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PubMedDate not supplied

Microscopic studies of accessory canals in primary molar furcations.

The purpose of this study was to evaluate the incidence of accessory canals in the furcation region of human primary second molars. Forty freshly extracted teeth were radectomized and furcations were separated. The severed pulp chamber floors were decalcified and dehydrated. Paraffin embedding followed and cross-cut serial sections were taken from the specimens. Microscopic examination of each section followed. Sixteen out of twenty (80 percent) of the maxillary and fifteen out of twenty (75 percent) of the mandibular primary second molars demonstrated accessory foramina in the furcation area. 17.3 percent of the accessory foramina were found in the pulp chamber floor and 82. 7 percent were observed interradicular, close to the periodontal ligament. Thirty percent of the primary second molars demonstrated accessory canals, running from the pulp chamber to the periodontal ligament. Within the limitations of a microscopic investigation it is assumed that accessory furcation canals might be responsible for interradicular bone pathology in case of pulpal inflammation or necrosis.

Decalcification Technique↗

PubMedDate not supplied

Fungistatic and fungicidal activity of east African medicinal plants.

Extracts of the traditionally used medicinal plants Entada abyssinica (stem bark), Terminalia spinosa (young branches), Harrisonia abyssinica (roots), Ximenia caffra (roots), Azadirachta indica (stem bark), Zanha africana (stem bark) and Spilanthes mauritiana (roots and flowers) were investigated for fungistatic and fungicidal activity against Candida spp. and Aspergillus spp. by a microtitre serial dilution technique. Entada abyssinica, T. spinosa, X. caffra, A. indica, and Z. africana showed activity against various Candida species. The minimum inhibitory concentrations (MICs) ranged from 0.006 to > 8 mg ml-1 and the minimum fungicidal concentrations (MFCs) from 0.06 to > 8 mg ml-1. Extracts from S. mauritiana (both roots and flowers) exhibited no activity against Candida spp., but against Aspergillus spp., the MIC and MFC values ranged from 0.13 to 0.25 mg ml-1 and from 0.13 to 1 mg ml-1 respectively. It is concluded that the extracts contain compounds with high antifungal potency.

Africa, Eastern↗

PubMed2003

Expression of cyclooxygenase-2 parallels expression of interleukin-1beta, interleukin-6 and NF-kappaB in human colorectal cancer.

Elevated expression of cyclooxygenase-2 (COX-2), the inducible isoform of prostaglandin H synthase, has been found in several human cancers, including colorectal cancer (CRC). This appears as a rationale for the chemopreventive effects of non-steroidal anti-inflammatory drugs in CRC. However, the reason for COX-2 overexpression is not fully understood. In cell culture experiments, COX-2 can be induced by proinflammatory cytokines, such as interleukin (IL)-1beta and IL-6. A crucial step in this signalling pathway is thought to be activation of transcription factor NF-kappaB. Based on these findings, we hypothesized an association between COX-2 overexpression and expression of IL-1beta, IL-6 and the NF-kappaB subunit p65 in human CRC. To test the hypothesis, we performed immunohistochemistry for the respective antigens on colorectal cancer specimens, obtained by surgical resections from 21 patients with CRC. Immunohistochemical results were confirmed by examination of protein levels in tissue lysates and nuclear extracts using western blotting. Non-neoplastic tissue specimens resected well outside the tumour border served as controls. COX-2 expression was found to be markedly enhanced in the neoplastic epithelium compared with controls. This was paralleled by a significantly higher expression of IL-1beta, IL-6 and p65. Serial sections revealed consistent cellular colocalizations of respective antigens in the neoplastic epithelium. Statistically, a significant correlation between expression of COX-2 and IL-1beta, IL-6 and p65 was found. Comparable results were obtained for stromal cells like macrophages and myofibroblasts. Further examination of nuclear extracts from CRC-specimens by western blotting confirmed a higher content of p65 protein compared with non-neoplastic control tissues. Therefore, our study provides evidence for an association between expression of COX-2 and IL-1beta, IL-6 and p65 in human CRC. The results are consistent with the thesis that proinflammatory cytokines such as IL-1beta and IL-6 may be accountable for the overexpression of COX-2 in CRC. Finally, the study corroborates a role for NF-kappaB in the control of COX-2 gene transcription in CRC. Given an antiapoptotic role for COX-2 in tumour cells, inhibition of NF-kappaB may offer an important strategy to interfere with the development and progression of CRC.

