Hemodynamic reference for neonates of different age and weight: a pilot study with electrical cardiometry.

OBJECTIVE: Electrical cardiometry (EC) is an impedance-based monitor that provides noninvasive, real-time hemodynamic assessment. However, the reference values for neonates have not been established. STUDY DESIGN: EC (Aesculon) was applied to hemodynamically stable preterm and term infants. Hemodynamic variables included cardiac output (CO), cardiac index (CI), stroke volume (SV) and heart rate (HR). Their gestational age (GA), weight and body surface area (BSA) were recorded. RESULTS: A total of 280 neonates were studied. Their GA ranged from 26(5/7) to 41(4/7) weeks, weight 800 to 4420 g and BSA 0.07 to 0.26 m(2). CO was positively correlated to GA, weight and BSA (r=0.681, 0.822, 0.830, respectively; all P<0.001). Using regression analysis, CO was most significantly correlated to BSA. Mean CI was 2.55±0.37 l min(-1) per m(2). CONCLUSION: Hemodynamic reference by EC is notably distinct among neonates of diverse maturity. CO is most closely correlated to BSA.

Increased stroke risk in Bell's palsy patients without steroid treatment.

BACKGROUND AND PURPOSE: To investigate the risk of stroke development following a diagnosis of Bell's palsy in a nationwide follow-up study. METHODS AND MATERIALS: Information on Bell's palsy and other factors relevant for stroke was obtained for 433218 eligible subjects without previous stroke who had ambulatory visit in 2004. Of those, 897 patients with Bell's palsy were identified. Over a median 2.9 years of follow-up, 4581 incident strokes were identified. We estimated hazard ratios (HR) and 95% confidence intervals [CI] with Cox proportional hazard models adjusting for age, sex, co-morbidities, and important risk factors. Standardized incidence ratio of stroke amongst patients with Bell's palsy was analyzed. RESULTS: Compared with non-Bell's palsy patients, patients with Bell's palsy had a 2.02-times (95% CI, 1.42-2.86) higher risk of stroke. The adjusted HR of developing stroke for patients with Bell's palsy treated with and without systemic steroid were 1.67 (95% CI, 0.69-4) and 2.10 (95%, 1.40-3.07), respectively. CONCLUSIONS: Patients with Bell's palsy carry a higher risk of stroke than the general population. Our data suggest that these patients might benefit from a more intensive stroke prevention therapy and regular follow-up after initial diagnosis.

Effect of oral phytoestrogen on androgenicity and insulin sensitivity in postmenopausal women.

AIM: The aim of this study was to determine and compare the effect of treatment with transdermal oestrogen and phytoestrogen on insulin sensitivity and sex hormone-binding globulin (SHBG) levels in healthy postmenopausal women. METHODS: Forty-three healthy postmenopausal women aged 68 ± 7 (mean ± SD) years who were not receiving hormonal replacement therapy completed a 3 month randomized drug therapy study. The participants were randomized to one of four groups: 0.05 mg or 0.1 mg transdermal oestrogen/day, or 40 or 80 mg oral phytoestrogen (Promensil)/day insulin sensitivity was indirectly measured using the quantitative insulin sensitivity check index (QUICKI). SHBG, total testosterone, oestradiol, and fasting glucose and insulin levels for calculation of insulin sensitivity were obtained at baseline and at monthly intervals during the 3 months of therapy. RESULTS: In healthy nondiabetic postmenopausal women, the rate of change in QUICKI was significantly different between the red clover based phytoestrogen and transdermal oestrogen groups, so that after three months of therapy, QUICKI with red clover based phytoestrogen therapy was lower than that in the transdermal oestrogen group, p = 0.01. Red clover based phytoestrogen therapy was not associated with any changes in SHBG levels whereas transdermal estrogen therapy significantly increased SHBG levels, p = 0.05. CONCLUSIONS: In contrast to transdermal oestrogen therapy, oral phytoestrogen therapy does not decrease androgenicity and is associated with a decrease in insulin sensitivity. These effects are similar to those of raloxifene and consistent with phytoestrogen's selective oestrogen receptor modulator properties.

