Specific PCR assays for the detection and quantification of DNA from the biocontrol strain Trichoderma harzianum 2413 in soil.

Strain identification in situ is an important factor in the monitoring of microorganisms used in the field. In this study, we demonstrated the use of sequence-characterized amplified region (SCAR) markers to detect genomic DNA from Trichoderma harzianum 2413 from soil. Two primers (SCAR A1/SCAR A1c) were tested against DNA of 27 isolates of Trichoderma spp. and amplified a 990-bp fragment from T. atroviride 11 and a 1.5-kb fragment from T. harzianum 2413, using an annealing temperature of 68 degrees C. These fragments showed no significant homology to any sequence deposited in the databases. The primer pair, BR1 and BR2, was designed to the 1.5-kb fragment amplified from T. harzianum 2413, generating a SCAR marker. To test the specificity of these primers, experiments were conducted using the DNA from 27 Trichoderma spp. strains and 22 field soil samples obtained from four different countries. PCR results showed that BR1 and BR2 amplified an 837-bp fragment unique to T. harzianum 2413. Assays in which total DNA was extracted from sterile and nonsterile soil samples, inoculated with spore or mycelium combinations of Trichoderma spp. strains, indicated that the BR1 and BR2 primers could specifically detect T. harzianum 2413 in a pool of mixed DNA. No other soil-microorganisms containing these sequences were amplified using these primers. To test whether the 837-bp SCAR marker of T. harzianum 2413 could be used in real-time PCR experiments, new primers (Q2413f and Q2413r) conjugated with a TaqMan fluorogenic probe were designed. Real-time PCR assays were applied using DNA from sterile and nonsterile soil samples inoculated with a known quantity of spores of Trichoderma spp. strains.

[Therapy of postmenopausal osteoporosis]. / Therapie der postmenopausalen Osteoporose.

The therapy of postmenopausal osteoporosis is based on a few comprehensible assumptions. High bone resorption should be reduced by treatment with bisphosphonates, raloxifene or seldom with calcitonins. After reduction of high bone turnover and in low bone turnover situations, an osteoinductive combination therapy should be started, inducing collagen type I with parathyroid hormone or fluorides. This collagen can then be mineralized by calcium, vitamin D, and vitamin D metabolites. In addition, bone resorption should be reduced during menopause with estrogens and gestagens, in the case of a receptor-positive breast cancer with tamoxifen, and after menopause with raloxifene or a bisphosphonate. In elderly patients a depletion of vitamin D often induces an osteoporomalacia instead of an osteoporosis. In this situation, mineralization of the osteoid by calcium and vitamin D is sufficient for therapy. A daily osteoporosis gymnastic program is required and physical activity should be enhanced to increase muscle mass because bone adapts to the individual situation.

[Differences in the psychological status of hysterectomy and non-hysterectomy women]. / Unterschiede in der psychischen Befindlichkeit zwischen hysterektomierten und nicht-hysterektomierten Frauen.

PURPOSE OF THE STUDY: Despite numerous investigations on the well-being of hysterectomized women, this issue is still discussed controversially. The aim of the present study was to reveal differences between a group of hysterectomized women with a fairly long follow-up period (7.9 years) and a group of non-hysterectomized women with respect to their psychological well-being. RESEARCH METHOD AND MATERIAL: In a questionnaire study we compared a group of 216 women who underwent hysterectomy with a group of 90 non-hysterectomized women. Dependent variables were: body complaints, psychological well-being, dysphoria, socio-sexual assertiveness, and gender-role orientation. RESULTS AND CONCLUSIONS: The group of hysterectomized women showed significantly higher levels of body complaints, depression, and unassertiveness in sexual situations, as well as decreased psychological well-being, and a more traditional gender-role orientation. Within the group of hysterectomized women, duration since hysterectomy, surgical techniques, and hormone substitution therapy seem to be unrelated to the psychological variables. The group differences cannot be causally attributed to the hysterectomy, since they may already have existed premorbidly. Nevertheless, the results suggest that a more thorough psychological examination prior to hysterectomy, as well as provision of support for the coping process after intervention is advantageous.

cAMP-synthesis in a medullary thyroid carcinoma cell line: response to adrenergic agents and prostaglandines.