Aged↗

PubMed2004

Improved cognitive performance in human volunteers following administration of guarana (Paullinia cupana) extract: comparison and interaction with Panax ginseng.

Extracts from the plant guarana (Paullinia cupana) feature as putatively stimulating ingredients in a number of foods, drinks and dietary/herbal supplements. To date, little research in humans has examined the potential psychoactive effects of these extracts. Extracts of Panax ginseng, which are often sold in combination with guarana, contain similar potentially active components, and have been shown to modulate cognitive performance. In this double-blind, counterbalanced, placebo-controlled study, the cognitive and mood effects of separate single doses of: 75 mg of a dried ethanolic extract of guarana (approx 12% caffeine), 200 mg of Panax ginseng (G115), and their combination (75 mg/200 mg), were assessed in 28 healthy young (18-24) participants. On each day of the study (separated by a 7-day washout), cognitive performance and subjective mood were assessed pre-dose and at 1, 2.5, 4 and 6 h post-dose using the Cognitive Drug Research computerised assessment battery, Serial subtraction tasks and Bond-Lader mood scales. In comparison to placebo, all three treatments resulted in improved task performance throughout the day. In the case of guarana, improvements were seen across 'attention' tasks (but with some evidence of reduced accuracy), and on a sentence verification task. While also increasing the speed of attention task performance, both ginseng and the ginseng/guarana combination also enhanced the speed of memory task performance, with little evidence of modulated accuracy. Guarana and the combination, and to a lesser extent ginseng, also led to significant improvements in serial subtraction task performance. These results provide the first demonstration in humans of the psychoactive effects of guarana, and confirmation of the psychoactive properties of ginseng. Given the low caffeine content (9 mg) of this dose of guarana extract, the effects are unlikely to be attributable to its caffeine content.

Adult↗

PubMed1999

Circulating endothelin and nitrate-nitrite relate to hemodynamic and metabolic variables in human septic shock.

Activation of the nitric oxide (NO) pathway over that of endothelin in the vessel wall, as judged from circulating endothelin and nitrate-nitrite (NN) levels, may partly account for the hypotension associated with vasodilation, diminished catecholamine sensitiveness and O2 extraction, and lactic acidemia in human septic shock. In a prospective study, 14 consecutive patients with septic shock and a pulmonary artery catheter in place were included. For 3 days after admission, serial measurements of hemodynamic variables and plasma levels of endothelin and NN were done. The patients had a hyperdynamic circulation. Except for a higher final blood lactate level and more treatment with vasoconstricting catecholamines in nonsurvivors, global hemodynamic and O2-related variables did not differ between outcome groups. On the day of admission, circulating endothelin and NN levels were elevated and related to elevated levels of tumor necrosis factor-alpha and interleukin-6. The levels of endothelin increased in time in nonsurvivors as compared with survivors. The NN levels declined in survivors but not in nonsurvivors. The systemic vascular resistance indices (SVRI), global O2 extraction ratios, and blood lactate levels directly related to the endothelin levels. SVRI and global O2 extraction ratios inversely, and the lactate blood levels directly, related to NN levels, and the hemodynamic and metabolic parameters related directly to the ratio between endothelin and NN plasma levels on the days of the study. The vessel wall factors did not relate to the creatinine levels. The results suggest that the hemodynamic and metabolic peripheral abnormalities of human septic shock are mediated in part by cytokine-activated endothelin and NO systems in the vessel wall. They also suggest that increased production rather than diminished renal clearance accounts for elevated levels of NN and endothelin and that the latter are associated with a poor outcome.

Adult↗

PubMed1991

Treatment of hepatitis B virus-associated membranous nephropathy with adenine arabinoside and thymic extract.

