Profiling expression patterns and isolating differentially expressed genes by cDNA microarray system with colorimetry detection

A high-density cDNA microarray with colorimetry detection system to simultaneously monitor the expression of many genes on nylon membrane is described and characterized. To quantify the expression of genes and to isolate differentially expressed genes, the southern hybridization process on filter membranes was employed. The levels of gene expression were represented by color intensities generated by colorimetric reactions in place of hazardous radioisotopes or costly laser-induced fluorescence detection. The gene expression patterns on nylon membranes were digitized by devices such as an economical flatbed scanner or a digital camera. The quantitative information of gene expression was retrieved by image analysis software. Quantitative comparison of the northern dot-blotting method with the microarray system is described. Applications employing single-color detection as well as dual-color detection to isolate differentially expressed genes among thousands of genes are demonstrated.     

Interleukin-6 production by rat granulosa cells in vitro: effects of cytokines, follicle-stimulating hormone, and cyclic 3',5'-adenosine monophosphate

In the present study we examined the influence of FSH as well as a number of well-established cytokines on interleukin (IL)-6 by rat granulosa cells in culture. Increasing concentrations of FSH, IL-1 alpha, IL-1 beta, tumor necrosis factor alpha (TNF alpha), and lipopolysaccharide (LPS) were incubated for 48 h with undifferentiated granulosa cells obtained from diethylstilbestrol-primed immature rats. The results demonstrate that FSH, IL-1 alpha, IL-1 beta, and LPS, but not TNF alpha, caused significant concentration-dependent increases in IL-6 release. We also examined the effects of dibutyryl-cAMP, forskolin, and 3-isobutyl-1-methyl-xanthine (IBMX) on IL-6 release by granulosa cells. Each of these agents caused a significant concentration-dependent increase in IL-6 production by granulosa cells in either the absence or presence of FSH. Taken together, these results show that the granulosa cell is not only a likely source of IL-6 but that the release of IL-6 can be regulated. Moreover, evidence suggests that cAMP may serve as a second messenger for the stimulated secretion of IL-6 by undifferentiated granulosa cells.     

Caffeine and other predictors of bone density among pre- and perimenopausal women

Cryopreservation has proved to be a highly successful method for long-term storage of viable embryos. The objective of this study on rat blastocysts was to define conditions for their cryopreservation. Three cryoprotectants, dimethyl sulfoxide, glycerol, and propanediol/sucrose, were compared in two cooling programs (to -30 or -80 degrees C) and two thawing protocols. The cooling was followed by immersion in liquid nitrogen. Programmed thawing was at the rate of 8 degrees C per minute; fast thawing consisted of direct exposure of the frozen embryos to the ambient laboratory temperature. The survival after the freeze/thaw was assessed from the post-thaw embryo morphology and ability to develop into apparently normal offspring in uteri of foster mothers (embryonic survival). The best method for preservation of rat blastocysts proved to be programmed cooling to -80 degrees C followed by fast thawing with glycerol as cryoprotectant (embryonic survival of 28.1%). In all the experimental groups, the proportion of embryos with good to excellent preservation of morphology was high. With dimethyl sulfoxide, after programmed cooling to -80 degrees C, embryonic survival was 9.9% (programmed thawing) and 17.5% (fast thawing). No embryos survived after programmed cooling to -30 degrees C. However, when the cryoprotectant was propanediol/sucrose, no difference was observed between programmed cooling to -80 degrees C with either method of thawing and programmed cooling to -30 degrees C and fast thawing (12.3, 6.2, and 8.0%, respectively).     

