Augmentation of humoral and cellular immunity in response to Tetanus and Diphtheria toxoids by supercritical carbon dioxide extracts of Hippophae rhamnoides L. leaves.

Hippophae rhamnoides L. commonly known as Seabuckthorn (SBT), a wild shrub of family Elaegnacea, has extensively used for treating various ailments like skin diseases, jaundice, asthma, lung troubles. SBT leaves have been reported to possess several pharmacological properties including immunomodulatory, antioxidant, anti-inflammatory, antimicrobial and tissue regeneration etc. The present study was undertaken to evaluate the adjuvant property of supercritical carbon dioxide extracts (SCEs 300ET and 350ET) of SBT leaves in balb/c mice immunized with Tetanus and Diphtheria toxoids. The dynamic changes in the immune response were measured in terms of humoral and cell-mediated immune responses. We have seen the effect of SCEs on immunoglobulin subtypes and secondary immune response generation. In addition, the effect of SCEs on antigen specific cellular immunity was evaluated. Our results show that SCEs 300ET and 350ET significantly enhanced antibody titers in response to both TT and DT antigens. The secondary immune response generated was significantly increased in case of TT immunized animals. SCEs also enhanced cytokine levels (IFN-γ, IL-4, TNF-α and IL-1ß) and increased lymphoproliferation. Besides, both SCEs did not show any toxic effects. Therefore, the study suggests that SCEs are safe and have potent immunostimulatory activity and hence, seems to be a promising balanced Th1 and Th2 directing immunological adjuvant for various veterinary as well as human vaccines.

Curcumin Monoglucoside Shows Improved Bioavailability and Mitigates Rotenone Induced Neurotoxicity in Cell and Drosophila Models of Parkinson's Disease.

Curcumin (CUR), a dietary polyphenol has diverse pharmacologic effects, but is limited by poor bioavailability. This is probably due to decreased solubility, cellular uptake and stability. In order to enhance its solubility and bioavailability, we synthesized the CUR bioconjugate curcumin monoglucoside (CMG) and tested its bioavailability, neuroprotective and anti-apoptotic propensity against rotenone (ROT) induced toxicity in N27 dopaminergic neuronal cells and Drosophila models. Our results elucidate that CMG showed improved bioavailability than CUR in N27 cells. Pre-treatment with CMG protected against ROT neurotoxicity and exerted antioxidant effects by replenishing cellular glutathione levels and significantly decreasing reactive species. CMG pre-treatment also restored mitochondrial complex I and IV activities inhibited by ROT. ROT-induced nuclear damage was also restored by CMG as confirmed by comet assay. CMG induced anti-apoptotic effects was substantiated by decreased phosporylation of JNK3 and c-jun, which in turn decreased the cleavage of pro-caspase 3. Q-PCR analysis of redox genes showed up-regulation of NOS2 and down-regulation of NQO1 upon ROT exposure and this was attenuated by CMG pre-treatment. Studies in the Drosophila ROT model revealed that, CMG administration showed better survival rate and locomotor activity, improved antioxidant activity and dopamine content than ROT treated group and was comparable with the CUR group. Based on these data, we surmise that CMG has improved bioavailability and offered neuroprotection comparable with CUR, against ROT-induced toxicity both in dopaminergic neuronal cell line and Drosophila models, with therapeutic implications for PD.

Cardiology--a saga of one hundred years "but, where is the conductor?"--Braunwald, 2003.

Modern Cardiology was born with the birth of ECG invented by Einthoven in 1903. Since then a galaxy of great pioneers in succession led Cardiology through unprecedented landmark achievements to the present state which is as promising as challenging. This article reviews the journey of Cardiology as a speciality in the last one hundred years of its noteworthy life while highlighting the future directions and challenges the specialty is likely to face."

Lymphatic filariasis of the ovary and mesosalpinx.

We report 2 cases of filariasis, one in the ovary and the other in the mesosalpinx. In the first case, the patient underwent panhystrectomy and in the second case, right ovarian cystectomy with right salpingectomy were performed under general anaesthesia. Histopathology showed adult filarial worms in the dilated lymphatics of the right ovary, in the first case and in the mesosalpinx, in the second case. Both patients presented with complaints related to gynecological problems and not filariasis. Reports of filariasis in the literature and possible treatments and prevention strategies are also discussed.

Histopathology of Trypanosoma (Trypanozoon) evansi infection in bandicoot rat. I. visceral organs.