Plasma vitamin C and tocotrienols changes in response to dietary supplementation among young male adults.

BACKGROUND AND AIMS: Intake of the antioxidant vitamins C and E lowers the oxidative stress. The study aimed to determine plasma concentrations of vitamin C and tocotrienols after supplementation of both vitamins in young male adults. MATERIALS AND METHODS: A total of 64 police recruits were randomly assigned to one of these groups: (a) 500 mg vitamin C (Vitamin C), (b) 200 mg Tocovid (Tocotrienol), (c) combination of 500 mg vitamin C and 200 mg Tocovid (Combination) or (d) placebo (Placebo) for eight-weeks of supplementation followed by six-week washout period. RESULTS: In Combination group, mean plasma vitamin C concentration significantly increased from baseline 2.86 +/- 1.19 mg/L to 10.37 +/- 1.29 mg/L and 15.63 +/- 1.27 mg/L after four- and eight-week supplementation, respectively. The corresponding figures for alpha-, delta- and gamma-tocotrienols were 9.9 +/- 2.5 ng/ml to 104.1 +/- 19.8 ng/ml and 112.8 +/- 38.0 ng/ml; 2.5 +/- 0.9 ng/ml to 29.9 +/- 7.0 ng/ml and 17.9 +/- 4.7 ng/ml; 19.2 +/- 3.1 ng/ml to 75.2 +/- 24.1 ng/ml and 161.7 +/- 49.9 ng/ml, respectively. In Vitamin C group, plasma vitamin C concentrations were significantly increased. Conversely, concentration of plasma vitamin C in Tocotrienol group increased from baseline of 2.72 +/- 0.20 mg/L to 6.80 +/- 0.63 mg/L and 8.9 +/- 0.77 mg/L respectively. Plasma concentrations of alpha-, delta- and gamma-tocotrienols in this group were significantly elevated. After 6-week washout period, all the elevated concentrations returned to basal levels. CONCLUSION: The study showed a good bioavailability of these vitamins and increment due to supplementation.

Suboptimal therapy and clinical management of gonorrhoea in an area with high-level antimicrobial resistance.

The choice of antimicrobial agents for the treatment of gonorrhoea is critical in areas where the prevalence of drug resistance is high. This study aimed to evaluate the antibiotic treatment of gonorrhoea in endemic areas. During 1999-2004, all Neisseria gonorrhoeae infections (n = 90) were evaluated. Patients' medical records and antibiotic treatment regimens were retrospectively reviewed if their isolates were viable (n = 65). In vitro antimicrobial susceptibility of N. gonorrhoeae isolates was performed. Urethritis (89%) and pelvic inflammatory disease (42%) were the most common presentations among men (n = 53) and women (n = 12), respectively. Of 54 patients with uncomplicated N. gonorrhoeae infection, 32 of them received appropriate antibiotics, including cefuroxime (n = 20), ceftriaxone (n = 10), ciprofloxacin (n = 1) and azithromycin (n = 1) during follow-ups. Among 65 patients, 53.8% were notified to the health authority. Check-ups of other sexually transmitted diseases were carried out in only 46% of patients. Not all isolates were susceptible to penicillin, 96.9% were resistant to tetracycline and 86% were resistant to ciprofloxacin. Ceftriaxone, cefixime, spectinomycin and azithromycin were active in vitro against all isolates. In conclusion, It is crucial to develop treatment guidelines according to regional antimicrobial resistances and educational programmes to improve clinical care for genital gonococcal diseases.

The prognostic influence of prevertebral space involvement in nasopharyngeal carcinoma.

