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[Altered behavioral response to centrally acting drugs in mice lacking PACAP].

Mice lacking PACAP (PACAP-KO) exhibits altered psychomotor behaviors, including impaired habituation to a novel environment and perseverative jumping, with a slightly reduced levels of the serotonin metabolite, 5-HIAA, in the brain. We have recently demonstrated that PACAP-KO exhibits abnormalities in sensorimotor gating as measured by prepulse inhibition (PPI) of the acoustic startle. In the present study, behavioral responses to centrally acting drugs (amphetamine, haloperidol, risperidone, fluoxetine, and 8-OH-DPAT) were examined in PACAP-KO. Surprisingly, a psychostimulant amphetamine effectively normalized the deficit in PPI as well as hyperactivity and jumping behavior. These results implied phenotypic and pharmacological similarity between PACAP-KO and attention deficit hyperactivity disorder (ADHD). Although a potent dopamine D2-like receptor antagonist, haloperidol, ameliorated the hyperactivity and jumping behavior, it had no effect on the deficit in PPI. In contrast, a prototype of serotonin-dopamine antagonist (SDA), risperidone, effectively normalized the deficit in PPI as well as hyperactivity, and jumping behavior. A selective serotonin reuptake inhibitor (SSRI), fluoxetine, also suppressed the hyperactivity and jumping behavior. A 5-HT1A receptor agonist, 8-OH-DPAT, significantly lowered rectal temperature in wild-type mice, while it had only a small effect in PACAP-KO. These results suggest the involvement of dopaminergic and serotonergic dysfunction in phenotypic changes observed in PACAP-KO.

Amphetamine↗

[The usefulness of tubular and glomerular proteinuria measurement in non-invasive diagnosis of vascular changes accompanying chronic allograft nephropathy].

Chronic allograft nephropathy (CAN) is the most important cause of late renal allograft loss. The standard diagnosis of CAN is based on pathological examinations according to Banff'97 scheme. The aim of the study was to evaluate the usefulness of tubular and glomerular proteinuria in non-invasive recognition of vascular changes accompanying CAN (AH--arteriolar hyaline thickening, CV--vascular fibrous intimal thickening). beta 2- and alpha 2-microglobulin (beta 2-m and alpha 2-m), albumin (alb), immunoglobulin G (IgG), total protein (tp) and creatinine (cr) concentration were measured in the second time urine specimen in 66 renal allograft recipients. Then the subsequent renal biopsies were done. The aim of statistical analysis (MANOVA, Stepwise Discriminant Analysis, SDA) was to diagnose CV and AH changes based on results of urine analysis listed above and the patient's age, time after transplantation and serum creatinine level (scr). Results obtained with statistical analysis were in 90.91% and 87.69% identical with CV and AH pathological diagnoses, respectively.

Adult↗

Efficacy of Euphorbia splendens and Leonotis nepetaefolia on aflatoxin producing fungi Aspergillus flavus and Aspergillus parasiticus.

Efficacy of three different concentrations (5, 10 and 15 mg/ml) of dry flower powder of E. splendens and L. nepetaefolia was tested on the growth of aflatoxin-producing toxigenic strains of fungi A. flavus (NCBT 101) and A. parasiticus (NCBT 128) in Sabouraud dextrose agar medium (SDA). Maximum (75%) inhibition of growth of A. flavus was seen at 15 mg/ml concentration of E. splendens flower dry powder, while A. parasiticus showed 50% inhibition of growth at 10 and 15 mg/ml concentrations. Total inhibition (100%) of growth of A. flavus was seen at 10 and 15 mg/ml for L. nepetaefolia and maximum (75%) inhibition of growth was seen for A. parasiticus at 15 mg/ml concentration. Bioassay with groundnut seeds soaked with different concentrations of flower extract proved that both fungi were incapable of infecting the seeds at 10 and 15 mg/ml of L. nepetaefolia flower extracts.

Aflatoxins↗

A comparison of a new affinity column system with a conventional tube LISS-antiglobulin test for antibody detection.

