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PubMed1993

Efferent projections of the sexually dimorphic area of the gerbil hypothalamus: anterograde identification and retrograde verification in males and females.

Outputs of the sexually dimorphic area (SDA) of the gerbil hypothalamus were identified by injecting Phaseolus vulgaris-leucoagglutinin into the medial or lateral SDA (mSDA, lSDA) in males and females. They were verified by injecting Fluoro-Gold or rhodamine-labeled beads into over half the areas that contained labeled fibers. Both anterograde and retrograde tracing showed that the mSDA and lSDA project to many of the same sites but often to differing degrees. The mSDA projects more heavily than the lSDA to many of their forebrain targets including the ventral part of the lateral septal nucleus, the bed nucleus of the stria terminalis, the medial tuberal area, and the anteroventral periventricular, arcuate, ventromedial and ventral premammillary nuclei of the hypothalamus. The lSDA projects more heavily than the mSDA to many of their mid- and hindbrain targets including the caudal, ventrolateral part of the periaqueductal gray, the retrorubral field, the pedunculopontine tegmental nucleus, and the locus coeruleus. In many other areas of the brain, the projections of the mSDA and lSDA are similar in size. These areas include the substantia innominata, the vascular organ of the lamina terminalis, the anterior amygdala, the posterior hypothalamus, the reuniens and paraventricular nuclei of the thalamus, and the pontine periaqueductal gray lateral to the fourth ventricle. The SDA pars compacta (SDApc), a small cell group embedded in the mSDA of males, projects to many fewer areas than the surrounding mSDA. It was strongly labeled when retrograde tracers were injected into the encapsulated part of the bed nucleus of the stria terminalis, the anteroventral periventricular nucleus, or the mSDA. It was also labeled from the vascular organ of the lamina terminalis, the caudal part of the lateral bed nucleus of the stria terminalis, the lSDA, the area lateral to the mSDA, the arcuate nucleus, the ventral premammillary nucleus, and the ventrolateral part of the ventromedial nucleus of the hypothalamus. Nothing resembling an SDApc was identified during retrograde tracing in females.

Animals↗

PubMed2004

Multiple sample amplification and genotyping integrated on a single electronic microarray.

We report a novel method that allows simultaneous in situ amplification and then genotyping of single nucleotide polymorphism (SNP) for multiple samples on a single electronic microarray. The locus coding for one of the common inherited thrombosis risk factors, Factor V Leiden (FVL), was chosen as a model system for SNP analysis. This method combines strand displacement amplification (SDA) with electrophoretic movement and concentration of DNA on electronic microarrays to provide a single platform for DNA amplification and analysis. The method includes: electronic anchoring of allele-specific SDA amplifiable primers (APs) and a nonamplifiable primer (NAP) to different electrodes, electronic hybridization of genomic DNA from different samples to those primers, in situ amplification of target DNA, and genotyping of FVL. Compared to previous anchored SDA methods, the addition of a NAP improves detection signals by at least 20-fold. The sensitivity of this method is dependent on the amplification time. Using this method, nine different genomic DNA samples with known FVL genotypes were amplified and correctly genotyped on a single electronic microarray without any contamination between samples. The present method could streamline development of nucleic acid-based assays in applications of molecular diagnostic, point-of-care testing, and forensic detection, which often require the capability to analyze multiple samples efficiently.

Base Sequence↗

PubMed1996

Effects of androgens and estrogens on sexual differentiation of sex behavior, scent marking, and the sexually dimorphic area of the gerbil hypothalamus.