OBJECTIVES: The purpose of this study was to evaluate the effect of prevetebral space involvement on treatment outcomes in patients with nasopharyngeal carcinoma (NPC) who were treated with radiotherapy/concurrent chemoradiotherpy or concurrent chemoradiotherpy with adjuvant chemotherapy. DESIGN: A retrospective review of case notes from the Kaohsiung Veterans General Hospital archives was performed. SETTING: A medical centre in Taiwan. PARTICIPANTS: There were 145 newly diagnosed cases of NPC. Thirty-nine patients were excluded because of the presence of distant metastasis at the time of presentation, loss of follow-up and incomplete image information. MAIN OUTCOME MEASURES: Pearson's chi-square tests were used to analyse correlation between tumour invasion and prevetebral space involvement during univariate analysis and logistic regression was applied during multivariate analysis. Kaplan-Meier survival curves were constructed. Multivariate analysis was performed to examine the impact of various prognostic factors. Pearson's chi-square and Fisher's exact test were also used to evaluate the correlation between failure patterns and treatment modality. RESULTS: A total of 106 patients with newly diagnosed NPC were enrolled in this study. Forty-three patients (41%) in this series were found to have prevertebral space involvement. Patients with prevertebral space involvement conferred a poor overall survival rate and metastasis-free survival rate compared with those without prevertebral space invasion (P = 0.04 and 0.02 respectively). Multivariate analysis showed that prevertebral space invasion was associated with an increased risk for distant metastasis [hazard ratio (HR) 14, 95% confidence interval (CI) 1.0-17.4; P = 0.03)] and overall survival (HR 7, 95% CI 1.1-135; P = 0.04). In patients with prevertebral space involvement, their metastasis-free survival rate, with and without adjuvant chemotherapy, was 100% and 72.7% (P = 0.047). This phenomenon was not observed in NPC patients without prevertebral space invasion. CONCLUSIONS: The present data revealed that prevertebral space involvement has a close relationship with survival rates and recurrence rates of patients with NPC. Nasopharyngeal carcinoma patients with prevertebral space involvement have more recurrence and poorer survival rates and should be the group to benefit from concurrent chemoradiotherapy followed by adjuvant chemotherapy. Inclusion of prevertebral space involvement may be needed to predict prognosis of NPC and help us to identify the high-risk group.

Branched projections in the auditory thalamocortical and corticocortical systems.

Branched axons (BAs) projecting to different areas of the brain can create multiple feature-specific maps or synchronize processing in remote targets. We examined the organization of BAs in the cat auditory forebrain using two sensitive retrograde tracers. In one set of experiments (n=4), the tracers were injected into different frequency-matched loci in the primary auditory area (AI) and the anterior auditory field (AAF). In the other set (n=4), we injected primary, non-primary, or limbic cortical areas. After mapped injections, percentages of double-labeled cells (PDLs) in the medial geniculate body (MGB) ranged from 1.4% (ventral division) to 2.8% (rostral pole). In both ipsilateral and contralateral areas AI and AAF, the average PDLs were <1%. In the unmapped cases, the MGB PDLs ranged from 0.6% (ventral division) after insular cortex injections to 6.7% (dorsal division) after temporal cortex injections. Cortical PDLs ranged from 0.1% (ipsilateral AI injections) to 3.7% in the second auditory cortical area (AII) (contralateral AII injections). PDLs within the smaller (minority) projection population were significantly higher than those in the overall population. About 2% of auditory forebrain projection cells have BAs and such cells are organized differently than those in the subcortical auditory system, where BAs can be far more numerous. Forebrain branched projections follow different organizational rules than their unbranched counterparts. Finally, the relatively larger proportion of visual and somatic sensory forebrain BAs suggests modality specific rules for BA organization.

The sulphorhodamine (SRB) assay and other approaches to testing plant extracts and derived compounds for activities related to reputed anticancer activity.

