[Clinical microbiology as a basis of antimicrobial chemotherapy].

Clinical microbiology is a facet of medical microbiology which is dedicated to all aspects of human infectious disease diagnosis, especially in patients treated in hospitals. The specific tasks of such a laboratory can be solved at best, if it is integrated in the hospital and if there is close co-operation with clinical specialists. Different methods for susceptibility testing of isolated bacteria are possible: agar diffusion test, agar dilution test, bouillon dilution test etc., the results of which need to be interpreted considering further aspects. During antimicrobial chemotherapy certain tests are also necessary or useful. The objective of future developments is to avoid the need of cultures and to establish antibiotic profiles within hours to provide the clinician with relevant results.


[Antibiotic therapy of bacterial respiratory tract infections].

Infections of the airways are among the most frequent of inflammatory diseases appropriate for antibacterial therapy. One must differentiate between infections of the upper and lower airways, respectively infections obtained externally, and nosocomial infections. Since a relatively uniform spectrum of microbes can usually be expected, well-planned antibacterial therapy is possible. For nosocomial infections, on-target antibacterial therapy is required. For the various diseases of the airways, diagnostic and therapeutic strategies are outlined.


[Diagnosis and therapy of urinary tract infections].

Urinary tract infections (UTI) are one of the most frequent indications for antibacterial chemotherapy. We must be differentiate between complicated and uncomplicated as well as between upper and lower UTI. Regarding the definition of significant bacteriuria (> or = 10(5) CFU/ml), some relativations are necessary. Dip-slide techniques proved to be very useful for bacteriological diagnosis. The calculated therapy depends upon age and sex and is different for community-acquired and nosocomial infections. Further items are the selection of antibacterial agents, the duration of chemotherapy, the role of asymptomatic bacteriuria and reinfection prophylaxis.


[Antimicrobial chemotherapy of infection in intensive care conditions].

Antimicrobiologic chemotherapy is a cornerstone in the modern concept of treatment of sepsis. It is supported by a number of measures of intensive care. Externally acquired infections followed by sepsis mainly affect immunocompromised patients. They represent the minority. On the other hand, nosocomial infections play a dominant role in intensive care units. Nosocomial pneumonias and infections caused by intravascular plastics or endoprotheses are the most important reasons of sepsis. Initial antimicrobiologic therapy considers both the infectious focus and the specific epidemiology and resistance of microorganisms present in the department: it comprises the drawing of 2 to 3 blood cultures, taking of urinary cultures, tracheal secretion, liquor and wound-swaps for microbiologic examination. A gramstaining of the preparation can be helpful. In case the focus of the sepsis is not known, the first step of treatment consists of a combination of piperacilline or cephalosporins with aminoglycosides. If the septic state does not improve within 48 hours, a glycopeptide should be added against staphylococci (second step). If the combination fails, imipenem/cilastatin and aminoglycosides are administered as the third step. Normally, the result of cultures and resistogram already are available by this time. Today aminoglycosides are preferably given once daily, the serum level is monitored, and the toxicity of aminoglycosides is thus diminished.


[Therapy of bacterial infections in neutropenic and immunocompromised patients].

Neutropenias, especially extended an long-lasting stages, lead to life-threatening endogenous infection. Therefore, after taking off materials for bacteriological investigations an empirical schedule of a combined high dose, treatment with broad-band antibiotics and/or antimycotics has immediately to be introduced and to continue until the body temperature and the peripheral blood granulocytes are normalized. In case of treatment failure one should complete the therapy by other additional antibiotics or correct the combination of its in respect to the results of the microbiological investigations. Supplements of this antimicrobial treatments are immunoglobulins and growth factors (G-CSF, GM-CSF). In case of an expected neutropenica the use of the selective gut decontamination or the reverse isolation of the patient can be of essential advantage.


[Fungal infections in granulocytopenic and immunocompromised patients].

Opportunistic fungus infections in neutropenic immunocompromised patients have strikingly increased, especially with the improvement of antibiotic treatment. Their outcome is often fatal because of the difficulties in diagnosis and treatment. Therefore a rationale of surveillance diagnostics and empiric treatment in risk patients is necessary. In these patients a continuous weekly mycotic diagnosis of mouth, throat, faeces, urine, vagina, as well as of the blood is necessary. During an aggressive neutropenia-producing chemotherapy an antimycotic prophylaxis with the aim of reducing fungal colonization in the gastrointestinal tract (sometimes in the respiratory pathways, too) should be performed. Fever of unknown origin lasting longer than 4-5 days in spite of broad spectrum antibiotic treatment and/or positive diagnostic findings must lead to treating risk patients empirically using amphotericin B 1 mg/kg/d or a combination of amphotericin B 0.3-0.5 mg/kg/d together with flucytosin (Ancotil) 150 mg/kg/d. In case of a beginning candidiasis, patients can first be treated with fluconazol (Diflucan). The dose is 400 mg/d, later on 200 mg/d. It is pointed out that, much more often than usual, in risk patients with fever, atypical pneumonia, meningoencephalitis or other organ symptoms fungal infections should be taken into consideration. The most common opportunistic fungal diseases are presented and details concerning the different antimycotic drugs are given.