Experimental infection of Trypanosoma (Trypanozoon) evansi in Bandicota bengalensis produces an acute disease course leading to untimely death of the bandicoot rat. The sequential alteration of liver, spleen, lung, kidney, and heart was studied on the 5th, 8th, 12th, and 14th days postinoculation. The rats showed inflammatory, degenerative, and necrotic changes in these organs. In liver, pseudolobule formation, necrosis and hemorrhage within the sinusoids, and fatty degeneration of hepatic cells were the predominant histopathological changes. The changes were destructive and irreversible. In spleen giant cells aggregation and granulomatous lesion, i.e., accumulation of histiocytes, were the protective changes, whereas tissue and cell damage indicated irreversible degeneration. The gradual development of intrabronchus inflammation, aggregation of inflammatory cells around the alveoli, congestion of bronchioles, septal edema, atrophy of alveolar walls, migration of macrophages, and emphysema were the histopathological changes noticed in the lungs of the infected rats. The affected kidney showed infiltration of lymphocytes, hemorrhage in the interlobular space, and glomerulitis as the irreversible and destructive changes in the rats. There was degeneration of myocardium in the hearts of the rats. The histopathological changes in these organs are compared with those studied in surra, human sleeping sickness disease, and African trypanosomiasis. Possible mechanisms for these histological changes in the visceral organs are discussed.

Synthesis of terminally labelled RNA sequences: fluorescence and hybridisation study of RNA-DNA duplexes.

A stable and suitably labelled DNA-RNA hybrid, resistant to RNAse degradation has been prepared by hybridising the RNA sequence carrying methoxy group at 2'-position of ribose moiety in adenosines and labelled with dansyl group attached via a linker at the terminal uridyl unit. Study may be useful in sequencing and targeting specific viral sequences.

Study on curcumin-oligonucleotide conjugate as a probable anticancer agent: its hybridisation with telomere target sequence 5'-GGGATTGGGATT-3'.

Curcumin-oligonucleotide conjugate was synthesised by attaching diglycyl conjugate of curcumin to 12-mer complementary telomere sequence 5'-AATCCCAATCCC-3'. An enhanced Tm of 6 degrees C was found, showing high affinity for the target strand. This may be exploited for the suppression of cancer i.e. by blocking the expression of telomere sequence [GGGATT]n repeats.

Thermal endometrial ablation: a simple technique.

BACKGROUND: To evaluate the effectiveness and safety of a technique for thermal endometrial ablation. METHOD: The study was carried out in three steps; step I on 10 fresh uterine specimens, step II on 14 intact uteri during hysterectomy and step III in five patients, two weeks prior to hysterectomy. A simple device comprising a Foley catheter No. 14F, a three way cannula and a 20 cc syringe was used for the procedure. The balloon of the catheter was inflated within the uterine cavity with boiling normal saline for 9 minutes. Uterine cavity and serosal temperature was recorded during the procedure. Thermal injury was assessed by gross and histological examination of the specimens. RESULTS: The mean maximum endometrial cavity temperature was 91 degrees C whereas serosal temperatures in steps I and II were 34 degrees C and 35.9 degrees C respectively. None of the balloons ruptured in any of the steps and there were no complications. In 13 of 14 specimens in step II, there was a zone of hyperemia with a depth of 3 to 7 mm. The corresponding histological picture was extensive hemorrhage and fragmentation of glands throughout the endometrium including the cornual regions. Consistent findings were noted in step III in which all but one showed extensive coagulative necrosis of the whole of endometrium with edema of the underlying myometrium. There was a statistically significant positive correlation between the volume of fluid injected and depth of hyperemia. CONCLUSION: This technique could be a simple, effective, inexpensive and safe alternative to hysteroscopic endometrial ablation.

Synthesis and evaluation of novel bioconjugates as antiviral agents.

Novel lipid (mannito-stearate) antiviral nucleoside and oligonucleotide conjugates were prepared with improved lipophilic and membrane associating properties of drugs. Potential advantages of these liponucleotide prodrugs are lower toxicity, increased cellular uptake, nuclease resistivity and antiviral activity. Oligonucleotide conjugate complementary to a unique segment of viral genome may selectively disrupt the processes dependent on the segment by hybridisation.

Comparison of magnified chemiluminescent examination with incandescent light examination and colposcopy for detection of cervical neoplasia.

