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PubMed1998

Stromelysin 3 is overexpressed in human pancreatic carcinoma and regulated by retinoic acid in pancreatic carcinoma cell lines.

BACKGROUND: Matrix metalloproteinases play an important role in the control of local tumour growth and metastasis of human pancreatic cancer. AIMS: To examine expression of recently discovered stromelysin 3 (STR-3) in human pancreatic cancer and pancreatic carcinoma cell lines and to investigate their regulation by retinoids. METHODS: STR-3 expression was examined by immunohistochemistry in 21 human pancreatic carcinomas. Expression of STR-3 and regulation by retinoids was assessed in five human pancreatic carcinoma cell lines using western and northern blotting as well as nuclear run on assays. RESULTS: There was pronounced overexpression of STR-3 in 17 of 21 (80.9%) pancreatic carcinoma specimens. STR-3 expression was predominantly located in peritumourous stromal cells. Six of 21 (28.5%) carcinomas also revealed STR-3 expression in epithelial tumour cells whereas no STR-3 expression was observed in non-transformed pancreas. All five pancreatic carcinoma cell lines expressed STR-3 mRNA and protein. Furthermore, retinoid treatment results in a time and dose dependent inhibition of STR-3 protein expression. This inhibition seems to be post-transcriptional as neither STR-3 gene transcription nor mRNA steady state concentrations were affected by retinoids. CONCLUSIONS: STR-3 overexpression in stromal as well as epithelial elements during pancreatic carcinogenesis might contribute to the aggressive local growth and metastasis of pancreatic cancer and can be therapeutically targeted by retinoids.

Blotting, Western↗
PubMed1986

Scotopic threshold response of proximal retina in cat.

We describe a new response in proximal retina of cat that is present under scotopic conditions, clearly differs from PII (b-wave and DC component) and contributes a negative potential at threshold to the diffuse electroretinogram (ERG). We have termed this response the scotopic threshold response (STR). Extracellular potentials evoked in response to circular spots of light at dark-adapted threshold, and with dim backgrounds, were recorded with microelectrodes placed intraretinally at different depths. The dark-adapted response of proximal retina (STR) consisted of a graded negative potential to the onset of illumination that maintained amplitude during illumination and decayed back toward the base line at stimulus offset without evidence of a negative-going off response. It thereby differed in form from the photopic M-wave response of proximal retina, which has a negative-going off response. It also did not exhibit spatial tuning, simply increasing in size with stimulus area. In addition, the STR appears to be a rod-driven response whose threshold approximates that of the most sensitive ganglion cells in cat, whereas the M-wave is a much higher threshold cone-driven response. The STR could be clearly distinguished from PII on the basis of its form, depth-distribution, and dynamic range. For example, the STR had its maximum amplitude in the proximal retina at 17% retinal depth, whereas scotopic PII had its maximum in the distal retina at 48% retinal depth. Also, the STR had a lower threshold than PII intraretinally and saturated well below the level of saturation of scotopic PII (rod saturation). By analogy to the PNR and M-wave, the STR is hypothesized to represent either an extracellular voltage arising from proximal retinal neurons or Müller cell responses to K+ released by these neurons. Recording in the vitreous, near the retinal surface, showed that the STR always had a negative polarity. The polarity reversal of the STR at 50-60% retinal depth (from negative, proximal to positive, distal) suggested the presence of a sink proximal to the reversal point and a source distal to it. We also recorded the vitreal ERG in response to diffuse illumination of the dark-adapted retina. The STR could be clearly identified in the scotopic ERG as a threshold negative potential that had been observed previously in the mammalian ERG. The STR differs, therefore, from PII (b-wave and DC component) that is a higher threshold positive component in the diffuse ERG.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2001

Number, origins, and chemical types of rat pallidostriatal projection neurons.