Mating, marking and the sexually dimorphic area (SDA) were studied in male gerbils that were castrated or sham-operated on the day of birth and in females given testosterone (T) propionate or oil 1 day later. Other females received dihydrotestosterone (DHT), R1881, estradiol benzoate (EB), R2858, DHT and EB, or DHT and R2858. Females and control males were castrated as adults. Then half of each group was given T and tested for marking and male sex behavior. They were still exposed to T at perfusion. The rest were given EB and progesterone and were tested for marking and lordosis. They were perfused 1 month after hormone treatment ended. Neonatal castration eliminated mounting, decreased T-induced marking and increased lordosis in males. Neonatal exposure to TP decreased lordosis and increased mounting, intromission patterns and T-induced marking in females. DHT and R1881 did not affect marking or mating when given alone. EB and R2858 decreased lordosis and increased mounting. DHT enhanced the former effect but not the latter. Only combined treatments increased T-induced marking. Total SDA volume was not affected by neonatal treatments, but "dark volume" was largest in control males. The SDA pars compacta (SDApc) was seldom seen in females unless they received steroid neonatally but was present in males that were castrated neonatally. SDApcs of neonatal castrates and steroid-treated females differed, though, from SDApcs of control males in that they were smaller in the absence of T and did not enlarge when exposed to T. Another cell group, the cmSDApc, was affected by T neonatally but was not seen consistently in either sex. SDApc volume correlated positively with T-induced marking and an index of male sex behavior. Both SDApc and cmSDApc volumes correlated negatively with lordosis. The correlations with marking and male sex behavior probably reflect constraints on sexual differentiation rather than causal relationships since these behaviors persist when the SDApc is gone. The effects of direct manipulations of the SDApc on lordosis have not been studied.

Androgens↗

PubMed2002

Identification of Escherichia coli O157:H7 and other enterohemorrhagic serotypes by EHEC- hlyA targeting, strand displacement amplification, and fluorescence polarization.

Human disease caused by enterohemorrhagic E. coli O157:H7 and other serotypes (EHEC) has been associated with bovine fecal contamination of food and the environment. The range of serotypes, low infectious dose, and numerous transmission vehicles for EHEC render development of detection methods for this pathogen complex. In this study, the hemolysin gene (EHEC- hly A) was targeted with oligonucleotides, and probe-target hybrids were amplified using strand displacement amplification (SDA). Amplicons were resolved in the complete reaction mix through changes in the fluorescence polarization (FP) of a fluorescein-labeled detector probe hybridized to the amplicons during amplification. Results combining EHEC- hly A, SDA, and FP were obtained within 35 min of reaction initiation. The test specificity was determined on EHEC strains representing 13 serotypes (49 isolates); and control uropathogenic, commensal, and other organisms (10 isolates). Statistical analysis of results indicated a sensitivity in the reaction vessel to 4.3 bacteria (95% confidence interval), and a specificity for EHEC (n=59) at 100% (P=5.11E-17; i.e. P<<0.05). Detection based on combining EHEC- hly A, SDA, and FP was compatible with water sources directly associated with human infection (drinking and recreational supplies), and bovine drinking trough water representing an environmental matrix linked to the maintenance of an EHEC animal reservoir.

Animals↗

PubMed1996

Influence of marrow on ultrasonic velocity and attenuation in bovine trabecular bone.

Measurements of ultrasonic velocity and specific differential attenuation (SDA) were obtained on 24 bovine trabecular bone specimens from the femoral condyles. The measurements were obtained using two pairs of ultrasonic transducers, one with a low nominal center frequency (500 kHz) and the other pair with a high nominal center frequency (1 MHz). The ultrasonic velocity and specific differential attenuation associated with the bone samples were determined both with and without marrow, i.e., replacing the marrow with water in the pores of the trabecular bone. Significant increases (2.1% and 2.9%) in the velocity of ultrasound were observed after removal of the marrow, for the low and high frequency transducer pairs, respectively. In contrast, significant decreases (-6.5% and -8.8%) in SDA were observed after removal of the marrow, for the low and high frequency transducer pairs, respectively. The bone densities (BD) of the samples were also determined using single photon absorptiometry (SPA). Correlations between ultrasonic parameters and bone densities for samples both with and without marrow were found to be similar. For example, for the 1 MHz transducer pair, the correlation between BD and velocity was r = 0. 86 with marrow, and r = 0.89 without marrow. This study also compared the results obtained using a contact (no water bath) technique and an insertion (with a water bath) technique of ultrasonic measurements. For the high frequency transducer pair, the correlation coefficients between the two methods were r = 0.99 and r = 0.93, for the velocity and specific differential attenuation, respectively. Similar results were found for the low frequency transducer pair as well. In addition, approximately equal correlations between BD and ultrasonic velocity and SDA were also found, indicating that contact and insertion measurements provide essentially equivalent information.

Absorptiometry, Photon↗

PubMed2004

Abnormal resonance behavior of the postural control loop in Parkinson's disease.