[The value of ultrasound and computerized tomography in detection of cystic changes in chronic pancreatitis].

Intra- or extrapancreatic pseudocysts (PP) are the most common local complication in chronic pancreatitis. Aim of this study was to investigate frequency, localisation and size of pseudocysts in patients with chronic pancreatitis by means of ultrasound (US) and computed tomography (CT). 155 patients (females 35, males 120) with chronic pancreatitis, that underwent simultaneous (within two weeks) CT and US examinations, from January 1982 to June 1989, were included in this study. Cystic lesions were detected in 62% by CT, in 52% by US. Sensitivity in detection of cysts based on intraoperative findings (gold standard) was 98% for CT and 94% for US. 80% of the pseudocysts were smaller than 6 cm. 46% were in the range from 2 to 66 cm and 34% were smaller than 2 cm. The most common localisation was the pancreatic head region (50%), 20 of 102 patients with chronic pancreatitis were found to have a direct communication of a pseudocyst with the ductal system by ERP. No specific clinical or laboratory pattern were associated with the presence of pseudocysts. Increased pancreatic serum amylase concentration was detected in 29% of patients with and in 27% of patients without pseudocysts.


[Results of conventional and laparoscopic cholecystectomy].

We report about a retrospective study of 861 conventional and 812 laparoscopic cholecystectomies (including one coelioscopic choledocholithotomy). In the conventionally operated group reoperation was required in 2.7% (1.4% relaparotomy, 1.3% secondary suture), mortality was 0.5%. After laparoscopic treatment the reoperation rate was 2% (10 relaparoscopies, 5 laparotomies, 2 secondary sutures), no mortality. Growing experience and better definition of contraindications for endoscopic cholecystectomy might improve our results in the future.


Enteric protein loss as a marker of intestinal inflammatory activity in Crohn's disease: comparability of enteric clearance and stool concentration of alpha-1-antitrypsin?

Intestinal alpha 1-antitrypsin (alpha 1-AT) clearance has been shown a reliable index of intestinal inflammatory activity in Crohn's disease (CD). For reasons of practicability, it has been repeatedly suggested to replace alpha 1-AT clearance by alpha 1-AT concentration in random stool samples. In 60 controls and in 70 patients with CD, in 21 patients before and after treatment, fecal alpha 1-AT concentration and the ratio of stool and serum alpha 1-AT concentration were compared with alpha 1-AT clearance. In 11 patients alpha 1-AT clearance, fecal concentration and stool/serum alpha 1-AT concentration ratio were compared with 51Cr-albumin clearance. alpha 1-AT clearance (104 +/- 14 vs. 17.5 +/- 2 ml/d, p < 0.0001) as well as fecal alpha 1-AT concentration (155 +/- 21 vs. 30 +/- 3 mg/100 ml, p < 0.0001) and stool/serum alpha 1-AT concentration ratio (45 +/- 6 vs. 12 +/- 1) were significantly higher in CD patients than in controls. alpha 1-AT clearance (60 +/- 9 vs. 37 +/- 4 ml/d, p < 0.01), fecal alpha 1-AT concentration (113 +/- 21 vs. 59 +/- 8 mg/100 ml, p < 0.01) and the stool/serum alpha 1-AT concentration ratio (27 +/- 4 vs. 18 +/- 2) decreased after treatment, but fecal alpha 1-AT concentration and the stool/serum alpha 1-AT concentration ratio failed to parallel the course of alpha 1-AT clearance in 33% and in 24% of patients, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


[Therapy of stomach ulcer--a comparison between the low dosage antacid hydrotalcite and ranitidine--results of a randomized multicenter double-blind study. Talcivent Study Group].

In a 8 week double-blind randomized multicenter trial in 159 patients with benign gastric ulcer the efficacy of hydrotalcite vs. ranitidine in expediting ulcer healing and in achieving pain relief was determined. 79 patients received hydrotalcite 1000 mg q.i.d. as tablets equalling a total neutralizing capacity of 111.2 mval and 80 patients received ranitidine (300 mg at night). Endoscopically controlled healing rates after 4 weeks of therapy amounted to 41.8% with hydrotalcite and 53.8% with ranitidine. After 8 weeks both regimen showed significant equivalent healing rates (hydrotalcite: 81.0%, ranitidine: 78.8%, p < 0.003). Ulcer pain decreased parallel in both groups. By the end of therapy 92.4% of the patients treated with hydrotalcite and 86.3% of those receiving ranitidine were free of pain. Incidence of helicobacter pylori in antral mucosal biopsies was not influenced by both treatments. We conclude that an 8-week treatment with low dose hydrotalcite therapy is as effective as ranitidine in healing benign gastric ulcers and achieving pain relief.


[Diffuse cholangiofibromatosis--incidental findings in sonography, ERCP and laparoscopy].