The dorsal globus pallidus (GP) receives major inputs from the dorsal neostriatum (Str), the subthalamic nucleus (STN) and the dorsal thalamus. The GP projects to multiple basal ganglia nuclei. One of the GP projection sites is the Str. The pallidostriatal projection has been considered minor. However, several recent studies have suggested that this projection is heavier than previously thought and that it might play a significant role in controlling the activity of the Str. To reveal more details of this projection, we examined the number of GP neurons that participated in the projection, their origins in the GP and their immunoreactivity for the calcium binding protein parvalbumin (PV), by using a combination of Fluoro-Gold (FG) retrograde labeling and immunohistochemical methods. Immunostaining for the calcium binding protein calbindin-28K (CaBP) was used to identify the CaBP-poor sensorimotor and CaBP-rich associative Str regions and the corresponding CaBP-poor middle, CaBP-rich border, and the caudomedial GP regions. The CaBP-poor dorsolateral Str region occupies a small portion of the Str, whereas the CaBP-poor middle GP region occupies a large portion of the GP. The immunostaining for neuron-specific nuclear protein (NeuN) was used to visualize neurons that were immunonegative for FG or PV. Cell counts revealed that the middle GP region contained a higher density of neurons and also a higher percentage of PV-positive neurons than the border and caudomedial regions of the GP. These observations suggested that the GP is involved more in sensorimotor function than associative function. Approximately 40% of neurons in the CaBP-poor middle GP region project to the CaBP-poor part of the dorsolateral Str. Approximately 30% of the neurons in both the CaBP-rich border and the caudomedial GP regions project to the CaBP-rich Str region. More than 40% of the pallidostriatal neurons in CaBP-poor middle GP region are PV-positive, whereas most of those in CaBP-rich GP regions are PV-negative. It was estimated from the cell count data that most of the PV-negative neurons in all three regions of the GP project to the Str. The results indicate that the sensorimotor and associative territories of the Str have reciprocal projections between corresponding territories of the GP. The involvement of a large number of GP neurons suggested that the pallidostriatal projection should be taken into account in the analysis of functional roles of the basal ganglia.

Animals↗
PubMed2004

[Prediction of outcome in ST elevation myocardial infarction by the extent of ST segment deviation recovery. Which method is best?].

UNLABELLED: Simple and rapid measures are needed for timely assessment of the quality of reperfusion therapy early after fibrinolysis in acute STEMI. Sum ST segment elevation resolution (sum STR) categorized into the three groups of low risk (complete ST resolution), medium risk (partial ST resolution), and high risk (no ST resolution) has become an established method to predict infarct size, left ventricular function, epicardial vessel patency, and mortality. However, measurement of the sum of ST elevation from all leads of repeated ECG's is time-consuming. For routine practice more simple measures are needed. This report summarizes recent findings on direct comparisons between different modes of evaluation of ST segment deviation recovery employed for risk stratification in large-scale mortality trials. With respect to predictive accuracy combined with simplicity, two methods were superior to the conventional model of sum STR: 1) ST segment deviation resolution in only the one ECG lead showing the maximal deviation (single lead STR), and 2) the existing ST segment deviation in the single ECG lead of maximum deviation present 90 or 180 min after start of fibrinolysis (max STE). In multivariate analyses the ST segment deviation recovery models including sum STR were significant independent predictors of short- and long-term mortality. In receiver-operating characteristic (ROC) curves for predicting mortality the analysis of single lead STR and max STE performed better than sum STR. After categorization into risk groups patients are best classified by max STE. With an ECG recorded at 90 min in 2719 patients, the proportion of patients of sum STR, single lead STR, and max STE were 40, 34, and 43% in the low risk groups, and 24, 31, and 25% in the high risk groups. Cardiac mortality rates at 30 days were 2.0, 1.2, and 1.0% in low risk versus 9.6, 10.3, and 12.8% in the high risk groups, respectively. Long-term mortality with a followup of 5 years was best predicted by max STE risk groups. CONCLUSION: Single lead STR and max STE are very simple, inexpensive, non-invasive, and highly reliable measures which provide very strong early prognostic information. The relationship between degree of ST segment deviation recovery and subsequent mortality is remarkably consistent. Both methods perform better than sum STR in predicting mortality. They can be used for very early risk stratification and can form a basis for an individual treatment of patients after fibrinolysis for STEMI within 6 hours of symptom onset. Of the two methods max STE is even simpler to use and has better accuracy in predicting outcome.

Aged↗
PubMed1990

The effects of strychnine on the regulation of voltage-dependent calcium channels by dihydropyridines in brain and heart.

