in myocardial pH were assessed with a time resolution of 2 min during respiratory and metabolic alkalosis or acidosis and ischemia. For a typical heart, intracellular pH (pHi) = 7.14+/-0.01 and extracellular pH (pHe) = 7.52+/-0.02. In response to metabolic alkalosis, pHi remained relatively constant and the pH gradient increased. In contrast, respiratory challenge caused a significant increase in pHi. Independent measurements using pH electrodes and 31P NMR confirmed validity of the 19F NMR results.

Influence of cardiopulmonary bypass on platelet and neutrophil accumulations in internal organs.

The authors employed gamma scintigraphy to quantify the post bypass accumulations of platelets and neutrophils in the lung, liver, and heart of adult pigs subjected to a standard 90 min regimen of normothermic cardiopulmonary bypass (CPB). Coated and uncoated microporous polypropylene oxygenator circuits were studied for Cobe Duo (Arvada, CO) oxygenators (amphophilic silicone-caprolactone oligomer [SMA] coating, n = 8 each) and Medtronic Maxima (Irvine, CA) oxygenators (Carmeda heparin coating, n = 5 each). Images of cells in the organs (deposited + blood pool) were corrected for tissue absorption and other factors and compared for a 2 hr period post CPB, using repeat measures ANOVA and rank tests. Platelet accumulations in internal organs correlated positively with whole blood platelet counts and negatively with platelet deposits in oxygenators during CPB. In general, uncoated CPB circuits significantly reduced platelet and neutrophil accumulations in lung, liver, and heart versus preCPB controls for the post CPB interval, for both systems. The SMA treatment significantly increased platelet accumulations versus uncoated controls in lung, liver, and heart for the 2 hr period, including the majority of the post CPB sampling intervals; platelet densities did not reach preCPB levels. Neutrophil accumulations were unaffected by the SMA coating. Carmeda heparin treatment significantly increased platelet accumulations in the liver, but not lung or heart. Despite preservation of circulating neutrophils observed with the Carmeda heparin treatment, neutrophil accumulations in internal organs were not elevated post CPB.

6-Fluoropyridoxal polymer conjugates: novel 19F pH indicators for magnetic resonance spectroscopy.

Fluorinated macromolecular probes (6-fluoropyridoxal-polymer conjugates) have been synthesized and characterized as potential pH indicators for magnetic resonance spectroscopy and imaging applications. The 19F pH sensor 2-fluoro-5-hydroxy-3-(hydroxymethyl)-6-methyl-4-pyridinecarboxal-+ ++dehyde (6-fluoropyridoxal; 2) has been conjugated to carrier molecules (polyamino dextran, polylysine, and albumin) by reductive alkylation for enhanced vascular retention and tissue targeting. The pH indicator polymer conjugates were purified by exhaustive dialysis and isolated in good yields (66-84%). The 6-fluoropyridoxal-polymer conjugates exhibit excellent 19F pH sensitivity and pKa suitable for in vivo studies. The potential application of these polymeric indicators has been demonstrated in whole blood. These novel macromolecular pH probes offer a new approach for studying tissue physiology.

Platelet and neutrophil distributions in pump oxygenator circuits. III. Influence of nitric oxide gas infusion.

The authors used quantitative gamma scintigraphy to evaluate the influence of nitric oxide gas on platelet and neutrophil deposition in Cobe Duo microporous oxygenators during cardiopulmonary bypass (CPB). The effects of nitric oxide gas on circulating platelet and neutrophil counts and platelet function also were assessed. Animals were prepared by standard methods. Cells were harvested, labeled (111 In platelet and 99mTc neutrophil), infused, and recirculated. Nitric oxide gas, a guanylate cyclase pathway promoter, was infused int he Duo gas port at 500 ppm (t = 0-60 min), increased to 1,000 (t = 60-80 min), and stopped (final, 10 min). Images were taken at 10-15 min intervals during CPB. Standard isotope image corrections were made. No differences between nitric oxide gas and control experiments were observed for flow, pressure, hematocrit, or replacement volume. Nitric oxide gas infusion significantly (p < 0.05) reduced both platelet adherence to the oxygenator and in vitro platelet aggregation. Neutrophil adhesion tended to be lower, and circulating platelet and neutrophil counts tended to be higher with nitric oxide gas infusion. Results of in vitro aggregometry studies using rabbit platelets indicate that the class V phosphodiesterase inhibitor zaprinast can strongly enhance the inhibitory effects of nitric oxide. The authors conclude nitric oxide gas is a promising platelet sparing agent in the setting of CPB.

Profile of malignancies at Medical College, Ambajogai--(15 years retrospective study).

Present study comprises of a preliminary analysis of malignant lesions, diagnosed and confirmed by histopathological examination, carried out in Department of Pathology, S. R. T. R. Medical College, Ambajogai, from August 1976 to July 1991. 30,168 specimens were received for histopathological examination out of which 2,663 cases were diagnosed as malignant lesions (8.82%). 163 cases of leukemias were diagnosed by peripheral smear and bone marrow examination. Our of 2826 patients with malignant lesions 1,566 (55.41%) were males and 1,260 (44.59%) were females with M:F ratio as 1.24:1. Analysis of all malignant lesions showed more predilection towards genital organs (26.22%), gastrointestinal tract (13.72%) and breast (10.47%). Penis (11.36%), gastrointestinal tract (16.73%), oral cavity and tongue with pharynx (20.81%) and skin (11.11%) were leading sites in males where as cervix (30.63%) breast (22.46%), gastrointestinal tract (10.00%), skin (6.82%) and lymphoma, leukaemias (7.30%) were common sites for malignancies in females. The higher incidence of genital and skin tumours is discussed in the paper. Available data is compared with data from Tata Memorial Cancer Hospital Bombay and Cancer Hospital at Barshi in Solapur District of Maharashtra.