Since the major approach in searching for potential anticancer agents over the last 50 years has been based on selective cytotoxic effects on mammalian cancer cell lines, cell-based methods for cytotoxicity are described and compared. The sulphorhodamine B (SRB) assay is described in detail as the preferred method and also a novel approach has been developed which is based on the hypothesis that, in some circumstances, the naturally occurring compounds act as prodrugs rather than active compounds in their own right. Consequently, extracts or compounds are pre-incubated with systems modelling metabolic processes in the body before being tested. The methods have been validated using known compounds and Iris tectorum extracts have been shown to be more cytotoxic after treatment with beta-glucosidase. In addition bioassays based on mammalian cells involving antioxidant and upregulation of some cellular self-defence mechanisms are discussed which are related to prevention as well as treatment of cancer. Extracts of Alpinia officinarum induced glutathione-S-transferase (GST) activity in cultured hepatocytes and this was traced to the phenylpropanoids present, especially 1'-acetoxychavicol acetate.

Uses and abuses of in vitro tests in ethnopharmacology: visualizing an elephant.

Although in vivo models give a more accurate reflection of the activity of substances used in traditional medicine, their use in many countries is severely restricted due to economic and ethical concerns, and this has resulted in the widespread use of in vitro tests in ethnopharmacological studies. Such tests are very useful where the identity of compounds responsible for the biological activity of an extract is being investigated and where limited supplies of material are available, but it is important to consider a variety of factors before making over-predictive claims of that activity in one particular system explains the traditional use. The use of only one bioassay gives a very incomplete picture of the effect of the extract on the whole system involved. A symptom may be due to a number of disease states and, consequently, a variety of mechanisms may serve as targets for bioassays. In a similar way, it is very unusual for there to be only one target for a particular disease so a variety of test systems must be employed. Examples are given of batteries of test systems used to test plants and other materials with a reputation of being useful in wound-healing, diabetes, cancer and to treat cognitive decline associated with old age. In addition, consideration must be given to factors such as absorption into the body and metabolism of any substances present, either to decrease or increase the effect of the 'actives'.

Sheng mai san, a Chinese herbal medicine, protects against renal ischaemic injury during heat stroke in the rat.

1. There is evidence that the induction of inducible nitric oxide synthase (iNOS) and peroxynitrite by ischaemia/reperfusion may lead to renal cell injury. Herein, we investigated whether Sheng mai san (SMS), a Chinese herbal medicine, protects against renal ischaemic injury during heat stroke by reducing iNOS-dependent nitric oxide (NO) and peroxynitrite formation. 2. Urethane-anaesthetized rats were exposed to heat stress (ambient temperature 43 degrees C) to induce heat stroke. Control rats were exposed to 24 degrees C. Mean arterial pressure and renal blood flow after the onset of heat stroke were significantly lower in heat stroke rats than in control rats. However, both colonic temperature and renal damage score were greater in heat stroke rats compared with control rats. Similarly, plasma NO, creatinine and blood urea nitrogen (BUN), as well as the renal immunoreactivity of iNOS and peroxynitrite, were significantly higher in heat stroke rats compared with their normothermic controls. 3. Pretreatment with SMS (1.2 g/day per rat for 7 consecutive days before the initiation of heat stress) significantly attenuated the heat stroke-induced arterial hypotension, hyperthermia, renal ischaemia and damage, the increased renal immunoreactivity of iNOS and peroxynitrite and the increased plasma levels of NO, creatinine and BUN. Pretreatment with SMS resulted in a prolongation of survival time in heat stroke. 4. The results of the present study suggest that SMS protects against renal ischaemic damage by reducing iNOS-dependent NO and peroxynitrite production during heat stroke.

Cytotoxicity of plants from Malaysia and Thailand used traditionally to treat cancer.