The effects of strychnine (STR) were investigated on K(+)-stimulated 45Ca2(+)-uptake into mouse brain neurons, the contractile activity of spontaneously beating rat atria and on [3H]nitrendipine and [3H]BAY K 8644 binding to dihydropyridine calcium channel antagonist and agonist binding sites on brain and cardiac membranes. STR (10(-6)-10(-4) M) had no effect on neuronal 45Ca2(+)-uptake. When combined at equimolar concentrations (10(-5) M), STR and nifedipine produced a potent (nM) inhibition (40%) of neuronal 45Ca2(+)-uptake. In the spontaneously beating rat atria, STR produced a dose-dependent (10(-7)-3 x 10(-4) M) decrease in chronotropy but did not affect inotropy. STR (10(-4) M) completely inhibited the positive chronotropic, but did not affect the positive inotropic effects of (-)-S-BAY K 8644. [3H]Nitrendipine and [3H]BAY K 8644 binding to brain and cardiac membranes was enhanced by STR in a concentration-dependent manner (EC50 8 X 10(-6) M). Scatchard analysis revealed that STR increased the affinity (decreased the Kd) of [3H]BAY K 8644 to a greater degree than that of [3H]nitrendipine for dihydropyridine binding sites. STR decreased the Kd of [3H]nitrendipine binding by increasing and decreasing the microassociation and microdissociation constants respectively. STR enhanced [3H]nitrendipine binding to the same extent in the cerebral cortex, striatum, hippocampus, cerebellum, brainstem and spinal cord. The enhancement of [3H]nitrendipine binding in brain was completely inhibited by Ca2+ and partially inhibited by Na+ in a concentration-dependent manner. Glycine (10(-2) M) did not affect the STR enhancement of [3H]nitrendipine binding.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed2001

Stimulation of either cholecystokinin receptor subtype reduces while antagonists potentiate or sensitize a morphine-induced excitatory response.

Cholecystokinin peptides (CCK) have been shown to antagonize many opioid-mediated effects. The present study was undertaken to determine whether peripheral injections of cholecystokinin sulphated octapeptide (CCK8), cholecystokinin tetrapeptide (CCK4), the CCK(1) (lorglumide) and the CCK(2) (PD-135,158 and LY-225910) receptor antagonists can influence a classic morphine excitatory effect, i.e. the display of Straub tail reaction in mice (STR). A total of 570 female Balb/C mice were tested. Experiment 1 was undertaken to determine whether i.p. injections of CCK8 or CCK4 can influence STR. Each animal was treated with i.p. injections of saline or CCK8 (10 and 20 nmol/kg) or CCK4 (20 and 40 nmol/kg). After 30 min all animals received an i.p. injection of morphine hydrochloride (10.0 mg/kg). The highest doses of both CCK8 (35% STR) and CCK4 (40% STR) significantly reduced STR as compared to saline (85% STR) treated mice (Fisher test; P < 0.01). In experiment 2 each animal was treated with ip injections of saline or 1.0 mg/kg lorglumide or PD-135,158 fifteen minutes before an injection of morphine at doses ranging from 1.0 to 50.0 mg/kg. In experiment 3 animals were treated with injections of saline, 0.1 or 10.0 mg/kg lorglumide or LY-225910 before an injection of a fixed MC dose (2.0 mg/kg). Both lorglumide and PD-135,158 induced a significant shift to the left in the morphine dose-response curves as well as a significant decrease in ED50 of the STR. ED50 for lorglumide was significantly lower than ED50 for PD-135,158. Both doses of lorglumide and the highest dose of LY-225910 significantly increased the percent of animals displaying STR. Experiment 4 was undertaken to determine whether repeated peripheral injections of morphine or the morphine-potentiating agents CCK(1) (lorglumide) and the CCK(2) (LY-225910) receptor antagonists can induce morphine sensitization. Each animal was treated with 5 daily i.p. injections of saline (control group), 1.5 mg/Kg morphine hydrochloride (group morphine), and 1.0 mg/Kg lorglumide (group LOR) or LY-225910 (group LY). One, two, three and four weeks after the last treatment day, all animals were challenged with one i.p. injection of morphine (1.5 mg/Kg). The morphine, LOR groups and group LY showed a significant increase in percentage of animals displaying STR. These data demonstrate that the blockade of endogenous CCK actions leads to morphine sensitization probably through both CCK receptors. The present data are consistent with the antagonistic effects of CCK and opioids in the control of morphine-induced STR. In addition, these results suggest that both CCK receptors are involved in the modulatory effects of CCK on this morphine effect.