Magnified Chemiluminescent Examination (MCE) or speculoscopy is a new visual method for detection of cervical neoplasia. It utilizes low magnification and a special "blue-white" chemiluminescent light. The study includes 125 women with unhealthy cervix who were subjected to magnified examination with projected incandescent light (PIL) and chemiluminescent light (MCE). This was followed by colposcopy and directed biopsy from acetowhite areas. Of all 125 subjects, 20 patients showed cervical neoplasia of varying degree on histopathology. MCE could detect 18/20 neoplasias while PIL detected 11/20 cases. Sensitivity of MCE (90%) was significantly superior (P < 0.05) to PIL (55%) in detecting cervical neoplasia. Colposcopy, as compared to MCE, is better than speculoscopy as it facilitates grading of lesions due to higher magnification but antecedent MCE detected the acetowhitening in majority of patients (32/43). However, tendency to "overall" acetowhite lesions was significantly less during MCE when compared with colposcopy. Thus, MCE is a new diagnostic tool which is better than routine incandescent light examination and correlates well with colposcopy.

Early biologic behavior of bone grafts. A fine needle aspiration cytology study.

OBJECTIVE: To conceptualize, with fine needle aspiration cytology (FNAC), the early cellular events occurring in and around fresh autogenous and allogenic bone grafts during the first 40 postimplantation days. STUDY DESIGN: Forty-eight cases of bone grafts were studied by FNAC at serial intervals of 10, 20, 30 and 40 postimplantation days. Twenty patients were recipients of autogenous grafts, 16 received 0.6N HCI partially decalcified allogenic bone implants, and 4 received combined autogenous and allogenic bone grafts (included in the allograft group). There were eight control cases of closed fracture shaft femur, which were managed conservatively. RESULTS: The initial cellular responses in autogenous grafts, allografts and controls appear to be a part of the nonspecific reparative process followed by a more specific phase, with a steady increase in relative lymphocyte count from the 20th day onwards. Osteogenesis, as judged by osteoblasts and osteoclasts, was also comparable. CONCLUSION: Partially decalcified allografts appear to be a good substitute for autogenous bone grafts in clinical practice when adequate autogenous material is not available. FNAC is a good technique for studying bone graft responses without interfering with graft uptake. It is helpful in the early detection of subclinical infection or any other pathology at the graft site.

Comparison of pleural fluid cytology and pleural biopsy in the evaluation of pleural effusion.

Pleural fluid cytology and pleural biopsy results were studied in 65 cases of pleural effusion. The efficacy of pleural biopsy in diagnosis of neoplastic and non-neoplastic pleural diseases was compared. Of the 24 cases with confirmatory evidence of cancer, 17(70.8%) has positive cytologic findings in pleural fluid, whereas pleural biopsy was diagnostic in only 13 cases (54.1%). For non-malignant pleural effusion in 41 cases 40(97.5%) has a definite diagnosis (tuberculous pleuritis, acute fibrinous pleuritis or hydatid cyst) which could be made by cytology while only 31(75.6%) out of 41 were diagnosed on pleural biopsy. The study indicates that cytologic evaluation of pleural fluid is more efficaceous in the diagnosis of malignant and non-malignant pleural disease than percutaneous pleural biopsy.

Glyoxalase III from Escherichia coli: a single novel enzyme for the conversion of methylglyoxal into D-lactate without reduced glutathione.

A single novel enzyme, glyoxalase III, which catalyses the conversion of methylglyoxal into D-lactate without involvement of GSH, has been detected in and purified from Escherichia coli. Of several carbonyl compounds tested, only the alpha-ketoaldehydes methylglyoxal and phenylglyoxal were found to be substrates for this enzyme. Glyoxalase III is active over a wide range of pH with no sharp pH optimum. In its native form it has an M(r) of 82000 +/- 2000, and it is composed of two subunits of equal M(r). Glutathione analogues, which are inhibitors of glyoxalase I, do not inhibit glyoxalase III. Glyoxalase III is found to be sensitive to thiol-blocking reagents. The p-hydroxymercuribenzoate-inactivated enzyme could be almost completely re-activated by dithiothreitol and other thiol-group-containing compounds, indicating the possible involvement of thiol group(s) at or near the active site of the enzyme.

Purification and characterization of adipocyte heparan sulfate proteoglycans with affinity for lipoprotein lipase.

Lipoprotein lipase (LPL)-binding heparan sulfate proteoglycans (HSPGs) were isolated from cell extracts and conditioned media of cultured adipocytes treated with phosphatidylinositol-specific phospholipase C (PIPLC). The methodology employed included anion exchange chromatography, affinity chromatography on LPL Affi-Prep 10 and hydrophobic chromatography. HSPGs were resolved into two distinct fractions on the Octyl-Sepharose CL-4B matrix. Treatment of the eluted fractions with heparinase and heparitinase yielded core proteins of 48.4 and 39 kDa. The 39-kDa core protein is anchored to the cell surface by a glycosyl phosphatidylinositol anchor as evidenced by 1) release of the HSPG with the 39-kDa core protein into media by PIPLC treatment and 2) biosynthetic incorporation of [3H]ethanolamine and [32P]orthophosphate into the PIPLC-releasable 39-kDa core protein. PIPLC released 23% of the total heparin-releasable LPL. A similar percentage (24.5%) of the total heparan sulfate chains was released by PIPLC. Over 96% of the total adipocyte heparan sulfate chains bound to LPL Affi-Prep 10 column. The heterogeneity of core proteins of HSPGs with affinity for LPL may provide a structural basis for the multiple fates of LPL on the surface of adipocytes, i.e. internalization, degradation, or recycling to the cell surface and translocation into the medium.