Human postural control of upright stance sporadically can show an oscillatory behavior. Based on previous work, we assessed whether an abnormal tendency for such oscillations might contribute to the motor impairments in patients with basal ganglia dysfunction such as Parkinson's disease (PD). We investigated postural control during unperturbed stance in normal control subjects and in PD patients off and under treatment, focusing on stabilogram diffusion analysis (SDA) of the foot center of pressure (COP) excursions and conventional measures of the sway amplitude and velocity. We found abnormal 1 Hz body sway oscillation in the SDA curves of PD patients that differed significantly from the body sway typically observed in control subjects during quiet stance. The 1 Hz body sway oscillation was associated with abnormally large and fast sway in the patients off treatment. Under treatment with levodopa, with 'deep brain stimulation' (subthalamic nucleus) and even more so with combined treatment, the oscillations in the SDA curves vanished and the sway became slower. The loss of oscillation and reduction of sway velocity were highly correlated with the improvements of patients' clinical motor assessment score. However, sway amplitude was not correlated with the patients' motor assessment score and patients reported clinical improvement under therapy even though sway amplitude increased on average. A simple feedback model of the postural control system with abnormally large internal noise could predict experimental measures both on and off treatment. The off treatment condition was consistent with a high motor gain in the feedback loop, and the on treatment condition with a reduced motor gain.

Basal Ganglia↗

PubMed2006

The use of classification methods for modeling the antioxidant activity of flavonoid compounds.

A study using two classification methods (SDA and SIMCA) was carried out in this work with the aim of investigating the relationship between the structure of flavonoid compounds and their free-radical-scavenging ability. In this work, we report the use of chemometric methods (SDA and SIMCA) able to select the most relevant variables (steric, electronic, and topological) responsible for this ability. The results obtained with the SDA and SIMCA methods agree perfectly with our previous model, in which we used other chemometric methods (PCA, HCA and KNN) and are also corroborated with experimental results from the literature. This is a strong indication of how reliable the selection of variables is.

Antioxidants↗

PubMed1990

Purification, characterization, and structural elucidation of the active moiety of the previously called "suppressor activating factor (SAF)".

Upon extensive purification of the serum-free supernatant produced by a mutant T cell line (6T-CEM), an immunosuppressive activity was found to reside in an oxidized product of spermine, spermine dialdehyde (SDA). The activity was purified to homogeneity from a serum-free supernatant by using gel filtration chromatography and reverse-phase C18 HPLC. Fast Atom Bombardment (FAB) mass spectral analysis revealed its MW to be 202 and Electron Impact (EI) analysis of the acetylated material identified the purified molecule to be spermine. In the presence of human or rodent plasma, spermine exhibited no immunosuppressive activity up to 2 mg/ml. However, when assayed in the presence of FCS, which contains polyamine oxidase (PAO), spermine is oxidized to its corresponding dialdehyde which is active at 0.1 microM/ml. We have previously described a high molecular weight suppressor activating factor (SAF) found in the serum-containing supernatant of the 6T-CEM cell line. Our preliminary biological data suggest that SDA is probably responsible for the immunosuppressive activities previously observed for the SAF. The strong affinity of SDA for proteins and thiocompounds may account for the apparent high MW previously reported for SAF.

Aldehydes↗

PubMed1987

The adrenal gland of stranded whales (Kogia breviceps and Mesoplodon europaeus): morphology, hormonal contents, and biosynthesis of corticosteroids.