Previously we found that corticosteroid treatment in the hepatitis B virus (HBV)-associated membranous nephropathy (HBVMN) was not associated with a favorable outcome. To distinguish the differences of the HBV DNA in macrophage, T and B cells among HBVMN patients with or without corticosteroid treatment, serial studies at different time points were investigated. HBV DNA appeared as an "episomal" molecule as with 3.2 kb in macrophage, T and B cells. This molecule disappeared after 12 months among HBVMN patients without corticosteroid treatment. HBV DNA, by contrast, appeared as episomal form even three years later in T cells, with frequent proteinuria among HBVMN patients with corticosteroid treatment. This finding indicates that the use of corticosteroids leads to a potential risk of enhancing HBV viral replication in T cells. We studied 24 HBVMN patients who had previously received corticosteroid treatment and had persistent proteinuria, who were administered combination therapy with adenine arabinoside for two weeks and thymic extract (Thymostimulin) for six months to decrease urine protein loss and obtain seroconversion. These 24 patients had heavy (22 of 24, 91.6%) or mild (2 of 24, 8.4%) proteinuria prior to adenine arabinoside and thymostimulin treatment. All 24 patients demonstrated HBV DNA in mononuclear cells and simultaneously exhibited sera positive with HBsAg and HBeAg. In contrast, after treatment only one case (4.2%) had heavy and two cases (8.4%) mild proteinuria; HBV DNA was demonstrated in macrophage (4 of 24, 16.7%), T cells (9 of 24, 37.5%), and B cells (6 of 24, 25%) as well as serum (24 of 24, 100%) prior to treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗

PubMed1999

Separation of sperm and vaginal cells based on ploidy, MHC class I-, CD45-, and cytokeratin expression for enhancement of DNA typing after sexual assault.

BACKGROUND: Successful DNA typing after rape is limited when only a few sperm and numerous vaginal cells are recovered from a swab, resulting in an extremely unfavorable ratio of male to female DNA. The goal of this study was to develop a protocol involving sperm cell sorting with flow cytometry based on differences in ploidy, major histocompatibility (MHC) class I, CD45 and cytokeratin expression. METHODS: Vaginal lavages were mixed with serially diluted ejaculate. After immunostaining and stoichiometric nuclear staining, spermatocytes were isolated by fluorescence-activated cell sorting. All sorted cells were used for DNA extraction and subsequent quantitative fluorescent multiplex polymerase chain reaction. The preferential lysis was performed for comparison. RESULTS: The sorting procedure was superior to the preferential lysis method within all tested dilutions. One documented case of rape was examined with both procedures and only after cell sorting with flow cytometry was the male DNA identified. CONCLUSIONS: We were able to show that separation of sperm and vaginal cells using cell sorting with flow cytometry may be crucial when there is only a few sperm detectable after rape.

Cell Separation↗

PubMed2006

Computer-based three-dimensional reconstruction of the anatomy of Microhedyle remanei (Marcus, 1953), an interstitial acochlidian gastropod from Bermuda.

Phylogenetic analysis of the highly enigmatic and aberrant acochlidian opisthobranch gastropods is mainly hindered by the lack of reliable anatomical data. Due to their small size, marine interstitial acochlidian species are not suitable for anatomical investigations by dissecting. The tinyness and complexity of, e.g., opisthobranch central nervous and reproductive systems make data derived from older paraffin-based histology and interpretation by hand-based graphical reconstruction questionable. In a former study, a hermaphroditic member of the basal acochlidian genus Hedylopsis ("Hedylopsacea") has been examined in detail. The present study uses the gonochoric Microhedyle remanei (Microhedylidae) as a model organism of the other traditional major acochlidian subgroup, the "Microhedylacea." More than 20 specimens with up to 2 mm body length were extracted from coarse subtidal sand near Castle Roads, Bermuda Islands. Their central nervous, digestive, circulatory, excretory, and genital systems were reconstructed three-dimensionally from serial semithin histological sections using AMIRA software. The radula was analyzed by SEM. Our specimens closely resemble the original description of M. remanei (Marcus, 1953; as Unela) from Brazil; apparent differences regarding the number of visceral loop ganglia or details of male genitalia are assumed to be due to oversights by the former author or because of different ontogenetic stages. Microhedyle remanei differs from all congeners due to the lack (vs. presence) of eyes; further distinguishing features are discussed. In contrast to members of the hedylopsacean Hedylopsidae, Acochlidiidae, and Tantulidae, M. remanei shows a nervous system with numerous precerebral accessory "ganglia" that are not differentiated into cortex and medulla. While all Microhedylidae previously were thought to lack a heart, M. remanei shows a two-chambered heart, as is usual for opisthobranchs. The oocytes of M. remanei are yolky and large in relation to body size and suggest intracapsular larval development. A comparative microanatomical discussion and the distribution of characters within acochlidian taxa indicate that the current classification of the Acochlidia does not reflect phylogenetic relationships. Detailed structural investigations on further poorly known species are required; the computer-based 3D reconstruction of semithin serial sections with AMIRA is shown to be an ideal tool for efficient analysis and presentation of the microanatomy of small specimens.