Analgesics, Opioid↗
PubMed1998

Milacemide, a glycine pro-drug, inhibits strychnine-allodynia without affecting normal nociception in the rat.

The blockade of spinal glycine receptors with intrathecal (i.t.) strychnine (STR) produces reversible, segmentally localized allodynia in the rat. The purpose of this study was: (1) to investigate the effect of the anticonvulsant agent, milacemide, a glycine pro-drug on STR-allodynia; (2) to compare this effect with that of milacemide on normal nociception (without STR); and (3) to determine the sensitivity of the anti-allodynic effect of milacemide to pretreatment with selective monoamine oxidase (MAO)-A (clorgyline) and MAO-B (L-deprenyl) inhibitors. Male Sprague-Dawley rats, fitted with chronic i.t. catheters, were lightly anesthetized with urethane. Hair deflection (HD) evoked maximum changes in blood pressure and heart rate were recorded from left carotid artery, and cortical electroencephalographic (EEG) activity was continuously monitored using subdermal needle electrodes before and after i.t. STR (40 microg). Rats were pretreated with a single intravenous (i.v.) injection of milacemide (100-600 mg/kg), 1 h before i.t. STR. To sustain the allodynic state, STR was injected every hour for up to 4 h. HD was applied to the affected dermatomes (2 min duration) using a cotton-tipped applicator at 5-min intervals for the duration of the STR effect. Normally innocuous HD elicited a marked increase in mean arterial blood pressure and heart rate, an immediate motor responses, and desynchronisation of EEG when applied to the cutaneous dermatomes affected by i.t. STR. Milacemide (100-600 mg/kg, i.v.) dose-dependently inhibited the heart rate and pressor responses (ED50 = 398 mg/kg; 95%CI = 196-873) and the motor responses (ED50 = 404 mg/kg; 95%CI = 275-727). Maximum inhibition was observed approximately 2 h after i.v. injection. The duration of action ranged from 3 h (400 mg/kg) to 4 h (600 mg/kg). Milacemide had no effect on the percent synchrony in the EEG. At the time of maximum inhibition of STR-allodynia (2 h post-infusion), responses evoked by noxious pinch were unaffected by milacemide. Pretreatment with L-deprenyl (3 mg/kg, i.p.), but not clorgyline (10 mg/kg, i.p.) significantly blocked the anti-allodynic effect of milacemide (600 mg/kg i.v). These data indicate that i.v. milacemide significantly attenuates the allodynia arising from spinal glycine receptor blockade, and are consistent with: (1) the selective modulation of low threshold afferent input by STR-sensitive, glycine interneurons in the rat spinal cord; and (2) the pharmacological actions of milacemide as a glycine pro-drug.

Acetamides↗
PubMed1997

The screening of 13 short tandem repeat loci in the Chinese population.

Population studies of 13 short tandem repeat (STR) loci were carried out on Chinese in Taiwan. The STR loci included HUMF13B, HUMF13A01, HUMFES/FPS, HUMFABP, HUMPLA2A1, HUMTPOX, HUMTH01, HUMVWFA31/A, HUMCSFIPO, HUMLPL, HUMGPP3A09, HUMCYAR04 and HUMCD4. DNA samples from 100 unrelated individuals were screened. The STR allele patterns were detected by the fluorescence detector of an automated DNA sequencer. Two PCR amplifications were performed for each STR locus in this study. The first PCR amplification strategy used 26 base pairs of the T7 sequence extension in the 5' end of the forward primer of each STR locus. The second PCR amplification used a dye-labeled T7 primer instead of the forward primer in the first PCR amplification, and the first PCR products as template to produce fluorescent dye-labeled PCR products. PCR products of different STR loci with overlapping allele sizes could be detected in the same lane of the polyacrylamide gel on an automated DNA sequencer using different colored dye-labeled T7 primers. There was no need to directly conjugate the fluorescent dye to individual STR primers. The PCR products were obtained using 2 ng of template DNA in 25 microliters of PCR reaction mixture. No deviations from the Hardy-Weinberg equilibrium were observed for the 13 STR loci. The distributions of these STR alleles were different from those of Caucasians or Blacks. The probability of matching from the combination of the 13 STR loci was 5.9 x 10(-10) for our Chinese population. However, HUMF13B, HUMLPL and HUMCD4 loci were not as highly polymorphic as observed in other populations.