A recently introduced system for antibody detection (ReACT) consists of affinity columns (AFC) that contain protein A and protein G-coated agarose. We compared the ReACT system to a conventional tube low-ionic-strength saline antiglobulin test (LISS-AGT). We selected 100 LISS-AGT positive samples with clinically important and benign antibodies of varying strengths and 130 LISS-AGT negative samples to evaluate by the AFC method. AFC tests were positive with all 84 clinically important antibodies, including 36 antibodies that reacted <= 1+ at LISS-AGT (0% falsely negative). Eleven of 16 (69%) clinically benign antibodies reacted by AFC. Five samples (2 anti-Sda, 2 anti-I, and 1 inconclusive) were negative by AFC. AFC tests were negative with all 130 samples that were negative by LISS-AGT (0% falsely positive). The AFC method showed results comparable with results obtained with a conventional tube LISS-AGT for detection of clinically important antibodies. Some unwanted, clinically benign antibodies may not be detected by the AFC method.

Journal Article↗

Direct detection of Mycobacterium tuberculosis complex in pulmonary and extrapulmonary samples by BDProbeTec ET system.

BDProbeTec ET (Becton Dickinson, Sparks, Md, USA) is a fully automated walkaway system based on strand displacement amplification (SDA) technology that provides a method for the direct detection of Mycobacterium tuberculosis complex (MTBC) target sequence. The purpose of this study was to evaluate the ability of BDProbeTec ET system to detect MTBC directly from clinical specimens and compare the results with staining and culture. From February 2002 through December 2003 a total of 1521 [pulmonary (n=1329) and extrapulmonary (n=192)] specimens from 1518 patients were examined by BDProbeTec ET system for the detection of MTBC and the results were compared to those obtained by microscopy and liquid culture (BACTEC 9000 MB, Becton Dickinson). MTBC was cultivated from 65 specimens (60 pulmonary and 5 extrapulmonary) of which 43 (66.2%) (42 pulmonary and 1 extrapulmonary) were smear positive and 22 (33.8%) (18 pulmonary and 4 extrapulmonary) were smear negative. BDProbeTec ET detected MTBC in 58 (55 pulmonary and 3 extrapulmonary) of the 65 culture-positive specimens. Although the BDProbeTec ET system gave five false-negative results among the 18 smear-negative culture-positive pulmonary specimens, our results demonstrate that the BDProbeTec ET system is a reliable tool in smear-positive samples and given its technical characteristics it can be used for the rapid detection of MTBC in either pulmonary or extrapulmonary samples.

Bronchoalveolar Lavage Fluid↗

The P1H antigen and antibody.

P1H. a newly discovered compound antigen associated with both the ABO and P systems, occurs in approximately percent of Natal (South African) blacks. The compound antigen, is evident only when the red cells have exceptionally strong expression of both and H antigens, and it is apparently a dominant character. The antigen is thought to originate by steric rearrangement in the molecule, or to be the product of competition between pl and H gene transferases for the available paragloboside The corresponding antibody, anti-P1H, has been made by both P1 and P2 people and is a weak cold agglutinin It is not adsorbed by red cell, carrying either strongly expressed P1 or H antigens but is adsorbed by and eluted from P1H+ red cells sensitized by anti- P1H. The antibody is, inhibited by P1 but not by H. Lewis, or Sda substances.

Journal Article↗

[Dramatic increase in lymphogranuloma venereum among homosexual men in Hamburg].

Classical sexually transmitted diseases, including syphilis and gonorrhea, have recently increased significantly among homosexual men in Hamburg. During the last year we also observed an increase in patients with lymphogranuloma venereum (LGV) at the ifi-institute in Hamburg. In 2003, we identified 4 homosexual patients with LGV in different clinical stages. None of the patients has traveled outside Germany. Three of these patients were HIV-infected. In all cases Chlamydia trachomatis was identified by SDA (strand displacement amplification) in genital swabs or lymph node aspirates. In three cases sequencing of ompA PCR products was performed and in each instance revealed the C. trachomatis serovar L2. Other important genital pathogens were excluded by specific PCR tests, bacteriological and serological tests. LGV should be included in the differential diagnosis of anogenital and oral erosions, especially in homosexual patients and HIV-infected patients. Novel nucleic acid amplification tests can be used for the rapid and reliable diagnosis of LGV by identifying Chlamydia trachomatis.