Histochemical assays, hormonal quantitation, and steroid biosynthetic studies were carried out with adrenal glands obtained from four stranded whales of two different species (Kogia breviceps and Mesoplodon europaeus), and selected comparisons were made with the results of similar studies of adrenals from terrestrial mammals (man, beef, rat). Histochemical chemical assays of the whale glands for succinic dehydrogenase activity (SDA) showed an intense SDA-positive reaction in the peripheral cortex, and an SDA-negative central medulla, a pattern similar to that found in terrestrial mammals; the whale adrenals, however, demonstrated a markedly pseudolobulated appearance because of a festooned corticomedullary junction. On radioimmunoassay of preformed cortical steroid hormones, corticosterone (B) exceeded cortisol (F) levels by a factor of 3 in the whale adrenals and aldosterone (Aldo) concentrations were 20-100 times lower than in the terrestrial mammals studied. HPLC determinations of preformed medullary catecholamines showed that, contrary to the findings in the terrestrial mammals studied, norepinephrine predominated over epinephrine and the levels of dopamine were much higher in the whale adrenals. In vitro, surviving sections of whale adrenals elaborated B from endogenous substrates, but not F or Aldo. Incubations of subcellular fractions of the whale adrenals with 14C-labeled precursors resulted in the isolation of several steroid intermediates (pregnenolone, progesterone, deoxycorticosterone) as well as the glucocorticoid end-product B, but again without evidence of the formation of either F or Aldo. In keeping with studies in terrestrial mammals, the enzymatic reactions involved in the conversion of [14C]cholesterol to B occurred under aerobic conditions, required the presence of an exogenous NADPH-generating system, and had identical subcellular localization in the whale adrenals. The process of steroid biosynthesis thus appears generally similar in aquatic and terrestrial mammals. It is possible that some of the unusual findings in the whale adrenals studies here may be related to the profound stress of stranding experienced by these marine mammals.

Adrenal Cortex Hormones↗

PubMed1993

Morphological and morphometric measurements in colorectal mucosa of subjects at increased risk for colonic neoplasia.

Measurements of intermediate biomarkers have recently increased, attempting to provide useful information about cancer risk. We report morphological findings in rectal mucosal biopsies from patients at low risk and at high risk for colorectal cancer. Rectal biopsies were analyzed from fourteen Seventh-Day Adventist (SDA) subjects at low risk and from twenty-seven members of families with hereditary nonpolyposis colonic cancer (HNPCC) at higher risk. The following measurements were made on rectal crypts: length of crypts, numbers of cells, diameter of the surface, middle and base of the crypts and infiltration of inflammatory cells into the lamina propria. Findings indicated morphological differences in normal-appearing rectal mucosa of individuals in the HNPCC group compared with SDA subjects (P < 0.05). They included shorter crypts with fewer epithelial cells and increased cellular infiltration in the mucosa of HNPCC subjects compared with SDA subjects, suggesting minimal inflammation, and an early stage of crypt atrophy in the rectal mucosa of subjects at higher risk for colonic neoplasia.

Colonic Neoplasms↗

PubMed2004

Temporal variation in the specific dynamic action of juvenile New Zealand rock lobsters, Jasus edwardsii.

To enhance the on-growing of Jasus edwardsii in culture, it is important to understand the feeding physiology of juveniles. In crustaceans, there is a loss of energy and an increase in oxygen consumption (specific dynamic action or SDA) associated with feeding. The present research measured the SDA of juvenile J. edwardsii fed either in the morning or at night held at 15 degrees C. Closed box respirometry was used to measure oxygen consumption (MO(2)) and ammonia excretion in juvenile lobsters. Juveniles exhibited a nocturnal rhythm in both MO(2) and ammonia excretion. The factorial rise in MO(2) (1.58+/-0.03 times) for lobsters fed in the morning was significantly less than lobsters fed at night (1.80+/-0.01 times). Lobsters fed in the morning had a significantly shorter SDA (30+/-1.2 h) response compared to lobsters fed at night (36+/-1 h). Energy loss as a result of digestion was less for lobsters fed in the morning. Therefore, if juvenile J. edwardsii are fed in the morning, they could optimise the energy content of the meal and this could result in increased growth.

Ammonia↗

PubMed2005

Influence of food type on specific dynamic action of the Chinese skink Eumeces chinensis.

We used the Chinese skink (Eumeces chinensis) as an experimental model to study influence of food type on specific dynamic action (SDA) of feeding. Thirty-three adult males collected from a natural population were divided equally into three (one control and two experimental) groups. We starved all skinks at 30 degrees C for 3 days and then provided the experimental skinks with a single meal consisting of either mealworms or meat [the flesh of the bullfrog (Rana catesbeiana)]. Food ingested by skinks of the two experimental groups differed in lipid content and lean dry mass but not in total dry mass and energy. Defecation following feeding occurred slightly earlier in skinks ingesting mealworms (mean=41.7 h) than in those ingesting meat (mean=47.7 h), but the difference was not significant. Analyses of variance (ANOVAs) with repeated measures showed that temporal variation in oxygen consumption over 72 h after feeding was evident in the experimental skinks but not in the control ones. Oxygen consumption was higher in the experimental skinks than in the control ones during the time interval between 4.5 and 36 h after feeding. The peak metabolic rate was greater but occurred later in skinks ingesting meat than in those ingesting mealworms. The estimated amounts of oxygen consumed by mealworm-fed, meat-fed and unfed skinks at 30 degrees C over 72 h after feeding were 356.5, 393.8 and 295.2 mL, respectively. Our results provide a support for the previous prediction that SDA is affected by types of food ingested by animals as skinks ingesting mealworms and meat differed in the time to reach a peak metabolic rate, the level of the peak metabolic rate and the magnitude of the SDA effect.