Calcitonin secretion by C-cells is mediated through intracellular 3'5'-cyclic adenosine monophosphate (cAMP) and calcium signaling. Calcitonin release stimulation tests may take advantage of both signaling cascades in screening for medullary thyroid carcinomas (MTC). To elucidate the regulation of the adenylyl cyclase system we have determined cAMP levels of a calcitonin-expressing MTC cell line (RG) after exposure to adrenergic agents and prostaglandines. In early passages (20-30) cAMP concentrations were significantly elevated in RG cells after exposure to beta-adrenergic agents and prostaglandines E1 and E2. In advanced passages (60-80) the beta-adrenergic response was no longer detectable and adrenergic receptors were uncoupled from the adenylyl cyclase complex; while the effect of prostaglandines E1 and E2 remained unaffected. Preincubation with dexamethasone, in a process requiring protein new synthesis, re-established the adrenergic response in later passages, indicating that RG cells dedifferentiated in culture over time. Our in vitro findings suggest that MTC cell dedifferentiation may be accompanied by adrenergic receptor-uncoupling from the adenylate cyclase system and that this process may be reversed by dexamethasone incubation.

PLENA and FARINELLI: redundancy and regulatory interactions between two Antirrhinum MADS-box factors controlling flower development.

We report the discovery of an Antirrhinum MADS-box gene, FARINELLI (FAR), and the isolation of far mutants by a reverse genetic screen. Despite striking similarities between FAR and the class C MADS-box gene PLENA (PLE), the phenotypes of their respective mutants are dramatically different. Unlike ple mutants, which show homeotic conversion of reproductive organs to perianth organs and a loss of floral determinacy, far mutants have normal flowers which are partially male-sterile. Expression studies of PLE and FAR, in wild-type and mutant backgrounds, show complex interactions between the two genes. Double mutant analysis reveals an unexpected, redundant negative control over the B-function MADS-box genes. This feature of the two Antirrhinum C-function-like genes is markedly different from the control of the inner boundary of the B-function expression domain in Arabidopsis, and we propose and discuss a model to account for these differences. The difference in phenotypes of mutants in two highly related genes illustrates the importance of the position within the regulatory network in determining gene function.

alpha-Tocopherol supplementation in racing cyclists during extreme endurance training.

This study was undertaken to evaluate the effects of 5 months of alpha-tocopherol supplementation on physical performance during aerobic exercise training in 30 top-class cyclists. Antioxidative effects of supplementation were also studied. Plasma alpha-tocopherol concentration increased significantly in the vitamin E-supplemented group, whereas the placebo group showed a trend toward decrease. Physical performance did not improve in the alpha-tocopherol-supplemented group compared to the placebo group. Heart rates were also not significantly different. Lactate concentrations at the aerobic threshold and the anaerobic threshold were identical. Thus, there was no performance improvement in the alpha-tocopherol-supplemented group. However there was a significant reduction in CK in serum of the E-supplemented group. A trend toward decrease in GOT, GPT, and LDH was observed with alpha-tocopherol supplementation. Moreover, significantly reduced malondialdehyde serum levels were measured in the E-supplemented group. The findings indicate a protective effect of alpha-tocopherol supplementation against oxidative stress induced by strenuous exercise.

Therapy of steroid-induced bone loss in adult asthmatics with calcium, vitamin D, and a diphosphonate.

In a prospective, controlled, and randomized clinical trial, we examined the effects of treatment with vitamin D (1,000 IU/d), calcium (1 g/d), and ethane-1-hydroxy-1,1-diphosphonate (EHDP; 7.5 mg/kg body weight) on vertebral bone mass in fourteen asthmatics undergoing long-term treatment with systemically applied corticosteroids. The extent of steroid-induced bone loss was judged by vertebral bone density of the lumbar spine measured by dual-photon absorptiometry as well as by vertebral crush fracture incidence examined by conventional X-ray. Results of the measurements before treatment and after six mo were compared with those of an untreated control group of nineteen asthmatics. Bone density increased during the observation period by 5% in the treated group, compared with a decrease of 4.3% in the untreated control group (p < 0.01). Moreover, in the treated group no radiologically visible new fractures occurred; in the control group new fractures were observed in four patients. There were no serious side effects of the applied drugs during the 6-mo period. Therefore, the combination of EHDP, calcium, and vitamin D appears to be a useful regimen for the management of steroid-induced bone loss in adult asthmatics.

Assessment of vitamin B2 status in performance athletes of various types of sports.