Alleles↗
PubMed2005

Evaluation of tRNA intergenic spacer length polymorphism analysis as a molecular method for species identification of streptococcal isolates from bovine mastitis.

Sixty-nine bovine mastitis streptococci belonging to the species Str. agalactiae (n = 13), Str. dysgalactiae (n = 16), Str. canis (n = 22), Str. uberis (n = 20) and Str. parauberis (n = 4) and six reference strains of the five streptococcal species were examined for their tRNA gene intergenic length polymorphism (tDNA-ILP) fingerprint pattern. Epidemiologically unrelated isolates from bovine mastitis cases were selected by macrorestriction analysis with pulsed-field gel electrophoresis (PFGE). Their results were compared with those obtained from biochemical and serological studies and with those obtained by PCR-mediated identification amplifying species-specific gene segments of the five streptococcal species. According to the present results tDNA-ILP allowed a correct identification of all Str. agalactiae, Str. uberis and Str. parauberis strains investigated also including the reference strains of each species showing species-specific banding pattern. However, all Str. dysgalactiae ssp. dysgalactiae and all Str. canis strains appeared with an undistinguishable pattern which did not allow an identification of the species.

Animals↗
PubMed2000

Monitoring of residual disease and guided donor leucocyte infusion after allogeneic bone marrow transplantation by chimaerism analysis with short tandem repeats.

In this study, we analysed the chimaeric status of peripheral blood leucocytes (PBLs) in recipients of allogeneic bone marrow transplantation (BMT) with the use of short tandem repeat (STR) microsatellite markers for monitoring the efficacy of BMT and donor leucocyte infusions (DLIs). A set of four STR markers was used with a highly discrimative capacity between individuals. STRs were detected by polymerase chain reaction (PCR) and were analysed by gene scanning (STR-GS). Between June 1990 and December 1998, 52 patients treated with BMT for chronic myeloid leukaemia (CML) were analysed. Seventeen patients relapsed after BMT and two patients never achieved remission after BMT. Fourteen of the 17 patients achieved a complete donor chimaerism after BMT, as detected by the presence of only donor STR-GS fragments, and in three cases a weak recipient STR-GS signal remained persistently detectable after BMT. A reappearance or increase of recipient STR-GS signals was indicative of relapse, which was mostly detected by STR-GS several months before relapse was diagnosed clinically. Nineteen patients were treated with DLI for reappearance of CML after BMT which resulted in complete remission in 17 patients, concordant with the disappearance of recipient STR-GS signals. More importantly, DLI treatment could be guided based upon the STR-GS data, which prevented unnecessary extra DLI courses that could cause toxicity. This study indicates that STR-GS is an effective and reliable method for monitoring BMT recipients.

Adult↗
PubMed2006

The HIV-1 clade C promoter is particularly well adapted to replication in the gut in primary infection.

OBJECTIVE: Coinfection of rhesus macaques with human/simian immunodeficiency virus chimeras harbouring the minimal core-promoter/enhancer elements from HIV-1 clade B, C and E viral prototypes (STR-B, STR-C and STR-E) revealed a remarkable dichotomy in terms of spatio-temporal viral replication. The clade C chimera (STR-C) predominated in primary infection. The present study was aimed at identifying the origin of STR-C plasma viraemia at this infection phase. DESIGN: By competing isogenic viruses differing only in their promoters, it was possible to identify subtle phenotypical differences in viral replication kinetics and compartmentalization in vivo. METHODS: Two rhesus macaques were coinfected by the three STR chimeras and the relative colonization of different compartments, particularly blood and stool, was determined for each chimera. Moreover, growth competition experiments in thymic histocultures enriched in interleukin (IL)-7 were performed and relative percentages of chimeras were estimated in supernatants and thymocytes lysates at different time points. RESULTS: It is demonstrated here that at the peak of primary infection, preferential replication of STR-C was supported by the gut-associated lymphoid tissue (GALT), an IL-7 rich microenvironment. This was shown by the correlation of the RNA viral genotype in blood and stools, compartments directly draining virions from the GALT. Thymic histocultures confirmed that replication of STR-C is particularly susceptible to this cytokine, compared to its STR-B and STR-E counterparts. CONCLUSIONS: These data show that the GALT cytokine network may well favour HIV-1 clade C replication during primary infection. This could result in enhanced transmission.