Diffuse cholangiofibromatosis is a rare and asymptomatic abnormality. Its clinical significance results from problems that occur in differential diagnosis to granulomatous hepatitis, sclerosing cholangitis, multiple microabscesses or diffuse tumour infiltration of the liver, when it is incidentally found in ultrasound, ERCP or laparoscopy. The characteristic findings in these examinations are described in two case reports. The ultrasonic pattern of multiple intrahepatic double structures, microcystic areas and surrounding hyperechoic reflexes are in accordance with multiple intrahepatic microcystic and hazy areas in ERCP x-rays and multiple white-yellowish areas that retract liver surface in laparoscopy. Final diagnosis is confined by the characteristical microscopic finding of von Meyenburg-complexes in liver biopsy.


Active autoimmune gastritis without total atrophy of the glands.

To date, autoimmune gastritis has been diagnosed for the most part only when total atrophy of the oxyntic glands is detected. On the basis of 40 patients without total atrophy of the glands, and with parietal cell antibodies in the serum, we show that the diagnosis of type A gastritis is also possible in the pre-atrophic stage. The histological criteria for the diagnosis of active autoimmune gastritis without total atrophy of the glands are 1. usually dense, diffuse locally emphasized lymphocytic infiltration of the lamina propria between the glands in the oxyntic mucosa, 2. focal destruction of individual glands in the corpus of the stomach by lymphocytes, and 3. reactive pseudohypertrophy of the parietal cells. A comparison with a group of patients with autoimmune gastritis and total atrophy of the glands shows that in active autoimmune gastritis, too, women are more frequently affected than men (in both groups, the sex ratio is approximately 3:1). Patients without atrophy of the glands are, on average, about 12 years younger than those with "burnt out" type A gastritis (average age 69.98:57.80 years). While in the case of burnt out type A gastritis, no colonisation with Helicobacter pylori was to be found, such colonisation was demonstrated for the corpus mucosa in 22.5%, and for the antral mucosa in 15.0%. In 27.5% a minimal or low-grade inactive superficial gastritis, as may be seen after eradication of Helicobacter pylori, was additionally diagnosed in the antrum. A knowledge of the histological appearance of the pre-atrophic stage of type A gastritis might be of importance for the possible prevention of pernicious anaemia.(ABSTRACT TRUNCATED AT 250 WORDS)


[Treatment of cholestatic liver diseases; the role of ursodeoxycholic acid].

Ursodeoxycholic acid (UDCA) allows symptomatic treatment of cholestatic liver diseases such as primary biliary cirrhosis, primary sclerosing cholangitis, intrahepatic biliary atresia, and cholestasis of cystic fibrosis. Patients should be treated at an early stage of the disease in order to prevent progression to cirrhosis. Since UDCA has no toxic effects longterm treatment with this substance is possible without the risk of undesired side effects. In patients with primary biliary cirrhosis and rapid progression of the disease, UDCA may be combined with an immunosuppressive substance (i.e. cyclosporin). In primary sclerosing cholangitis, biliary atresia and cholestasis of cystic fibrosis, UDCA at present seems the only treatment of which a benefit for the patients can be expected. In endstage disease liver transplantation is indicated. The role of UDCA in chronic hepatitis and alcohol induced liver disease needs to be clarified in further studies. Whether the improvement of laboratory tests in such patients indicates amelioration of the course of disease, still is unclear.


[Percutaneous transhepatic cholangioscopic color laser lithotripsy in bile duct calculi].

Tunable dye laser lithotripsy is an effective and low risk treatment in patients with bile duct stones in which transpapillary maneuvers failed. The percutaneous approach allows to introduce small caliber endoscopes (10.5 F) to fragment the calculi under vision. This technique was evaluated in 8 patients who had undergone a biliodigestive anastomosis or in whom the biliary calculi could not removed by standard retrograde treatment. Laser lithotripsy resulted in sufficient fragmentation in 7 patients. Bile duct clearance proved to be a particular problem with the percutaneous access. When a retrograde sphincterotomy is not possible an antegrade papillotomy must be attempted under fluoroscopic guidance. In bile duct strictures the implantation of expandable stents facilitates the passage of fragments and may prevent recurrent stricture and development of new stones.


[Computerized tomography analysis of gallstones: soon an essential diagnosis before nonsurgical gallstone therapy?].

Successful application of non operative procedures for gallstone therapy are limited on cholesterol stones. Therefore reliable therapeutic analysis of stone components is mandatory for planning of an adequate therapy. Computed tomographic gallstone analysis is considered to improve selection of patients significantly. However, standardized conditions for investigation of patients are not yet established. It is suggested that computerized tomography should be performed using 4 mm slides, standardized position of patients and a phantom, which allows calibration of different CT-machines.


[Psychoimmunologic correlation between allergies, panic and agoraphobia].

The present behavioral medical paper is a contribution to basic research in neuroimmunology. The relationship of panic/agoraphobia and allergy/anaphylaxis (IgE-mediated immediate reaction) is demonstrated and discussed with specific consideration of vasomotor reactions, tachycardia and hyperventilation in panic patients and allergies with anaphylactic reactions. The vegetative symptoms of panic reactions and anaphylaxis showed a high correlative correspondence. 74% of 23 panic and/or agoraphobia patients in a pretest to an epidemiologic study turned out to have an allergic illness that needed treatment; but there were no more anxiety disorders disclosed in 50 allergic patients than in the general population according to expectancies found in epidemiologic publications. If these data are confirmed then panic and agoraphobia cannot be seen as merely a cognitive-emotional event, but also as a further kind of allergic disease with genetic, ecological, psychological and social determinants; they would then have to be diagnosed as psychoimmunological disturbances and treated with behavior medicine (preferably with exposition and immunotherapy or hyposensitization). Anxiety therapies would improve qualitatively and could be cut down in duration.