Animal Feed↗

PubMedDate not supplied

A quantum chemical and statistical study of flavonoid compounds (flavones) with anti-HIV activity.

The molecular orbital semi-empirical method AM1 was employed to calculate a set of molecular properties (variables) of 22 flavonoid compounds (flavones) with anti-HIV-1 activity and nine new compounds were proposed for anti-HIV-1 activity prediction. Pattern recognition techniques, principal component analysis (PCA), hierarchical cluster analysis (HCA), stepwise discriminant analysis (SDA) and K-nearest neighbor (KNN), were employed in order to reduce dimensionality and investigate which subset of variables could be more effective for classifying the flavones according to their degree of anti-HIV-1 activity. The PCA, HCA, SDA and KNN studies showed that the variables log P (partition coefficient), molecular volume (VOL) and electron affinity (EA) are responsible for the separation between anti-HIV-1 active and inactive compounds. The prediction study was done with a new set of nine analog compounds by using the PCA, HCA, SDA and KNN methods and only one of them was predicted as active against HIV-1.

Anti-HIV Agents↗

PubMed2005

Application of Phi29 DNA polymerase mediated whole genome amplification on single spores of arbuscular mycorrhizal (AM) fungi.

Genetic analysis of arbuscular mycorrhizal (AM) fungi relies on analysis of single spores. The low DNA content makes it difficult to perform large scale molecular analysis. We present the application of Phi29 DNA polymerase mediated strand displacement amplification (SDA) to genomic DNA extracted from single spores of Glomus and Gigaspora species to address this problem. The genome coverage of the SDA process was evaluated by PCR amplification of the beta-tubulin1 gene and part of the rDNA cluster present in AM fungi. The fidelity of SDA was evaluated further by sequencing the Glomus intraradices ITS1 variants to detect the four ITS1 variants previously identified for this fungus.

Bacillus Phages↗

PubMed2006

Vagal afferents mediate the feeding response to mercaptoacetate but not to the beta (3) adrenergic receptor agonist CL 316,243.

To evaluate the role of subdiaphragmatic vagal afferents in the anorectic response to peripheral administration of the highly selective beta(3)-adrenergic receptor agonist CL 316,243 (CL), we tested the ability of intraperitoneal (IP) injections of CL to inhibit feeding in rats with subdiaphragmatic vagal deafferentation (SDA, n=13) or sham surgeries (SHAM, n=13). Doses of 10, 100 and 1000 ng/kg CL significantly reduced feeding by statistically similar amounts in SHAM and SDA rats. One hour after IP injection, each dose of CL also significantly increased plasma concentrations of free fatty acids and beta-hydroxybutyrate, an indicator of hepatic fatty acid oxidation (FAO), whereas 6h after injection only the two highest CL doses increased plasma beta-hydroxybutyrate. In contrast, peripheral administration of the FAO inhibitor mercaptoacetate (MA, 45.6 mg/kg IP) stimulated feeding in SHAM but not in SDA rats, extending previous data suggesting a necessary role of vagal afferents in the feeding-stimulatory effect of FAO inhibition. We conclude that subdiaphragmatic vagal afferents are essential for the feeding-stimulatory action of MA but not for the anorectic action of peripheral CL and that CL-induced increase in hepatic FAO is not essential for its feeding-inhibitory effect.

3-Hydroxybutyric Acid↗

PubMed2005

Tubular and glomerular proteinuria in diagnosing chronic allograft nephropathy with relevance to the degree of urinary albumin excretion.