Malignant ameloblastoma. A case report.

The cytologic features of malignant ameloblastoma, a rare tumor of the jaw, are described. Fine needle aspiration smears showed high cellularity, with small basaloid and polygonal epithelial cells with mild to moderate pleomorphism lying singly or in clusters. In Giemsa-stained smears, pinkish stroma with stellate cells or irregularly shaped nuclei and isolated spindle cells were seen in the background. Histopathology of the tumor showed sheets of cells with features of malignancylike atypical mitosis and vascular invasion. The tumor cells stained positive for keratin on immunohistochemistry. A review of cytology smears revealed that the features were sufficiently characteristic of an ameloblastoma, but it was difficult to label the lesion malignant on fine needle aspiration cytology.

Nucleolar organizer regions in FNAC smears of breast lesions.

The argyrophil technique for staining proteins associated with nucleolar organizing regions was applied to the fine-needle aspiration cytology smears of 45 cases of breast disease. These included 14 malignant and 31 benign lesions. A correlation with histology sections was done in 12 cases. There was a significant difference between AgNOR count of benign and malignant breast disease with one case out of 45 falling in the "gray" zone of overlap. Infiltrating lobular carcinoma was found to have the lowest NOR counts among the malignant lesions. The smear staining was superior to that on sections, the positive features being lack of background staining and better dispersion of NORs in the nucleoli.

Fine needle aspiration cytology of a lipoblastoma.

A lipoblastoma, an uncommon tumor of childhood that can be mistaken for a liposarcoma, was preoperatively diagnosed by fine needle aspiration cytology. The characteristic features on the cytologic smears were the presence of immature fat cells in the form of spindle-shaped cells, stellate cells and vacuolated lipoblasts along with lipocytes. The cytologic diagnosis was confirmed by histologic study of the excised tumor.

Fine needle aspiration cytodiagnosis of subcutaneous sacrococcygeal myxopapillary ependymoma. A case report.

A tumor located subcutaneously over the coccyx of a 32-year-old woman was diagnosed by fine needle aspiration (FNA) cytology as a myxopapillary ependymoma originating in the soft tissue. The FNA smears showed characteristic papillary structures, consisting of a central mucinous core surrounded by cuboidal-to-columnar cells, and scattered ependymal cells. The excised tumor was connected to neither the filum terminale nor the cauda equina. Histopathologic study confirmed the cytologic findings.

Use of homologous platelet factors in achieving total healing of recalcitrant skin ulcers.

We used homologous platelet-derived wound healing factors (HPDWHF) to achieve complete healing of recalcitrant ulcers of diverse cause. Twenty-three patients with 27 skin ulcers who had been receiving conventional wound care with no evidence of healing for an average period of 25 weeks (range, 12 to 156 weeks) were studied. The patients were first subjected to controlled wound care for 3 months, with saline solution and silver sulfadiazine dressings used in all cases. At the end of this period, persistent nonhealing ulcers were treated by topical use of HPDWHF and silver sulfadiazine. Ulcer parameters were recorded on the first day and every week during therapy until complete epithelization was achieved in either group. Each ulcer acted as its own control. In the controlled wound care group, only three ulcers in three patients achieved complete healing; the remaining 24 ulcers in 20 patients failed to achieve even 50% healing in the stipulated 3-month period. However, when subjected to HPDWHF applications, these ulcers healed completely, 100% healing occurring in 9.67 +/- 4.9 weeks (range, 3 to 19 weeks), which is highly significant (p less than 0.01). The healing response to HPDWHF applications was of uniform progression over the weeks. Only the basic cause of the ulcer determined the healing rates in this group. The shortest and the longest time to achieve 100% healing occurred in patients with diabetes (6.88 +/- 2.97 weeks) and in the venous stasis group (14.00 +/- 7.07 weeks). Age, sex, location of ulcer, ulcer duration, and ulcer measurements had no influence on the HPDWHF-stimulated healing rates. This is the earliest report of HPDWHF-stimulated repair in chronic nonhealing skin ulcers.