[A comparison of East German and West German probands: emotionality and objective personality variables].

Data from a joint research project between the University of Trier and the Humboldt University Berlin of personality dimensions were compared and analyzed. Since Germany was separated for forty years, studies of the general population are necessary in order to yield possible cultural differences. In the present comparison we analyzed personality constructs like anxiety, curiosity, anger and anger expression as well as variables of objective personality tests. The sample from the former western German states contained N = 109 persons. As a basis for comparison, we selected a parallized matched sample (n = 109) from the new eastern German states out of a larger sample (N = 586). The main results were as follows: (1) reliability (consistency) and factor structure of the measures are similar for both samples. This equivalency is the formal prerequisite for a comparison. (2) There are substantial differential effects between eastern and western samples. Wellness as well as emotionality also show different results considerable with regard to age and sex.


[Ethical aspects of psychotherapy].

There are many ethical problems in theory and practice of psychotherapy. Aims of therapy, effectiveness, side effects and modalities of therapy are discussed in detail. The dependency of these aspects from anthropological considerations and understanding of illness is stressed. As far as side effects of psychotherapy are concerned abuse of the patient by the psychotherapist is distinguished from those potentially harmful effects which are due to the specific process of psychotherapy. The relationship of psychotherapy to psychopharmacotherapy is discussed in a special passage.


[A multi-stage cognitive training program for improving cognitive functions of chronic alcoholic patients].

In accordance with neuropsychological findings, that chronic alcohol abuse can lead to cognitive impairment, a multi-phased cognitive training program (MKT) developed by Sagstetter was conducted with a group of chronic alcoholic patients. In this article this program is introduced and evaluated with regard to chronic alcoholic patients and their improvement in certain cognitive functions. The study group consisted of 11 chronic alcoholics, who regularly took part in the program. The control group consisted of 11 patients who took part on a behavior therapy program offered by the clinic and had similar social and pathological histories to those of the study group. The two groups were compared with each other with the help of a follow-up study design after approximately seven weeks. The results reveal that this present cognitive training program promotes, above all, cognitive functions such as visual short-time memory, concentration, form perception as well as combination and organisation ability.


[The hypochondriacal delusion].

Hypochondriacal delusions are not bound to a specific mental disorder, but occur in the whole spectrum of psychoses. In spite of the implicitly wrong conception of reality, the significance of these delusions as an attempt to express, interpret and cope with mental illness should not be neglected if an approach to the patient's own experience is sought. On the one hand, hypochondriacal delusions are based on altered body perceptions in mental illness, characterized by primary local or general dysaesthesias to the point of depersonalisation, or caused secondarily by the patient's increased attention to his own body. This alienated bodily experience is named in (lay) medical terms by the delusional hypochondriac. On the other hand, "illness" with its manifold cultural and psychological meanings stands as a polyvalent metaphor for the incomprehensible alteration of the patient's self-experience in general: thus, the feeling of exposure to external attacks, of guilt, sinfulness, shame, loss of self-esteem or even the experience of a disruption of personality may find their expression in the hypochondriacal delusion.


[German version of the Vanderbilt Psychotherapy Scales: description and use in two brief psychotherapies].

The Vanderbilt-Psychotherapy-Scales appeared to be useful instruments for the identification of process characteristics determining the outcome of brief dynamic psychotherapies. The German versions of the two scales measuring important components of the psychotherapeutic process and of negative indicators are described in this article. It could be shown that--similar to the original versions--both scales can be used economically and that rater agreement soon reaches satisfactory levels. To demonstrate the potential use of the scales, their application in two different short term treatments are described which are presently analysed in a multi-center study of "single case oriented process research" using a variety of different methods. The example demonstrates that both scales are useful to discriminate both different theoretical foundations of the treatments as well as specific features of the patients and their therapists. The usefulness of the scales in a broader context is mainly based upon the experience that they reveal a lot of important informations about the psychotherapeutic process with a minimum of expense.


[In situ detection of EGF receptor mRNA in arteriosclerotic lesions in man: implications for the proliferative activity of smooth muscle cells].

Growth factors and growth factor receptors are considered to be key elements in the pathogenesis of arteriosclerosis and restenosis formation. To study the local expression of epidermal growth factor (EGF) receptor, plaque tissue specimens from advanced lesions (10 coronary, two femoral, seven carotid) of 19 patients were taken for in situ hybridization studies using an EGF-specific cDNA probe. In serial vascular sections of three lesions with increased focal cellularity, autoradiographic silver grains were clearly localized to intimal cells adjacent to the internal elastic lamina. EGF mRNA transcripts were not observed in the fibrous cap, the plaque shoulders, necrotic intimal areas, or in the media. In smooth muscle cells (SMCs) cultured from human plaque tissue, EGF increased SMC proliferative activity in a dose-dependent manner (ED50: 3-6 ng of EGF/ml). Proliferative responsiveness to EGF (10 ng/ml) was found to be significantly (p < 0.01) enhanced in coronary SMCs derived from restenotic lesions as compared to those from primary stenoses. The expression of EGF receptor mRNA in human atheromatous lesions could be of prognostic value to predict an increased SMC proliferative response to stimulatory growth factors.