Animals↗
PubMed1993

Therapy of experimental murine brucellosis with streptomycin alone and in combination with ciprofloxacin, doxycycline, and rifampin.

The in vivo efficacy of streptomycin (STR), doxycycline (DOX), rifampin (RIF), ciprofloxacin (CIP), and their combinations was evaluated for a Brucella melitensis experimental infection in a mouse model. Animals were infected with 2 x 10(4) to 4 x 10(4) CFU of B. melitensis intraperitoneally on day 0 and were randomized to receive, starting on day 7, STR alone at 75, 150, or 300 mg/kg of body weight per day intraperitoneally or DOX at 6 mg/kg/day orally, RIF at 3 mg/kg/day orally, or CIP at 200 mg/kg/day orally, each of the last three drugs alone or in combination with STR at 75, 150, or 300 mg/kg/day, for 14 days. Therapy failure (defined as nonsterile spleens) was observed in all animals treated with STR at all doses and with CIP given as monotherapy. Mean log CFU isolated from the spleens remaining infected following monotherapy with STR or CIP were not different from those in control mice. RIF at a low dose did not have an effect on cure rates; however, a reduction in CFU relative to the CFU in untreated animals was obtained. DOX at low levels achieved a 35% cure rate and a reduction in CFU in animals not cured. All animals treated with DOX or RIF combined with any STR dose were cured, but none of the animals receiving the STR-CIP combinations was cured, and the splenic CFU remained similar to those in the controls. These results demonstrate that the combinations DOX-STR and RIF-STR are synergistic against B. melitensis, while the combination STR-CIP is indifferent and ineffective in the management of acute murine brucellosis. The results also appear to support the clinical superiority of combination drug therapy over monotherapy.

Animals↗
PubMed1997

Coordinate expression of matrix metalloproteinase family members in the uterus of normal, matrilysin-deficient, and stromelysin-1-deficient mice.

The expression patterns of matrix metalloproteinase (MMP) family members during the murine estrous cycle and postpartum uterine involution were analyzed, and the consequence of removing specific MMPs during uterine functions was determined using mice deficient in either matrilysin (MAT) or stromelysin-1 (STR-1). In wild-type animals, MAT, STR-1, STR-2, STR-3, and gelatinase A were consistently expressed during the most active phases of the estrous cycle, estrus and proestrus. The messenger RNA for these MMPs as well as collagenase-3 and the tissue inhibitors of metalloproteinases were also expressed during uterine involution, as determined by Northern analysis and in situ hybridization. Notably, MAT, STR-2, and collagenase-3 messenger RNA levels were elevated at early times of involution and rapidly decreased with time, whereas the transcripts for other MMPs remained elevated throughout the involution process. Involution proceeded normally in mice lacking MAT or STR-1; however, the expression of STR-1 and STR-2 was dramatically up-regulated in MAT nullizygous mice, and the expression of MAT and STR-2 was moderately up-regulated in STR-1-deficient animals. We conclude that the concerted action of several MMPs is likely to play an important role in the remodeling of the postpartum uterus, and that mechanisms that compensate for the loss of a specific MMP during this process appear to exist.

Animals↗
PubMed2005

Comparison of variable number tandem repeat and short tandem repeat genetic markers for qualitative and quantitative chimerism analysis post allogeneic stem cell transplantation.