1,25(OH)2-vitamin D3 mediated changes in mRNA for c-myc and 1,25(OH)2D3 receptor in HL-60 cells and related subclones

The aim of this study was to clarify the change of the gastric mucosa following inoculation with Helicobacter pylori (H. pylori). Two pairs of cynomolgus monkeys received either H. pylori of human origin (group A) or H. pylori of monkey origin (group B) by intragastric inoculation at a dose of 10(9) CFU. After inoculation, endoscopical observation and biopsies were done every 7 days for one month. The bacteria in the biopsy samples were cultured quantitatively. The content of intracellular PAS-AB positive substance was quantitatively analyzed with the image analyzing system. Results were as follows: 1) Before inoculation, the gastric mucosa was endoscopically normal and free from H. pylori. 2) The quantity of H. pylori varied from 0 to 10(3) CFU/0.1 g tissue in group A and from 10 to 10(6) CFU/0.1 g tissue in group B. 3) Severe erosion was seen in group B, while mild mucosal erythema and erosion were observed in group A. 4) There was a correlation between the quantity of H. pylori and histological activity (Rauws' gastritis score). 5) The PAS-AB positive substance in H. pylori-positive mucosa was less than that in H. pylori-negative mucosa. In conclusion, gastric mucosal changes of varying degrees were brought about after inoculation of H. pylori depending on the quantity of bacilli in the gastric tissue.     

Downregulation of protein kinase C levels leads to inhibition of GnRH-stimulated gonadotropin secretion from dispersed pituitary cells of goldfish

In spring 1992 the Austrian Parliament has enacted a new law dealing with reproductive medicine. The 'Act on medically assisted conception' contains provisions which totally prohibit particular methods of assisted fertilization (such as embryo or egg donation). The main goal of the act, however, is to regulate procreative medicine and to amend the Austrian General Civil Code: the parents-to-be are now considered to have the legal rights and responsibilities of biological parents in any case.     

Effects of relative band intensity on prediction of protein secondary structure from CD

Increasing the magnitude of a protein CD spectrum obviously increases the magnitude of each predicted secondary structure by the same amount. However, increasing the magnitude of the negative, long-wave-length portion of a protein CD spectrum usually has the opposite effect from increasing the positive, short-wave-length portion. Thus small distortions in the CD spectra of proteins at short wavelength can have a significant effect on the analysis for secondary structure. This measurement error and its effect on the analysis are systematically investigated for 16 proteins of known structure. The results demonstrate that a two-point calibration of a CD instrument is mandatory to avoid serious errors when estimating secondary structure from protein CD spectra.     

Risperidone vs. haloperidol on reaction time, manual dexterity, and motor learning in treatment-resistant schizophrenia patients

BACKGROUND: The present study compared the effects of risperidone vs. haloperidol on reaction time, manual dexterity, and two types of motor learning in a sample of treatment-resistant schizophrenia patients. METHODS: Fifty-six DSM-III-R diagnosed schizophrenia inpatients participated in a randomized, double-blind comparison of risperidone vs. haloperidol. Measures of reaction time, manual dexterity, motor sequence learning, and gross motor learning were administered at baseline, after 4 weeks of fixed-dose medication, and after 4 weeks of flexible-dose medication. RESULTS: The results indicated that patients receiving risperidone showed greater improvement in reaction time and manual dexterity than patients receiving haloperidol. After covarying symptom changes and movement disorder ratings, the results remained significant. The two treatment groups did not differ on either measure of motor learning. CONCLUSIONS: The differences in performance in reaction time and manual dexterity may be due to a specific beneficial effect of risperidone, as opposed to a general reduction in extrapyramidal symptom liability, compared to haloperidol.     

Cuticular Hydrocarbons of Termites of the British Virgin Islands

A survey of the termites (Isoptera) of 17 islands of the British Virgin Island (BVI) complex yielded eight taxa belonging to three families. The Kalotermitidae include Neotermes mona (Banks), Cryptotermes brevis (Walker), Procryptotermes corniceps (Snyder), and an undetermined species of Incisitermes, likely Incisitermes nr snyderi (Light) or I. incisus (Silvestri). The only rhinotermitid collected is an undetermined species of Heterotermes (Froggatt). Parvitermes wolcotti (Snyder), Nasutitermes costalis (Holmgren), and N. acajutlae (Holmgren) comprise the Termitidae. Cuticular hydrocarbon mixtures were characterized for each of the taxa. Blends of abundant hydrocarbons are species-specific and can be used to identify a given taxon without the diagnostic castes, soldiers, or imagoes, although the species of Incisitermes were not separable on the basis of cuticular hydrocarbons.