Adult↗

[Expression of the human papillomavirus type 16L/E7 fusion protein in E. coli and observation of its immunogenicity in mice].

BACKGROUND: Many epidemiological and experimental evidences prove that cervical cancers are strongly associated with genital high-risk types of human papillomavirus (HPV). HPV16 is present in 50% of the tumor specimens. Thus, it is important to develop vaccines against HPV16 and cervical cancer. The authors studied the expression of the HPV16 L1DeltaCE7N fusion protein in E. coli and observed its immunogenicity. METHODS: The fragment of HPV16 L1DeltaC gene and the E7N gene were amplified by PCR separately; the fusion gene named L1DeltaCE7N was generated by fusing E7N to the C terminal of L1DeltaC then the chimeric gene was cloned into prokaryotic expression vector pGEX-2T and expressed in E. coli strain JM109. The L1DeltaCE7N protein expressed were detected by Western blot. Finally its immunogenicity was characterized in immunized mice. RESULTS: It was proved that the sequence and open reading frame of fusion gene L1DeltaE7N was correct by sequencing; SDA-PAGE gel analysis showed that HPV16 L1/E7 fusion protein was highly expressed in E. coli; the protein was expressed as soluble form and the molecular weight was about 85 x 10(3). The fusion protein could be purified by affinity chromatography and gel filtration. The ELISA result indicated that L1/E7 could elicit specific antibodies against L1 and E7 in immunized mice. In vivo tumor protection test indicated that tumor formation was retarded or prevented in the mice after vaccination with L1/E7, when C57 BL/6 mice were challenged by syngeneic HVP16E6 and E7 transformed tumor cells. CONCLUSION: HPV16L1/E7 fusion protein was expressed in E. coli, it can be a candidate for prophylactic and therapeutic vaccine for HPV16-associated infection and tumors.

Animals↗

Seromonitoring in small laboratory animal colonies. A five year survey: 1984-1988.

From 1984 to 1988 one thousand serologic investigations of laboratory animal colonies originating from 10 different European countries were performed. The most prevalent infections in mouse stocks were caused by Mouse hepatitis virus (MHV), Minute virus of mice (MVM), Theiler's encephalomyelitis virus (TEMV), Reovirus type 3 (Reo3), Sendaivirus, and Pneumonia virus of mice (PVM). In mice no infections with Lymphocytic choriomeningitis virus (LCM), Polyomavirus, Mouse adenovirus, and K-virus were recorded. Only two colonies were infected by Ectromelia virus. The first six virus infections of mice were also found in rat colonies as well as the rat-specific Coronaviruses (Sialodacryoadenitisvirus--SDA, Rat corona virus--RCV) and Parvovirus (Kilham rat virus--KRV, Toolan H-1 virus) being endemic. Antibodies to Bacillus piliformis were detectable in about 50% of rat stocks screened. This is in contrast to the mouse, where only about 10% of the colonies were found to be positive. A similar picture was seen for M. pulmonis which is primarily an infection of the rat. In mice no case was detected during the last two years. The number of investigations performed from guineapig, hamster and rabbit colonies was relatively low. Nevertheless, antibodies against the following antigens were detectable: In guineapig stocks: Reo3, PVM, Sendai, Simian virus 5 (SV5) and B. piliformis; in rabbits: Reo3, Sendavirus, SV5, and B. piliformis; in hamsters: PVM, LCM and B. piliformis. The overall contamination rate showed a continuous decrease until 1988. Nevertheless, about 50% of mouse and rat stocks still exhibited antibodies to one or more viral infections.

Animals↗

[Still's disease of the adult: analysis of a series of 9 cases].