To determine the vitamin B2 supply, 62 athletes in various types of sports and 16 non-athletes were examined, using a weighed food record (7-day), alpha-EGR determination, microbiological blood, and HPLC urinary measurements. If the guidelines of 1.5 or 1.7 mg/day vitamin B2 or 0.6 mg/4,184 kJ (GRDA) are defined for adequate status, all sports groups (M +/- SEM) are above this level. The vitamin B2 blood concentrations show a range (mean) from 303-372 nmol/liter, which indicates that more than 90% of the athletes are above the reference value of 327 nmol/liter (mean). There is a correlation of r = 0.49 between the vitamin B2 intake (mean over 7 days) and the vitamin B2 blood level. Determination of vitamin B2 blood concentration is characterized by good precision (C.V. +/- 3.2%) and recovery (95.7%). The enzyme activation measurement (alpha-EGR) shows that all of the athletes are below the alpha-EGR reference value of < 1.50 (good supply). No significant correlations could be determined between alpha-EGR and vitamin B2 intake, blood concentrations, and urinary excretion. The measurement also shows high precision (C.V. 1.7%). The Vitamin B2 excretion in urine (mumol/g Cr.) was higher than the reference values for non-athletes in more than 90% of the athletes. There is a correlation of r = 0.89 between the vitamin B2 intake on the last day (Day 7) of the food record and the riboflavin excretion, and between vitamin B2 in blood and urinary excretion of r = 0.52. The C.V. of vitamin B2 urinary measurement using HPLC is 2.5%, with recovery of 98.2%. Combined measurement of vitamin B2 intake, blood concentration, and urine excretion appears to be a suitable procedure for determining the vitamin B2 status. Most of the performance athletes were sufficiently supplied with vitamin B2, compared to the reference values for non-athletes.

Serum osteocalcin and vitamin D metabolites in patients with ankylosing spondylitis.

OBJECTIVES: Osteocalcin is the major non-collagenous protein of bone and is regarded as a specific index of bone formation. The aim of this study was to examine the rate of bone formation measured by osteocalcin in 38 patients with ankylosing spondylitis (AS) and its dependence on various parameters of calcium and phosphate metabolism. METHODS: Serum osteocalcin, alkaline phosphatase, parathyroid hormone, and 1,25-dihydroxyvitamin D were measured in 38 patients with ankylosing spondylitis and in 52 controls. RESULTS: Mean serum osteocalcin was significantly reduced in patients with AS (men 1.7 (1.1) ng/ml; women 1.2 (1.1) ng/ml) compared with the corresponding control groups (men 3.2 (1.3) ng/ml; women 4.1 (1.7) ng/ml). In contrast, alkaline phosphatase was only slightly but significantly higher (135 (44) U/l) in patients with AS than in the corresponding controls (114 (35) U/l). Serum parathyroid hormone (AS 3.1 (0.7) v 2.7 (0.6) mE/ml) and 1,25-dihydroxyvitamin D (AS 64.0 (34.5) v 52.4 (6.7) pg/ml) were slightly but not significantly higher in patients with AS. Consequently, as both hormones are known to stimulate osteocalcin synthesis, they are not responsible for low osteocalcin levels in patients with AS. No significant correlation between alkaline phosphatase and osteocalcin was found. Low serum levels of osteocalcin in patients with AS reflect lower osteoblastic activity in AS. CONCLUSIONS: Bone turnover in patients with AS is characterised by low bone formation in the presence of normal levels of calcium regulating hormones.

[Closure of a persistent ductus arteriosus Botallo without thoracotomy]. / Verschluss des persistierenden Ductus arteriosus Botalli ohne Thorakotomie.

A persisting ductus arteriosus (PDA) was closed with the Rashkind occlusion system in 15 patients (12 children, aged 1.0 to 10.8 years, and three adults, aged 21, 33 and 56 years). The method consists of the implantation of a double umbrella of polyurethane foam mounted on platinum or steel wire introduced into the PDA via a transport catheter, the ductus being closed by thrombosing and epithelialization of the implant. The mean diameter of the PDA (as measured in the lateral aortogram) was 2.4 (1.4-5.3) mm. There were no complications of the procedure, nor any embolization or other serious complications. The PDA was completely occluded in eight patients, while a residual shunt remained in seven. In the oldest patient, aged 56 years, who had a residual shunt, increasing haemolysis required surgical closure of the ductus with a pericardial patch. These results indicate that, for patients weighing over 5 kg and with a ductus diameter of less than 10 mm, the closure method after Rashkind is a sparing and only slightly invasive procedure.

[Successful operation of right ventricular cardiac fibroma in an infant]. / Erfolgreiche Operation eines rechtsventrikulären kardialen Fibroms bei einem Säugling.

Cardiac tumors are rare in infants. We describe the diagnostic procedure and the good postoperative outcome despite the very large cardiac fibroma.

[Fracture incidence in 803 women with postmenopausal osteoporosis under drug treatment]. / Frakturinzidenz unter der medikamentösen Behandlung bei 803 Frauen mit einer Postmenopause-Osteoporose.

In a survey of cases of confirmed postmenopausal osteoporosis, 803 women were asked to complete a questionnaire on radiologically confirmed fractures sustained prior to and during drug therapy. Under monotherapy (fluoride or calcium, n = 92) the number of peripheral and central fractures was 54% and 49% lower, respectively. The corresponding figures with two substances (fluoride and calcium, n = 226) were 58% and 60%, with three substances (fluoride, calcium and estrogens or Vitamin D, n = 223) 83% and 72%, and with four substances (n = 254) 85% and 75%.