Animals↗

PubMed2005

Characterization of monoclonal antibody to L-homocysteic acid and its immunohistochemistry in rat hippocampus.

L-homocysteic acid (HCA) and other amino acids were conjugated to rat brain material (extracted rat brain protein) with glutaraldehyde to form HCA- and amino acids-brain material conjugates. The specificity of monoclonal antibody (McAb) was tested on serial dilution test and absorption test on enzyme-linked immunosorbent assay (ELISA) using these conjugates as antigens instead of amino acids-BSA (bovine serum albumin) conjugates used previously. The characterized McAb was applied for immunohistochemical staining using PAP (peroxidase antiperoxidase) technique in combination with silver enhancement of diamino-benzene (DAB) products. The results indicated that McAb to L-HCA reacted with L-HCA-brain material conjugates, but not with other amino acids-brain material conjugates so far tested. McAb absorbed with L-HCA-brain material abolished or decreased immunoreactivity of L-HCA-brain material with McAb. The antibody selectively stained subpopulation of cells and processes in the hippocampus fixed with glutaradehyde. Absorption of McAb with L-HCA-brain material abolished immunohistochemical staining. These results suggested that McAb was specific for L-HCA-brain materials and could be used for imuno-histocytochemistry. This would provide a new tool for immunohistochemical visualization and localization of L-HCA in the nervous system.

Animals↗

PubMed1982

Protein turnover in brain during the development of alcohol dependence.

The chronic effect of ethanol on central nervous system protein turnover was investigated in selected regions of brain following intoxication and withdrawal in a strain of ethanol preferring mice. Mice were serially injected with [14C]glucose in order to achieve a constant specific radioactivity of brain glutamate. Protein turnover was calculated from the specific activities of extracted protein and free glutamate. Results from these studies show that ethanol causes a significant increase in protein turnover in all sections of brain. The brain protein turnover in animals following alcohol withdrawal also shows an increase which deviates significantly from controls in 2 of the 3 regions examined.

Alcohol Drinking↗

PubMed2005

Immunoblot analysis of specific antigen bands predictable for Dirofilaria immitis infection in cats.

Serial sera from four mongrel cats experimentally inoculated with infectious larvae of Dirofilaria immitis were analyzed by immunoblot patterns against a phosphate buffered saline-extract of D. immitis. Antigen-specific protein bands detected indicate that the low molecular weight bands of 36, 32, 22, 19 and 14 kDa, are predictable for positive adult worm infection, suggesting diagnostic usefulness for adult D. immitis infection in cats.

Animals↗

PubMed1999

Separation of sperm and vaginal cells with flow cytometry for DNA typing after sexual assault.

BACKGROUND: Successful DNA typing after rape is limited when only a few sperm and numerous vaginal cells are recovered from a swab, resulting in an extremely unfavorable ratio of male to female DNA. The goal of this study was to develop a protocol involving sperm cell sorting with flow cytometry based on differences in ploidy, major histocompatibility class I, CD45, and cytokeratin expression. METHOD: Vaginal lavages were mixed with serially diluted ejaculate. After immunostaining and stoichiometric nuclear staining, spermatocytes were isolated by fluorescence-activated cell sorting. All sorted cells were used for DNA extraction and subsequent quantitative fluorescent multiplex polymerase chain reaction. The preferential lysis was performed for comparison. EXPERIENCE: The sorting procedure was superior to the preferential lysis method within all tested conditions. In unfavorable dilutions, the male DNA could be identified only after cell sorting with flow cytometry. CONCLUSION: We were able to show that separation of sperm and vaginal cells using cell sorting with flow cytometry may be crucial when there are only microtraces of sperm detectable after rape.