Characteristic features of 9 patients (5 male, 4 female) affected by Still's disease of adult (SDA) were analysed. All of them had fever and in 7 cases it was associated to arthritis and cutaneous rash. The most frequently affected joints were knees, ankles, and carpus. The sedimentation rate was increased in all patients and 6 of them had associated leucocytosis. 5 patients had mild inflammation synovial fluid. The histological changes found were: cutaneous perivascular inflammation, reactive lymph node enlargement, sinusoid dilatation and hepatic periportal inflammation. The NSA were active in 4 patients but 5 needed steroids to control the systemic clinical alterations. 3 patients relapsed and one was diagnosed as having sarcoidosis 3 years later, the possible association not having being described. We concluded that suspicion is the most important factor for an early diagnosis.

Adolescent↗

Personal care attendants and severely disabled adults: attributions for relationship outcomes.

Although the personal care attendant (PCA) is identified as the single most important factor in maintaining the independence, health, and well-being of severely disabled adults (SDA), little research about the nature of the interpersonal relationship has been conducted. This study investigated the mutual perceptions of the SDAs and their PCAs of the outcome of their relationship and factors that promoted the outcome. Attribution theory was used to shape the study and guide analysis of the data. The majority of subjects reported mutually successful and satisfying relationships, and most attributed the outcome to their mutual ability, effort, or personal characteristics.

Activities of Daily Living↗

Revised calibration of the Reflotron cholesterol assay evaluated.

We evaluated the Boehringer Mannheim (B.M.) Reflotron Total Cholesterol "dry-chemistry" method after its recalibration in 1987. Reports in the literature up to 1986-1987 of a negative bias (up to -10%) in the method prompted a revision of the factory-set calibration of the Reflotron. For this, B.M. prepared a new set of calibrators with 12 different concentrations of cholesterol. We checked in two ways whether accuracy had been achieved: (a) The values assigned to the calibrators by B.M. were checked with the manual Abell-Kendall Reference Method (MAK) performed in an official Reference Center. These were shown to be correct. (b) Concurrently, a direct comparison was made by analyzing 200 fresh samples of human serum. Reflotron cholesterol values obtained for these samples proved to be accurate, meeting the current World Health Organization/Centers for Disease Control criterion of maximum bias less than or equal to 5%. Orthogonal regression analysis yielded the following correlation: Reflotron = 0.985 MAK + 0.238 mmol/L (y = ax + b). Reflotron mean = 6.26 mmol/L; MAK mean = 6.09 mmol/L. SDa = 0.015 mmol/L; SDb = 0.120 mmol/L, and r = 0.989.

Autoanalysis↗

Effect of spiced food on metabolic rate.

Since the time of Lavoisier it has been known that the ingestion of food in animals and man produces an increase in oxygen consumption. This increase in metabolic rate was originally called 'specific dynamic action' (SDA) and is now widely referred to as the thermic effect (TE) of food or diet-induced thermogenesis (DIT) (Rothwell & Stock, 1981). Much of the early work on the thermic effect was confined to the type and amount of food, notably the macronutrients--proteins, fats and carbohydrates. Later, it was shown that certain minor constituents of the diet such as caffeine and associated methylxanthines (Zahorska-Markrewicz, 1980; Jung et al., 1981) in tea and coffee could also have a profound effect on metabolic rate. The consumption of alcohol was also shown to increase metabolic rate (Rosenberg & Durnin, 1978). The work described in this paper reports the effect of another minor constituent of food, spices, on metabolic rate. Although the use of spices in our food has steadily increased with time little information exists on their effect on the metabolic rate. It has been estimated that approximately 40 different spices are used in our diet today. This communication reports the effect of chilli (red pepper, capsicum annuum) and mustard (Brassica juncea).

Adult↗

Identification of a novel ganglioside on erythrocytes with blood group Cad specificity.