[The anti-ischemic effect of gallopamil-retard in comparison with nifedipine-retard in stable angina pectoris].

To assess the antiischemic efficacy of slow-release (SR) gallopamil, 100 mg b.i.d., versus slow-release (SR) nifedipine, 20 mg b.i.d., 24 patients with chronic stable angina underwent symptom-limited bicycle ergometer exercise stress tests in a randomized, placebo-controlled, double-blind, cross-over protocol. Both medications caused a significant reduction in anginal attack frequency and nitroglycerin consumption as compared to placebo; similarly, exercise tolerance was augmented in association with a considerable reduction in ischemia-induced ST-segment depression. The antiischemic effect of gallopamil (SR) was marginally superior to that of nifedipine (SR). Since the incidence of adverse effects was also less with gallopamil (SR) this drug exhibited a more favorable risk-benefit ratio relative to nifedipine (SR).


[Interval resistance exercise in comparison with bicycle ergometry stress. Studies with resistance endurance training in coronary patients].

In the rehabilitation of coronary patients there is an increased interest in using complementary resistance exercise training. Therefore, we studied nine patients (males; age: 51 +/- 7 years) with chronic stable coronary heart disease during extensive resistance exercise (ex RE) (legpress, abduction, adduction) (60-s work: 60-s rest; contraction intensity: 65% of 1 RM) and during intensive resistance exercise (int. RE) (legpress) (30-s work: 45-s rest) with 85% of 1 RM. Non-invasive continuously measured blood pressure, heart rate, norepinephrine, epinephrine, lactic acid, and glucose were compared with values from maximal bicycle ergometry (3-min steps, each 25 w; max. performance: mean 156 w; range 125-200 w). 1) Comparing ex RE and int RE with bicycle ergometry there were no differences in blood pressure (systolic: 206 and 204 vs. 210 mm Hg; ns; diastolic: 98 and 104 vs. 92 mm Hg; ns). Heart rates (104 and 103 vs. 125/min; p < .01), norepinephrine (3.8 and 3.3 vs. 8.8 nmol/l; p < .01) and epinephrine (0.7 and 0.6 vs. 1.4 nmol/l; p < .01) were considerably lower. 2) The most significant increase and decrease of blood pressure and heart rate occurred within 15-30 s after the beginning and end, respectively, of isometric exercise. 1) ex RE is suitable for patients with stable CHD and cardiac exercise tolerances of 1.5-2 W/kg = 125-150 watts. 2) Blood pressure monitoring by the cuff method (RR) immediately after RE did not reflect blood pressure during RE. 3) Controlling RE by the training heart rate prescribed for endurance exercise is not possible.


[Rotational stability of angiography catheters].

Rotatory stability is a parameter that reflects the ability of a catheter to transmit a rotation applied at the outer end to the catheter tip for the purpose of selective probing. A method for measuring the rotatory stability is described, and the results of rotatory stability measurements of 70 different commercially available catheters are reported. There is an almost linear correlation between the rotatory stability and the difference between the respective fourth power of the external and internal diameter or, approximately, to the fourth power of the external diameter for catheters without wire reinforcement. With the same cross-sectional dimensions, the rotatory stability of teflon, polyethylene, and nylon catheters has an approximate ratio of 1:2:4. Wire reinforcement increases rotatory stability by an average factor of about 3. For catheters of calibers 5 F and 6 F, a correlation between the rotatory stability and the weight of the reinforcing wire mesh is apparent.


[Peripheral embolism of hemostasis collagen (VasoSeal)].

VasoSeal is a purified bovine, absorbable collagen plug currently successfully used to close the femoral arterial puncture site after cardiac catheterization under full anticoagulation. Up to now there has been no experience with potential complications. We observed acute ischemia in the right lower leg of 2/100 patients 36 resp. 24 h after successful closure of the puncture site with VasoSeal. Angiography confirmed acute occlusion of the distal A. poplitea dextra. A 25-mm resp. 50-mm long cylindrical foreign body embolus was removed with a Fogarty-catheter by retrograde indirect embolectomy. Histopathology confirmed a fresh collagen clot with appositional thrombosis.


[Pimobendan (UDCG 115 BS) in long-term therapy of chronic heart failure].

Pimobendan is a positive inotropic agent with additional calcium-sensitizing effects of the phosphodiesterase III-inhibitor group. In short-term studies, beneficial hemodynamic effects have been demonstrated in patients with congestive heart failure. The aim of this prospective study was to examine the long-term effect of pimobendan (during at least 6 months) on subjective state, hemodynamic parameters, and arrhythmias in patients with congestive heart failure NYHA classes II and III. After double-blind randomization, 24 patients received pimobendan 5 mg bid or placebo orally in addition to a basic therapy (diuretics, digitalis). After 3 months, pimobendan-treated patients showed a significant clinical improvement (p < 0.03). In the placebo group, one patient underwent acute cardiac transplantation due to rapid clinical deterioration; another patient died suddenly after 5 months. No cardiac events occurred in the pimobendan group. In comparison to placebo, no proarrhythmogenic effect of pimobendan was detected. Clinical stabilization of patients in the pimobendan group was not paralleled by improvement of the hemodynamic parameters of left-ventricular performance.