Hemodynamic findings during exercise on a bicycle ergometer following balloon valvuloplasty of pulmonary stenosis in children and adolescents.

Eleven patients (4 female, 7 male), age range 3.3 to 24.8 years (mean 11.10 years) treated for isolated pulmonary stenosis underwent cardiac catheterization and percutaneous transluminal balloon valvuloplasty (PTVP). The right ventricular systolic pressure (RVSP) before valvuloplasty ranged from 31 to 127 mmHg (mean 79 mmHg) decreasing to 28 to 62 mmHg (mean 42 mmHg) immediately after the dilatation. The peak systolic gradient of the pulmonary valve (delta p RV-PA) before valvuloplasty ranged from 22 to 107 mmHg (mean 61 mmHg) and decreased to a range of 14 and 45 mmHg (mean 23 mmHg) immediately after the dilatation. Balloon valvuloplasty was performed using balloons of 13 to 31 mm in diameter. On 11 patients cardiac catheterization and Doppler echocardiography were repeated between 11 months and 5.3 years (mean 3.11 years) after the balloon valvuloplasty showed a further significant fall in the gradient of pressure. The right ventricular systolic pressure ranged from 20 to 51 mmHg (mean 31.7 mmHg) while the transpulmonary gradient varied from 3 to 24 mmHg (mean 11.6 mmHg). At the time of follow-up examination the patients were aged between 7.2 and 25.7 years (mean 15.9 years). On average the second catheterization was performed 3.11 years following the first hemodynamic study. The follow-up examination encompassed clinical examination, electrocardiogram, Doppler echocardiography, and right heart cardiac catheterization. During right heart cardiac catheterization the children exercised on a bicycle ergometer for three min at 50 or 100 W depending on their body surface area. During this exertion, pressures of the right ventricle and the pulmonary artery as well as heart rate and oxygen saturation were recorded.(ABSTRACT TRUNCATED AT 250 WORDS)

Improved neurological outcome following early anatomical correction of transposition of the great arteries.

The incidence of neurological residuals following anatomical correction of transposition of the great arteries (d-TGA) has not been described so far. Clinical examination, EEG recordings, and computed tomography (CT) scans were carried out in a consecutive series of 38 children with d-TGA surviving anatomic corrective surgery. The patients were classified into one of three groups according to the type of operation: 15 patients after two-stage approach (TSA) (Stage 1: pulmonary artery banding+aortopulmonary shunt; Stage 2: anatomic correction); 12 patients with primary anatomic correction within the first 2 weeks of life (early switch, ES); 11 patients with primary anatomic correction later in infancy (later switch, LS). In 26 patients (68%) we found no abnormalities on neurologic examination, CT scan, or EEG. Four patients suffered from spastic hemiplegia, 3 of these had cortical brain damage visible on CT scan, and 3 had focal epilepsy as well. In 2 otherwise clinical normal patients cortical infarction could be seen on a CT scan. Thus, in 5 cases (13% of 38 patients) cerebral infarcts were diagnosed by CT scan. The cortical vascular infarction was seen in 4 patients after TSA and in 1 after LS. In 6 patients we found other neurological abnormalities. Early anatomic correction in patients with d-TGA reduces the risk of cortical vascular infarction.

Measurement of ascorbic acid and erythorbic acid in processed meat by HPLC.

A simple and efficient HPLC method is described to measure both ascorbic and erythorbic acid separately in processed meat products. The system used consisted of a reversed-phase column, a mobile phase containing an ion-pairing agent and ultraviolet detection. The procedure, which was checked on a variety of samples, was found to give reproducible and reliable results.

Quantification of biomineralization: an in-vitro tissue culture system and microanalysis of calcium, phosphorus and trace elements by total-reflection X-ray fluorescence.

An in-vitro tissue culture system with folded periostea of 17-day-old fetal chick calvaria was combined with analytical methods to achieve quantification of biomineralization. A scanning electron microscope with an energy-dispersive X-ray detector was applied to show the distribution of calcium, phosphorus and trace elements. Calcium and phosphorus were concentrated in the zone of the mineralized matrix. Strontium was distributed similar to calcium. Zinc was distributed equally in the soft tissue and the mineralized matrix. Total-reflection X-ray fluorescence was used for quantification. Thyroxine in high concentration reduces the calcium content of the samples. One week after incubation magnesiumchloride (1.8 mM) or zincchloride (.1 mM) were found to reduce the calcium content by 38% or 82%, respectively.