The blood group Cad antigen is a carbohydrate structure well characterized on the sialoglycoproteins of the red cell membrane from some rare individuals (Blanchard, D., Cartron, J. P., Fournet, B., Montreuil, J., Van Halbeck, H., and Vliegenthart, J.F.G. (1983) J. Biol. Chem. 258, 7691-7695). However, protease treatment of whole cells did not destroy their antigenic activity which indicated that glycolipid might also be involved in the antigenic reaction. A crude ganglioside fraction was prepared from Cad cells and found to inhibit the hemagglutination reaction, whereas neutral glycolipids were inactive. Further analysis of the ganglioside extract from Cad erythrocytes by thin layer chromatography revealed an unusual profile characterized by a lower content of sialosylparagloboside and the presence of a novel ganglioside of slower mobility. Immunochemical studies demonstrate that this ganglioside binds Helix pomatia lectin and inhibits human anti-Sda antibody. In addition, a ganglioside with identical chromatographic mobility can be obtained by the enzymatic transfer of GalNAc from UDP-GalNAc to sialosylparagloboside using a microsomal preparation from human kidney. These results together with cell surface labeling experiments suggest that the major ganglioside of Cad erythrocytes might be derived from sialosylparagloboside by substitution with an additional N-acetylgalactosamine residue.

Animals↗

Isolation of a sperm membrane sialoglycoprotein autoantigen from rabbit testes.

A single rabbit sperm membrane autoantigen has been isolated from rabbit testes by immunoadsorbent chromatography, agarose chromatography, and electrophoresis. This rabbit sperm autoantigen (RSA-1) is located in the sperm plasma membrane and blocks the sperm cytotoxic (immobilizing) activity of autoantisera. RSA-1 is a sialoglycoprotein, approximately 40% carbohydrate and containing 0.2 micrograms sialic acid/microgram protein. Its m.w. is estimated at 13,000 +/- 1200 by SDA-PAGE. RSA-1 can self-aggregate into higher m.w. forms. Approximately 0.6 mg of RSA-1 can be isolated from 100 rabbits testes. The isolated RSA-1 shows a 300-fold increase in specific cytotoxic (immobilizing) inhibitory activity over the starting testis pellet material.

Animals↗

Structural and topological studies of cholesteatoma proteins, in relation to the keratinization process.

For the extraction of soft keratins from bovine snout or human epidermis the best results were obtained with a detergent (sodium dodecyl sulphate, SDS) or hydrogen bond breaking agent (urea) in a reducing medium (2-mercaptoethanol, 2-ME). N-acetylcysteine was a little less effective. Keratins from both sources gave typical sets of protein bands with molecular weights between 40.000 and 70.000 daltons. Upon electrofocusing special precautions had to be taken to avoid reoxidation and reaggregation of protein subunits. Keratins from aural cholesteatomas were obtained by extraction with SDA and 2-ME, with N-acetylcysteine alone and to a lesser extent with urea and 2-ME. The pattern of these keratins on SDS-gel is less complicated than that obtained from human skin or bovine snout. Histophotometric results argue for a clear analogy between the nuclear DNA metabolism in normal skin epidermis and cholesteatoma matrix. The only differences of potential relevance are the much wider range of the nuclear DNA amount in cholesteatoma stratum basale cells compared with basal cells in normal epidermis, and the higher persistence of nuclear DNA in cholesteatoma stratum granulosum, indicating postponement of keratinocyte terminal differentiation.

Animals↗

Primary structure of the oligosaccharide determinant of blood group Cad specificity.

Glycophorin A and B from Cad erythrocyte membranes are the carriers of the blood group Cad determinants. They are characterized by a significant increase in molecular mass, as compared to the corresponding glycophorins from control erythrocytes (Cartron, J.-P., and Blanchard, D. (1982) Biochem. J. 207, 497-504). Lipid-free glycophorin A, purified from Cad red cells, showed an increased GalNAc content in comparison to blood group B, Cad-negative, control cells. Alkaline-borohydride treatment of this Cad glycophorin A released as a predominant species a pentasaccharide; its structure was determined, by methylation analysis and by 500-MHz 1H-NMR studies, to be: (formula; see text) This novel oligosaccharide inhibited strongly the hemagglutination of Cad erythrocytes by the Dolichos biflorus lectin. It shares with the blood group Sda determinant a terminal GalNAc beta (1 leads to 4)Gal beta (1 leads to .) sequence.

Blood Group Antigens↗