The SRB cytotoxicity assay was used to screen extracts and isolated constituents of some traditional medicinal plants from Malaysia and Thailand against two human cancer cell lines, COR L23 lung cancer cell line and MCF7 breast cancer cell line and the non-cancer MCF5 cell line. Five out of the seven species tested, i.e. Thai Alpinia galanga, Alpinia officinarum, Cayratia japonica, Physalis minima, Tabernaemontana divaricata, exhibited interesting cytotoxicity activity and this is the first report of cytotoxicity from any Cayratia species. Following bioassay-guided fractionation, 1'-acetoxychavicol acetate (48h exposure against COR L23 cells, IC(50) 7.8 microM against MCF7 cells, IC(50) 23.9 microM) was isolated as the major cytotoxic component of the Alpinia species, physalin F as the major cytotoxic component of Physalis minima (48 h exposure against COR L23 cells IC(50) 0.4 microM against MCF7 cells, IC(50) 0.59 microM). The Malaysian Alpinia galanga showed weak activity compared with the Thai sample and this was shown to be due to the relatively high amounts of 1'-acetoxychavicol acetate present in the Thai sample.

Tonotopic and heterotopic projection systems in physiologically defined auditory cortex.

Combined physiological and connectional studies show significant non-topographic extrinsic projections to frequency-specific domains in the cat auditory cortex. These frequency-mismatched loci in the thalamus, ipsilateral cortex, and commissural system complement the predicted topographic and tonotopic projections. Two tonotopic areas, the primary auditory cortex (AI) and the anterior auditory field (AAF), were electrophysiologically characterized by their frequency organization. Next, either cholera toxin beta subunit or cholera toxin beta subunit gold conjugate was injected into frequency-matched locations in each area to reveal the projection pattern from the thalamus and cortex. Most retrograde labeling was found at tonotopically appropriate locations within a 1 mm-wide strip in the thalamus and a 2-3 mm-wide expanse of cortex (approximately 85%). However, approximately 13-30% of the neurons originated from frequency-mismatched locations far from their predicted positions in thalamic nuclei and cortical areas, respectively. We propose that these heterotopic projections satisfy at least three criteria that may be necessary to support the magnitude and character of plastic changes in physiological studies. First, they are found in the thalamus, ipsilateral and commissural cortex; since this reorganization could arise from any of these routes and may involve each, such projections ought to occur in them. Second, they originate from nuclei and areas with or without tonotopy; it is likely that plasticity is not exclusively shaped by spectral influences and not limited to cochleotopic regions. Finally, the projections are appropriate in magnitude and sign to plausibly support such rearrangements; given the rapidity of some aspects of plastic changes, they should be mediated by substantial existing connections. Alternative roles for these heterotopic projections are also considered.

Two novel mutations of the glycine receptor gene in a Taiwanese hyperekplexia family.

The authors report a Taiwanese family with autosomal recessive hyperekplexia. Two novel mutations, W96C (from the paternal allele) and R344X (from the maternal allele), which are located in exon 4 and exon 7 of the GLRA1 gene, were identified in this family. A series of electrophysiologic investigations were conducted in one of the probands, and the results suggest that the "startle center" is located subcortically.

Increased expression of alpha 1A Ca2+ channel currents arising from expanded trinucleotide repeats in spinocerebellar ataxia type 6.

The expansion of polyglutamine tracts encoded by CAG trinucleotide repeats is a common mutational mechanism in inherited neurodegenerative diseases. Spinocerebellar ataxia type 6 (SCA6), an autosomal dominant, progressive disease, arises from trinucleotide repeat expansions present in the coding region of CACNA1A (chromosome 19p13). This gene encodes alpha(1A), the principal subunit of P/Q-type Ca(2+) channels, which are abundant in the CNS, particularly in cerebellar Purkinje and granule neurons. We assayed ion channel function by introduction of human alpha(1A) cDNAs in human embryonic kidney 293 cells that stably coexpressed beta(1) and alpha(2)delta subunits. Immunocytochemical analysis showed a rise in intracellular and surface expression of alpha(1A) protein when CAG repeat lengths reached or exceeded the pathogenic range for SCA6. This gain at the protein level was not a consequence of changes in RNA stability, as indicated by Northern blot analysis. The electrophysiological behavior of alpha(1A) subunits containing expanded (EXP) numbers of CAG repeats (23, 27, and 72) was compared against that of wild-type subunits (WT) (4 and 11 repeats) using standard whole-cell patch-clamp recording conditions. The EXP alpha(1A) subunits yielded functional ion channels that supported inward Ca(2+) channel currents, with a sharp increase in P/Q Ca(2+) channel current density relative to WT. Our results showed that Ca(2+) channels from SCA6 patients display near-normal biophysical properties but increased current density attributable to elevated protein expression at the cell surface.