[Function of the right ventricle in patients with dilated cardiomyopathy].

To analyze right-ventricular size and function and their relationship to left-ventricular dimensions in patients with dilated cardiomyopathy (DCM), biplane cineventriculography was performed in 57 patients. The results were compared to 15 normals (N). In patients dilatation of the right ventricle (RVEDVI: DCM: 126.5 +/- 41.4 ml/m2, N: 90.5 +/- 9.2 ml/m2, 2 p < 0.05) was less pronounced than dilatation of the left ventricle (LVEDVI: DCM: 136.0 +/- 45.8 ml/m2, N: 76.7 +/- 7.9 ml/m2, 2 p < 0.05). Left-ventricular ejection fraction (LVEF: DCM: 36.1 +/- 10.2%, N: 64.4 +/- 3.8%, 2 p < 0.05) was more reduced than right-ventricular ejection fraction (RVEF: DCM: 39.7 +/- 11.5%, N: 58.3 +/- 3.3%, 2 p < 0.05). Concerning the individual patient, a good correlation was found between right- and left-ventricular stroke volume (r = 0.74), whereas ejection fraction (r = 0.58), enddiastolic (r = 0.52) and endsystolic volume (r = 0.55) of the left and right ventricle correlated only moderately. Twenty-three of the 57 patients showed pronounced differences between right- and left-ventricular ejection fraction. The difference RVEF-LVEF was < = -10% in six patients, i.e., right-ventricular ejection fraction was markedly more reduced than left-ventricular ejection fraction. Right-ventricular myocardial biopsy was performed in five of these six patients with histologic evidence of dilated cardiomyopathy and, also, no signs of right-ventricular dysplasia (no lipomatous tissue replacement).(ABSTRACT TRUNCATED AT 250 WORDS)


[Initial presentation of ventricular pre-excitation after catheter ablation of concealed retrograde conducting accessory atrioventricular pathways].

Catheter ablation using radiofrequency or direct current energy was performed in 19 consecutive patients with concealed accessory pathways. Four patients developed manifest preexcitation for the first time following ablation. The concealed accessory pathways of these four patients were localized at right lateral, left posteroseptal, left posterolateral, and left lateral sites, respectively. In two patients, manifest preexcitation developed immediately after attempted ablation. In the other two patients, preexcitation occurred after 4 and 14 days. Electrophysiologic testing revealed in all four patients that the origin of manifest preexcitation corresponded to the site of the concealed pathway. Successful catheter ablation using radiofrequency current was performed in three patients. One patient preferred surgical interruption of the accessory pathway. The reason for this sudden emergence of preexcitation of these patients with only retrograde conducting pathways remains to be investigated. The anterograde conduction capacity might have been modified by damaging the site of the accessory pathway insertion.


[Successful treatment of atrial fibrillation by resection of a congenital aneurysm of the left heart atrium].

Atrial fibrillation occurred in a 27-year-old patient with a history of globular cardiac enlargement since childhood. Because of the probable causal relationship between the preexisting heart disease-which was supposed to be an enlargement of the left atrium-and the rhythm disturbance, we recommended a surgical intervention. Cardiac surgery revealed a congenital aneurysm of the left atrial appendage which could be resected without any complication. Postoperatively, atrial fibrillation had returned to regular sinus rhythm. The bad prognosis with a high risk of systemic embolism is the reason why early cardiac surgery should be performed after diagnosis of this rare anomaly (20 reported cases) of the left atrium.


[Effectiveness of propafenone in congenital ectopic junctional tachycardia--a case report].

Junctional ectopic tachycardia is associated with a poor prognosis when it occurs in newborns and young infants. Like other automatic tachyarrhythmias, junctional ectopic tachycardia has been shown to be very resistant to medical treatment. Successful therapy with propafenone in a newborn with congenital junctional ectopic tachycardia is presented. Due to its high effectiveness, safety, and lack of side-effects, propafenone appears to be a valuable drug in the treatment of young patients with congenital junctional ectopic tachycardia.


[Percutaneous high frequency current catheter ablation in permanent ventricular tachycardia of the "bundle-branch reentry" type after implantation of an automatic cardioverter-defibrillator].

A 65-year-old female patient with a history of recurrent sustained ventricular tachycardia presented with an incessant ventricular tachycardia (cycle length 360-400 ms) following implantation of a cardioverter-defibrillator (ICD). The tachycardia could not be terminated by antiarrhythmic drug treatment, antitachycardia pacing or internal defibrillation via the ICD. An invasive electrophysiologic study revealed that the mechanism of this newly occurring tachycardia was bundle branch reentry. The patient underwent emergency catheter ablation using radiofrequency (RF) current. Endocardial mapping of the right bundle branch and of the distal His bundle was performed and a bundle branch reentry tachycardia was diagnosed. After delivery of the fifth RF-impulse, the tachycardia terminated and complete AV block was induced. No malfunction of the ICD was observed following RF-ablation. The patient was hemodynamically stable with a junctional escape rhythm and antibradycardia pacing back-up of the ICD (VVI-mode). This case report demonstrates the feasibility of RF catheter ablation in the treatment of incessant bundle branch reentry tachycardia as a complementary option after implantation of an ICD.