BACKGROUND: Analysis of donor chimerism has become a routine procedure for the documentation of engraftment after allogeneic hematopoietic stem cell transplantation. Quantitative analysis of chimerism kinetics has been shown to predict graft failure or relapse. In this study, we compared the use of variable number tandem repeats (VNTR) and short tandem repeats (STR) as polymorphic genetic markers in chimerism analysis. This study included qualitative and quantitative assessment of both techniques to assess informative yield and sensitivity. PATIENTS AND METHODS: We analyzed 206 samples representing 40 transplant recipients and their HLA-identical sibling donors. A panel of six VNTR loci, 15 STR loci and 1 sex chromosome locus was used. Amplified VNTR products were visualized in an ethidium bromide-stained gel. STR loci were amplified using fluorescent primers, and the products were analyzed by capillary electrophoresis. RESULTS: VNTR and STR analysis gave comparable qualitative results in the majority of cases. The incidence of mixed chimerism (MC) by STR analysis was 45% compared to 32% in cases evaluated by VNTR analysis. STR markers were more informative; several informative loci could be identified in all patients. Unique alleles for both patient and donor could be identified in all patients by STR versus 32/40 by VNTR analysis. The STR markers were also more sensitive in the detection of chimerism. The size of VNTR alleles and differences between the size of donor and recipient VNTR alleles affected the sensitivity of detection. With both techniques, quantitative assessment of chimerism showed some discrepancies between the estimated and the calculated percentage of donor DNA. Discordance between the two estimates was observed in 8/19 patients with MC. However, sequential monitoring of the relative band intensity of VNTR alleles offered some insight into the direction of change in engraftment over time. CONCLUSION: The higher yield of informative loci with STR and the automated measurement of amplified STR 103 products offered the advantages of more rapid and accurate quantitative assessment of chimerism. The choice between these two techniques depends on the need for quantitative or qualitative information, the availability of equipment, and the cost.

Chimerism↗
PubMed2006

[Effects study of standardized tertiary rehabilitation on promoting of the neurological functions in stroke patients with hemiplegia].

OBJECTIVE: To investigate the effects of standardized tertiary rehabilitation (STR) on promotion of the neurological functions in stroke patients with hemiplegia. METHODS: A large-sample, multi-center, randomized, controlled prospective study was conducted nationwide. 1209 stroke patients with hemiplegia, aged 40 - 80, within 11 d +/- 5 d after the onset, stabilized in vital life signs in the past one week, of 20 hospitals affiliated to medical colleges or of hospitals at the provincial level in mainland China were randomly allocated to one of the 2 baseline data-matched groups: control group (n = 599, 446 with cerebral infarction and 153 with cerebral hemorrhage) receiving routine intervention of internal medicine, and STR group (n = 610, 455 with cerebral infarction and 155 with cerebral hemorrhage), receiving routine intervention and STR including physical therapy and occupational therapy (OT) in addition. Evaluation was conducted by the time of enrollment, and by the ends of the first, third, and sixth months by the national clinical neurological function defects (CNFD) scoring. RESULTS: There were no significant differences in the baseline indexes between these 2 groups. The scores of CNFD at enrollment of the patients with cerebral infarction in the STR group was 21.55, not significantly different from that of the patients with cerebral infarction in the control group (22.16), however, the scores of these patients with cerebral infarction in the STR group by the ends of the first, third, and sixth months were 16.32, and 11.48, and 8.63 respectively, all significantly lower than those of the corresponding patients in the control group (18.95, 15.57, and 13.78 respectively, all P < 0.01). The CNFD score at enrollment of the patients with cerebral hemorrhage in the STR group was 23.27, not significantly different from that of the corresponding patients in the control group (24.36), however, the scores of the patients with cerebral hemorrhage in the STR group by the ends of the first, third, and sixth months were 17.61, 11.20, and 7.95 respectively, all significantly lower than those of the corresponding patients in the control group (20.36, 15.01, and 13.57 respectively, all P < 0.01). By the end of the sixth month, the CNFD score of the cerebral infarction and hemorrhage patients in the STR group were improved by 12.79 and 15.23, both higher than the corresponding patients in the control group (8.30 and 10.65 respectively). The trend of decrease of CNFD score moved faster in the STR group than in the control group, especially at the early stage. CONCLUSION: STR significantly helps improved the neurological function in the patients with cerebral stroke accompanied with hemiplegia.

Adult↗
PubMed1993

Effect of prefrontal cooling on levels of glutamate, DOPAC and 5-HIAA in nucleus accumben and striatum in vivo.