The C terminus of the Ca channel alpha1B subunit mediates selective inhibition by G-protein-coupled receptors.

Inhibition of calcium channels by G-protein-coupled receptors depends on the nature of the Galpha subunit, although the Gbetagamma complex is thought to be responsible for channel inhibition. Ca currents in hypothalamic neurons and N-type calcium channels expressed in HEK-293 cells showed robust inhibition by G(i)/G(o)-coupled galanin receptors (GalR1), but not by Gq-coupled galanin receptors (GalR2). However, deletions in the C terminus of alpha(1B-1) produced Ca channels that were inhibited after activation of both GalR1 and GalR2. Inhibition of protein kinase C (PKC) also revealed Ca current modulation by GalR2. Imaging studies using green fluorescent protein fusions of the C terminus of alpha(1B) demonstrated that activation of the GalR2 receptor caused translocation of the C terminus of alpha(1B-1) to the membrane and co-localization with Galphaq and PKC. Similar translocation was not seen with a C-terminal truncated splice variant, alpha(1B-2). Immunoprecipitation experiments demonstrated that Galphaq interacts directly with the C terminus of the alpha(1B) subunit. These results are consistent with a model in which local activation of PKC by channel-associated Galphaq blocks modulation of the channel by Gbetagamma released by Gq-coupled receptors.

Clinical characteristic of parenteral iron supplementation in hemodialysis patients receiving erythropoietin therapy.

BACKGROUND: Iron deficiency constitutes the major cause of erythropoietin hyporesponse in uremic patients receiving erythropoietin therapy; therefore, iron supplementation is necessary for these patients. Recent data suggested that intravenous iron supply is a preferable route for iron supplementation. However, it remains unclear whether a single large dose or multiple small doses are a better way of administering an intravenous iron supply. METHODS: To determine the effect of different dosing schedules of intravenous iron therapy on the hematocrit level, we randomly assigned 18 patients to 3 groups. The first group of patients (n = 6) received a single dose of 800 mg intravenous fesin (ferric saccharate). The second group of patients (n = 6) received 400 mg intravenous fesin once weekly for 2 successive weeks. The third group of patients (n = 6) received 120 mg of intravenous fesin for 7 successive hemodialysis sessions. EPO was given at a fixed dose for all individuals in the study period. RESULTS: The results showed that all 3 groups of patients had a progressive increase in hematocrit (Hct) level following intravenous iron therapy. Serum ferritin levels increased rapidly following iron therapy and then declined gradually in all 3 groups. But no statistical significance could be found among the 3 groups because of the small patient number. Also, no differences were observed in Hct or serum ferritin levels among these 3 groups of patients at all stages. CONCLUSION: In this study, we found that a large single dose as well as small multiple doses of parenteral iron therapy had similar effects in correcting the iron deficiency in hemodialysis patients treated with erythropoietin. To save manpower and costs, we recommend the large single dosing schedule.

T2-Weighted fast MR imaging with true FISP versus HASTE: comparative efficacy in the evaluation of normal fetal brain maturation.