Adult↗

PubMed1989

Application of a new radiometric high-performance liquid chromatographic assay to define physostigmine pharmacokinetics in guinea pigs.

A sensitive high-performance liquid chromatographic method was developed to determine pharmacokinetic parameters of [3H]physostigmine from serial plasma samples from guinea pigs. Physostigmine was totally resolved from its metabolite, eseroline. The limit of sensitivity was 0.05 ng/ml from 0.2 ml plasma. Extraction efficiency was 99.6%. Within-run and among-run coefficients of variation (n = 6) for 0.2, 0.75, 1.5 and 2.5 ng/ml [3H]physostigmine ranged from 0.7 to 20% and 16 to 32%, respectively. Physostigmine (5 micrograms/kg) intramuscularly administered to the guinea pig (n = 6) reached maximum serum concentration (1.5 ng/ml) in 26 min. The apparent volume of distribution and systemic clearance were 1.4 l/kg and 26 ml/min/kg, respectively. This method was successful in defining physostigmine pharmacokinetic parameters in guinea pigs and can be employed for other small animal pharmacokinetic studies.

Animals↗

PubMed1999

Differential distribution of gonadotropin-releasing hormone variants in the brain of Hydrochaeris hydrochaeris (Mammalia, Rodentia).

1. In a previous paper we reported evidence for the presence of mGnRH- and sGnRH-like peptides in the preoptic-hypothalamic region of the capybara Hydrochaeris hydrochaeris (Montaner et al., 1998). In that study, the presence of a cGnRH-II like molecule in olfactory bulb extracts was suggested. 2. The capybara, the largest living rodent in the world, belongs to the order Hystricomorpha, which is considered to be one of the oldest groups of rodents. Some authors consider that this group is the ancestor of all remaining rodents. 3. In this study we have characterized GnRH molecular variants found in extracts from the olfactory bulbs and the mesencephalic region of capybara. These regions represent the two GnRH neuronal systems: the terminal nerve-septopreoptic and the midbrain systems. 4. An indirect method combining reverse-phase high-performance liquid chromatography (RP-HPLC) and radioimmunoassay (RIA) was used to characterize GnRH variants. The analysis of both extracts with two different RIA systems revealed three immunoreactive GnRH peaks, coeluting with mGnRH, cIIGnRH, and sGnRH synthetic standards. These results were additionally supported by serial dilution studies with specific antisera. 5. To our knowledge this the first report on the presence of three GnRH variants in the brain of an eutherian mammal. These results suggest that, similarly to other vertebrates, the expression of multiple GnRH variants may also be a common pattern in mammals.

Animals↗

PubMed2001

Improved detection of viral RNA isolated from liquid-based cytology samples.

BACKGROUND: Molecular diagnosis requires the ability to obtain high-quality nucleic acids that are representative of the disease state. We evaluated the recovery and detection of limiting amounts of viral oncogenic RNA from cells fixed in liquid-based cytology media. METHODS AND RESULTS: Serial dilutions of a human papillomavirus (HPV)-positive cell line fixed in a liquid media was used as a model system. Total nucleic acid (TNA) extraction produced RNA with clearly visible ribosomal bands even after one year of storage. These TNA extracts, treated with DNase-I, were used in an RT-PCR assay for HPV-16 E6-E7 oncogenic transcripts. With chemiluminscent Southern blot detection, samples with one HPV-positive cell in 30,000 were consistently detected. CONCLUSION: PreservCyt-fixed cells can yield RNA suitable for molecular assays even after one year of storage.

Blotting, Southern↗

PubMed1987

Biochemical changes in desmosomes of bovine muzzle epidermis during differentiation.