[Unexpected malposition of a ventricular pacemaker electrode].

Ten years after mitral-valve replacement, a 69-year-old patient underwent VVI pacemaker implantation because of symptomatic bradyarrhythmia. Postoperatively, the electrocardiogram showed a right bundle branch pattern under ventricular stimulation while the threshold was optimal. Under fluoroscopy, we suspected a malposition of the electrode outside the right ventricle in the LAO and lateral view. This could not be verified by echocardiography, whereas contrast angiography of the right ventricle proved the extracavitary position of the electrode under the inferior wall of the left ventricle, probably in the middle cardiac vein. The electrode position was operatively corrected without complications. We discuss different ways of malposition and their detection by considering electrocardiographic configuration and fluoroscopy in LAO and lateral view.


[Pulmonary valve atresia with ventricular septum defect: interventional recanalization of the right ventricular outflow tract].

A case of pulmonary atresia with ventricular septal defect is reported where continuity between the right ventricle and the hypoplastic pulmonary artery was established interventionally. The atretic valve was perforated using a special "perforation needle" with a sharp and stiff distal and a flexible proximal part. Perforation of the bifurcation was well tolerated without later sequelae. After perforation of the atresia, dilation was successfully performed using 2, 4, and 7.2 mm balloons with a pressure of 10 atm; the arterial oxygen saturation increased from 72% to 84%. Four weeks later repeated "valvuloplasty" was performed (balloon diameters 8 mm, 9.5 mm, and 12 mm) and the "valve" ring was dilated to a diameter of 10.5 mm. Although no general conclusions can be drawn from this single application, mechanical perforation of the atresia could become an attractive interventional approach for the treatment of pulmonary atresia.


[Acute and intermediate clinical and angiography results after coronary stent implantation].

The value of stenting in emergency situations in interventional angioplasty procedures is widely accepted. Stenting remains controversial as a therapeutic method for use beyond the acute phase, as the results reported in the acute phase are characterized by a wide range. This study reports the implantation results of Palmaz-Schatz stents in 64 cases mostly for failed PTCA. Stents were mounted on Monorail-PVC-balloon catheters. The acute success rate was 96.8%. In two cases with stent misplacement CABG was performed within 48 h. No lethal events occurred in the acute phase. Stent thrombosis has only been seen in four cases (6.5%) so far under subtle monitoring of the anticoagulant regime. Restenosis rate (definition: NHLBI-4) according to serial angiography 6 and 12 weeks after implantation was 4.25% at 6 weeks, with a cumulative rate of 19.4% up to 12 weeks. All recurrences eligible for PTCA have been successfully redilated. A high extracardiac complication rate--predominantly groin hematoma--is the main disadvantage of the procedure. Surgical repair was necessary in 11.4% up to 6 weeks after implantation. Surprisingly high was also the incidence of hematuria, also in 11.4%, in the phase of high anticoagulation. In conclusion, stenting is an effective therapeutic method to avoid emergency CABG and elective CABG in most cases of failed PTCA.


[Randomized comparison between collagen administration and pressure dressings for occlusion of the arterial puncture after coronary angiography and coronary dilatation].

One hundred patients undergoing routine diagnostic or interventional catheterization were randomly assigned to receive either percutaneously applied collagen (group A; n = 50) or conventional pressure dressing (group B; n = 50) for sealing of the femoral artery. Clinical variables were comparable in both groups. The heparin dose was 100 IU/kg in 30 patients and 200 IU/kg in 20 patients of both groups. The average compression time was 4.3 min in group A and 42.3 min in group B (p < 0.001). In group A, 1/50 patients required pressure dressing compared to 50/50 patients in group B. Bleeding was not observed in group A, but was observed in 6/50 patients in group B. The time to ambulation was 6.4 (range, 4-12) h in group A and 21.6 (range, 10-48) h in group B (p < 0.001). Hematomas with a diameter of > 6 cm developed in 4/50 patients in group A and in 11/50 patients in group B (p < 0.05). Blood-transfusions or surgical interventions were not required and there was no loss of ankle pulses in either group. In conclusion, percutaneously applied collagen reduced compression time and duration of bedrest after diagnostic catheterization and PTCA. Despite earlier ambulation, the incidence of bleeding was lower with collagen than with conventional pressure dressing.


[Spontaneous renal artery dissection].

The clinical clues of sudden arterial hypertension, acute pain in the side, proteinuria, hematuria, abdominal bruits and renal failure are suspicious for spontaneous dissection of the renal arteries, even in the absence of one or more of the above-mentioned symptoms. To confirm the diagnosis and to provide appropriate therapy, immediate renal arteriography is mandatory, otherwise acute loss of renal tissue may occur. We report on one patient with spontaneous dissection of the renal arteries.


[Geriatric diseases of the upper digestive tract].