The effect of pre-frontal cooling of the cortex (PFC) on monoaminergic metabolites (DOPAC and 5-HIAA) and excitatory amino acids (glutamate (Glu) and aspartate (Asp)) was investigated using microdialysis in rat striatum (Str) and nucleus accumben (NuAc). The cooling profoundly decreased Glu and elevated DOPAC and 5-HIAA in both Str and NuAc for a long period of time. However, the time course of the detection of each compound appeared different between Str and NuAc. In NuAc, Glu decreased rapidly to reach the plateau in about 30 min; however, in Str, Glu decreased steadily and reached the plateau in about 60 min. The time courses of elevation of 5-HIAA and DOPAC were also quite different between Str and NuAc; there was more profound change observed in Str than in NuAc. The level of DOPAC exhibited a similar period of increase (7.14 min) in NuAc and Str but in NuAc it decreased toward the baseline much earlier (about 2h) than Str (over 3 h). The level of 5-HIAA in Str showed a period of rapid increase to a plateau (less than 42 min) but a trend of decrease started before the termination of sampling. In NuAc, in contrast, 5-HIAA increased slowly (more than 48 min) and maintained a plateau until termination of sampling. We conclude that PFC cooling may have caused an increase in Glu secretion and resulted in the release of the tonic restraint on DA terminals in both Str and NuAc, thus increases the synaptic turnover of monoamines.(ABSTRACT TRUNCATED AT 250 WORDS)

3,4-Dihydroxyphenylacetic Acid↗
PubMed1996

Scotopic threshold response of the electroretinogram of dogs.

OBJECTIVE: To study characteristics of the scotopic threshold response (STR) in dogs. DESIGN: We recorded the electroretinogram (ERG) in anesthetized Beagles under several recording conditions to examine characteristics of the STR. ANIMALS: Seven clinically normal Beagles. PROCEDURE: Beagles were paralyzed by continuous i.v. administration of muscle relaxant and were artificially ventilated with a gas mixture of nitrous oxide and oxygen during experiments. Corneal ERG were recorded, using full-field white stimuli, under several recording conditions. RESULTS: The canine STR was a simple corneal negative response elicited by dim stimuli below the b-wave threshold. Maximal amplitude of the canine STR was approximately 50 to 120 microV, much larger than those of other animal species, such as human beings, monkeys, and cats. Peak latency of the maximal STR was approximately 75 to 96 milliseconds. The spectral sensitivity of the canine STR corresponded to that of the canine rod visual pigment. The canine STR was completely eliminated by dim background light, which had no suppressive effect on the a- and b-waves. By using such dim background light, we could record the photoreceptor-derived a-wave in the ERG intensity series. CONCLUSIONS: In Beagles, the STR strongly influences the ERG waveforms recorded under the dark-adapted condition. Special attention may be necessary in using the dark-adapted ERG to assess the retinal photoreceptor function. Canine STR is expected to be a good indicator of assessment of the proximal retinal function in dogs, and Beagles might be a good experimental animal for the study of the STR.

Acclimatization↗
PubMed1999

Efficiency of two different nine-loci short tandem repeat systems for DNA typing purposes.

BACKGROUND: Genotyping based on short tandem repeat (STR) regions is widely used in human identification and parentage testing, in gene mapping studies, and as an approach to studies on the etiopathogenesis and diagnosis of hereditary diseases. We wished to study a new analytical approach that uses capillary electrophoresis and multicolor fluorescence in place of slab gel electrophoresis. METHODS: We evaluated the efficiency for parentage and forensic purposes of the AmpFLSTR Profiler PlusTM typing kit that is used with the ABI Prism 310 Genetic Analyzer (System-2 STR), and that of a widely used panel of nine STRs analyzed with conventional slab-gel electrophoresis followed by radioactive detection (System-1 STR). System-2 STR, based on automated capillary electrophoresis and automated sizing of the alleles by Genotyper 2.0 software, was used to determine the allele frequency of the nine loci in 157 Caucasian subjects from southern Italy. On the basis of the data obtained, we submitted 40 trios to parentage testing. RESULTS: A higher median probability of paternity attribution and power of exclusion were obtained with System-2 STR vs System-1 STR: respectively, 99.99% and 99.95% (P <0.05) for attribution; and five and four excluding loci (P <0.05) for exclusion. The most informative and highly discriminating loci were D18S51, D21S11, and FGA. The combined probability of matching-by-chance for all nine STRs was 1.36 x 10(-12) for System-2 compared with 1.11 x 10(-7) obtained with the other system. The internal standard and allelic ladder of the System-2 STR facilitated accurate and precise genotyping; furthermore, System-2 STR and was faster than the conventional System-1 STR. CONCLUSIONS: The System-2 STR allows rapid testing with higher probabilities of attribution and a higher power of exclusion than with the comparison method with slab-gel electrophoresis.

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