OBJECTIVE: This study compares the relative efficacy of two fast T2-weighted MR imaging techniques-fast imaging with steady-state free precession (true FISP) and half-Fourier acquisition single-shot turbo spin-echo (HASTE)-in the evaluation of the normal fetal brain maturation during the second and third trimesters of gestation. SUBJECTS AND METHODS: The brain maturation of 10 normal nonsedated fetuses (5 during the second trimester and 6 during the third trimester of gestation [1 fetus underwent 2 examinations]) was examined by both techniques using a Vision+ 1.5-T MR system. We specifically looked for developing events, including white matter myelination, neuronal migration, and cortical sulcation. Image quality was graded according to the presence or absence of undesirable blurring. RESULTS: The specific absorption rate was lower for true FISP than for HASTE by a factor of 3 at equivalent imaging conditions. HASTE and true FISP provide comparable image quality in the second trimester when myelination of the cerebrum has not begun. Neuronal migration could be recognized as hypodense bands on both sequences during the second trimester. Myelination beginning at the third trimester was better delineated with true FISP than with HASTE because of point spread function-related blurring effects inherent in HASTE that hampered visualization of short-T2 structures. Cortical sulcation was well delineated by both sequences. CONCLUSION: With relatively superior image quality and significantly lower radiofrequency absorption than HASTE, true FISP is a safer and more effective alternative in the prenatal evaluation of normal fetal brain.

Poisoning by Datura leaves used as edible wild vegetables.

The causes of Datura intoxication include medication overdose, misuse of edible vegetables, deliberate abuse as a hallucinogen, homicidal or robbery and accidental intoxication from contaminated food. We report an incident of 14 people with Datura intoxication caused by ingesting wild Datura suaveolans for food. The incubation period was 15 to 30 min. The symptoms/signs were dizziness, dry mouth, flushed skin, palpitation, nausea, drowsiness, tachycardia, blurred vision, mydriasis, hyperthermia, disorientation, vomiting, agitation, delirium, urine retention, hypertension and coma. Three patients were hospitalized for 2-3 days. Thirteen persons received supportive fluid therapy. One patient did not receive medical therapy, he induced vomiting and drank a lot of water. Four patients presented with delirium/coma and 3 received physostigmine therapy with good response. One patient was intubated because of coma and respiratory depression. Three persons needed Foley catheterization for urine retention or coma status. One patient had a complication of urinary tract infection and antibiotic management. All patients recovered with no sequelae.

Noradrenergic neurotoxin suppresses gonadotropin-releasing hormone (GnRH) and GnRH receptor gene expression in ovariectomized and steroid-treated rats.

The present study was designed to investigate whether noradrenergic neurotransmission regulates the gene expression of gonadotropin-releasing hormone (GnRH) in the preoptic area and GnRH receptor in the pituitary. To this end, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4, 50 mg/kg), an intraperitoneal (i.p.) injection of selective noradrenergic neurotoxin, was administered 1 h before progesterone (1 mg) treatment in ovariectomized and estradiol-treated prepubertal rats. Treatment with DSP4 effectively blocked the progesterone-induced increase in hypothalamic noradrenaline content, but not dopamine content, indicating that DSP4 selectively inhibits noradrenergic neurotransmission. DSP4 significantly blocked progesterone-induced increase in serum luteinizing hormone (LH) concentrations as well as GnRH release from hypothalamic fragments incubated in vitro. DSP4 concomitantly down-regulated GnRH mRNA levels in the preoptic area, as determined by competitive reverse transcription-polymerase chain reaction. DSP4 also clearly down-regulated progesterone-induced GnRH receptor mRNA levels in the pituitary, whereas it failed to alter LHbeta mRNA levels. In summary, blockade of noradrenergic neurotransmission with DSP4 resulted in profound reductions of hypothalamic GnRH and pituitary GnRH receptor gene expression.

Isolation, characterization, and chromosomal mapping of a novel cDNA clone encoding human selenium binding protein.

We have isolated the full-length human 56 kDa selenium binding protein (hSP56) cDNA clone, which is the human homolog of mouse 56 kDa selenium binding protein. The cDNA is 1,668 bp long and has an open reading frame encoding 472 amino acids. The calculated molecular weight is 52.25 kDa and the estimated isoelectric point is 6.13. Using Northern blot hybridization, we found that this 56 kDa selenium binding protein is expressed in mouse heart with an intermediate level between those found in liver/lung/kidney and intestine. We have also successfully expressed hSP56 in Escherichia coli using the expression vector-pAED4. The hSP56 gene is located at human chromosome 1q21-22).