Biochemical changes taking place in desmosomes during differentiation have been studied. Bovine muzzle epidermis was sliced horizontally into 6 layers, 0.2 mm thick, and desmosomes were isolated from each layer. These were then analyzed by polyacrylamide gel electrophoresis. The electrophoretic patterns of desmosomal proteins from the 6 layers were found to be qualitatively similar to each other, but there was an increase in the ratio of the amount of 150 kD glycoprotein (desmoglein I) relative to 240 and 210 kD proteins (desmoplakins) in the upper layers of the epidermis. This finding was supported by the similar increase observed in electrophoretic patterns of proteins extracted directly from each layer of the epidermis in electrophoretic sample buffer. In order to study the fate of desmosomal components in the stratum corneum, serial skin surface biopsies were stained with antisera against desmosomal components using indirect immunofluorescence techniques. This experiment showed that desmosomal proteins and glycoproteins persist in the stratum corneum but quantitatively decrease in the outer layers. This decrease may play a significant role in desquamation.

Animals↗

PubMed1994

Skin elicitation threshold of ethylbutyl thiourea and mercaptobenzothiazole with relative leaching from sensitizing products.

We studied 3 contact sensitizers present in rubber products, ethylbutyl thiourea (EBT), 2-mercaptobenzothiazole (MBT) and 2,2-dithio-bis-benzothiazole (MBTS), to relate the amount of sensitizer eliciting allergic contact dermatitis to the quantity leaching from a product into various biological fluids: normal saline, human plasma and 3 synthetic sweat solutions of pH 5.5 to 7.5. To determine the amount of sensitizer remaining after leaching, Soxhlet extraction with acetonitrile was subsequently performed. High-performance liquid chromatography was used for chemical analysis. 12 MBT-sensitive patients were patch tested with serial dilutions of MBT and MBTS in petrolatum. A Latin Square design was used in statistical analysis of variance of the patch test results. Large amounts of thioureas leached from 2 rubber articles eliciting thiourea dermatitis, the literature suggesting that these would have been well above the elicitation threshold. Leaching of MBTS was relatively greater than MBT into most media, whereas MBT was a more potent elicitor than MBTS at equivalent concentrations. The lowest eliciting concentration of MBT in 1 subject was 0.01%. Such information should prove helpful to manufacturers in designed products that do not release allergens sufficiently to cause reactions in consumers.

Adolescent↗

PubMed2004

Doxorubicin pharmacokinetics following a single-dose infusion to sulphur-crested cockatoos (Cacatua galerita).

OBJECTIVE: To determine the pharmacokinetics of doxorubicin in sulphur-crested cockatoos, so that its use in clinical studies in birds can be considered. DESIGN: A pharmacokinetic study of doxorubicin, following a single intravenous (i.v.) infusion over 20 min, was performed in four healthy sulphur-crested cockatoos (Cacatua galerita). PROCEDURE: Birds were anaesthetised and both jugular veins were cannulated, one for doxorubicin infusion and the other for blood collection. Doxorubicin hydrochloride (2 mg/kg) in normal saline was infused i.v. over 20 min at a constant rate. Serial blood samples were collected for 96 h after initiation of the infusion. Plasma doxorubicin concentrations were assayed using an HPLC method involving ethyl acetate extraction, reverse-phase chromatography and fluorescence detection. The limit of quantification was 20 ng/mL. Established non-parametric methods were used for the analysis of plasma doxorubicin data. RESULTS: During the infusion the mean +/- SD for the Cmax of doxorubicin was 4037 +/- 2577 ng/mL. Plasma concentrations declined biexponentially immediately after the infusion was ceased. There was considerable intersubject variability in all pharmacokinetic variables. The terminal (beta-phase) half-life was 41.4 +/- 18.5 min, the systemic clearance (CI) was 45.7 +/- 18.0 mL/min/kg, the mean residence time (MRT) was 4.8 +/- 1.4 min, and the volume of distribution at steady state (V(SS)) was 238 +/- 131 mL/kg. The extrapolated area under the curve (AUC(0-infinity)) was 950 +/- 677 ng/mL x h. The reduced metabolite, doxorubicinol, was detected in the plasma of all four parrots but could be quantified in only one bird with the profile suggesting formation rate-limited pharmacokinetics of doxorubicinol. CONCLUSIONS AND CLINICAL RELEVANCE: Doxorubicin infusion in sulphur-crested cockatoos produced mild, transient inappetence. The volume of distribution per kilogram and terminal half-life were considerably smaller, but the clearance per kilogram was similar to or larger than reported in the dog, rat and humans. Traces of doxorubicinol, a metabolite of doxorubicin, were detected in the plasma.

Animals↗