The diagnosis of chronic allograft nephropathy (CAN) is based on pathological examination according to Banff 97 schema. The aim of the study was to evaluate the usefulness of tubular and glomerular proteinuria for noninvasive recognition of CAN. One hundred and thirty renal allograft recipients (at least 90 days after transplantation) who had undergone diagnostic allograft biopsy were included in the study. Beta2-microglobulin, alpha1-microglobulin, albumin, immunoglobulin G, total protein, and creatinine concentrations were obtained from the second morning urine specimen. Raw data and values calculated per 1 g of creatinine excreted in urine along with time after transplantation, serum creatinine, and its change over a period of 2 months prior to biopsy were taken for analysis. Urine proteins were measured using a nephelometric method. Statistical calculations were performed using MANOVA and stepwise discriminant analysis (SDA). Statistical diagnosis and staging of CAN matched the pathological method in 68% of a preliminary SDA. Therefore patients were divided into normoalbuminuric, microalbuminuric, and macroalbuminuric groups. There was no significant differences between protein excretion, except alpha1-microglobulinuria (CAN 0 vs 2, P = .018; CAN 1 vs 2, P = .041), beta2-microglobulinuria (CAN 0 vs 2, P = .026; CAN 1 vs 2, P = .0033), and total proteinuria (CAN 0 vs 2, P = .042) in the normoalbuminuric group. Nevertheless, diagnoses obtained using SDA were 89%, 91%, and 92% identical to the results of pathological examinations, for normoalbuminuric, microalbuminuric, and macroalbuminuric groups, respectively. In conclusion, tubular and glomerular proteinuria measurements may be useful for a noninvasive CAN diagnosis and staging only with regard to degree of urinary albumin excretion.

Albuminuria↗

PubMed2001

Feeding behavior during sialodacryoadenitis viral infection in rats.

Sialodacryoadenitis (SDA) is a highly contagious common viral infection in rats, akin to mumps in humans. Anorexia occurs during such viral infection. But the pattern of the decrease in food intake (a decrease in either meal size and meal number or both) during spontaneous viral infection has not been previously characterized. We observed the onset of anorexia and an abnormal feeding pattern during an opportunistic SDA viral infection in our rat colony. We thus studied seven male rats. Before the viral infection there was a positive association between food intake and meal number (P<.05). After infection food intake decreased by 68%. This occurred via a significant decrease in meal size (by 69%) (P<.05); and a nonsignificant decrease in meal number (P=.71). This pattern of decreased food intake is similar to that occurring during indomethacin-induced ulcerative ileitis, where we previously measured an increase in plasma tumor-necrosis factor (TNF)-alpha. Anorexia in response to bacterial lipopolysaccharide administration, which is also linked to plasma TNF-alpha, is however, caused only via a decrease in meal number. The differences in the decrease in the feeding pattern between the SDA viral and a bacterial infection suggest that factors other than TNF-alpha alone play a significant role in the mechanism of anorexia during a viral infection.

Animals↗

PubMed2003

Selective effects of vagal deafferentation and celiac-superior mesenteric ganglionectomy on the reinforcing and satiating action of intestinal nutrients.

The role of vagal afferents and splanchnic fibers in nutrient-induced flavor conditioning and feeding suppression was determined. Male rats were fitted with intraduodenal (ID) catheters and given subdiaphragmatic vagal deafferentation (SDA), celiac-superior mesenteric ganglionectomy (CGX), combined (COM) treatments, or sham surgery. In separate conditioning trials, they were trained to drink (30 min/day) flavored saccharin solutions paired with concurrent ID infusions of 8% maltodextrin or water and 3.55% corn oil or water. Experiment 1 revealed that SDA and sham rats showed equal preferences for the nutrient-paired flavors over the water-paired flavors. In contrast, SDA rats, unlike sham rats, failed to suppress their intake of a palatable fluid when infused intraduodenally with maltodextrin or corn oil. Experiment 2 revealed that CGX, COM and sham rats all developed preferences for the maltodextrin-paired flavor, although CGX alone or COM attenuated the conditioned preference. CGX and COM treatments also attenuated or blocked the feeding inhibitory actions of ID nutrient infusions. These findings along with prior data indicate that gut vagal afferents and splanchnic nerves are not essential for flavor-nutrient preference conditioning, whereas both vagal afferents and splanchnic nerves are implicated in carbohydrate- and fat-induced satiation.

Afferent Pathways↗