During aging, secretion and motility of the upper GI tract slow down. The reduction of these functions, however, does not create complaints. In the higher age groups, a number of symptoms from age-dependent diseases occur more frequently, e.g., dysphagia in response to cerebral ischemia, or disturbed gastric emptying caused by diabetic visceral neuropathy. Moreover, certain GI diseases occur more often in the elderly, e.g., chronic atrophic gastritis, NSAR-induced gastric ulcers, malignancies, and others. In contrast, almost nothing is known about diseases or symptoms of the GI tract that might be specific for the elderly. With only a few exceptions, there are no age-dependent clinical differences. Nevertheless, intestinal diseases often develop more rapidly and the mortality is higher in the elderly than in younger people.


Cortical bone age estimates from historically known adults.

Adult skeletal remains from 19th century historic cemeteries were used to test the accuracy and precision of age estimations based on the cortical bone histological technique proposed by Thompson [6, 7]. Comparison of estimates made from anterior microscopic fields versus peripheral fields (N = 29 femora) indicates general age agreement, with 6/29 demonstrating marked differences between sampling locations. Three cases of unilateral trauma show no consistent effect on bone age estimates. Among sixteen adults of known age, the ages of eight were accurately estimated; five were estimated to be significantly older and three were estimated to be younger than they were. It is argued that cortical erosion/diagenetic destruction may contribute to overestimation by obscuring the periosteal regions required for application of the technique. While not without its shortcomings, it is concluded that the Thompson technique is relatively robust and merits further development.


[Gastrointestinal motility in the elderly].

The existence of specific, age-related changes in gastrointestinal motility with clinical significance is controversial. Beside the more infrequent primary motility disorders, secondary motility disturbances associated with collagen vascular diseases, endocrinopathies, and neuromuscular diseases are prominent in the older and often multimorbid patients. Especially in geriatric patients, motility associated symptoms are undesired side-effects of drug therapy. The pathophysiology, clinical syndromes, and therapeutic principles of motility disorders in the elderly are discussed. The major symptoms of esophageal dysfunction are dysphagia, chest pain, heartburn, and regurgitation. Oropharyngeal dysphagia, mostly caused by cerebrovascular accidents and other neurologic disorders, leads to disturbances in food intake, and is often complicated by broncho-pulmonary infections arising from recurrent aspiration of food or saliva. Gastrointestinal reflux disease and spastic motility disorders of the esophagus are regarded as possible causes of angina-like chest pain after exclusion of cardiac diseases. Motility disturbances of the stomach and small bowel are often related to systemic disease (i.e., diabetes mellitus, chronic intestinal pseudo-obstruction) of drug side-effects. Mental and physical decline, reduced fluid intake, and constipating drugs are the most relevant factors for idiopathic constipation in the elderly. Fecal incontinence means a great psychological strain for older patients and leads to social isolation.


Incisification of anthropoid deciduous mandibular canines.

Interspecific variation of the deciduous mandibular canine's mesial cristid in 51 anthropoid taxa is assessed. In 84% of the taxa observed, mesial cristids were well developed and provided incisor-like occlusion with di2. In 5 of the remaining 8 taxa, lingual cristids provided incisor-like occlusion with dc1. Comparisons between dc1s and C1s of conspecific adult males and females show that in 85% of the species observed the magnitude of mandibular canine incisor traits follows this order: deciduous > adult female > adult male. This within-species variation is inversely related to the likely adaptive value of the canine as a weapon and to canine crown height. A selection model based on these data is derived and applied to human canine evolution. The relevance of these data to the field vs. clone theory debate about dental development is also discussed.


[Calculi of the bile ducts--a problem for the elderly? Observations from the surgical viewpoint].

Age alone does not increase the risk in biliary surgery. However, cholelithiasis in the elderly is often associated with an increased frequency of acute cholecystitis and cholangitis due to stone obstruction. The proportion of elderly people in the world is still growing, therefore, we can expect to see increasingly more complications of gallstones. Elective surgery for gallstone disease in all age groups is to be preferred to a policy of waiting for stone complications with the resultant risk of higher mortality and postoperative morbidity of emergency surgery.


Estimation of stature in children from second metacarpal measurements.

On the basis of 552 boys and 542 girls aged 6 to 20 years, this study examines the estimation of stature from dimensions and maturity of second metacarpals by means of linear regression equations. A combination of length and width measurements provided a more accurate estimation than each measurement individually. When taken alone, length produced a more accurate estimation than width. Sex and age factors are useful for the estimation of stature, though these variables are often unknown in the isolated bone. The samples are divided into immature and mature groups (according to skeletal maturity). Regardless of sex, stature could be estimated from the metacarpal length and width with a standard error of 4.19 cm by means of a multiple linear equation in the immature group. The mature group should be considered with adults for this purpose. Thus, taking into account their skeletal maturity, living stature could be practically estimated from the second metacarpal with significant degrees of accuracy in children.


[The small and large intestine. Fecal incontinence].

Except for impaired resorption of vitamin D and calcium, the small bowel does not exhibit significant functional impairment during aging. Vascular problems in the small and large bowel are rare, but are of dramatic importance. Chronic constipation and diverticulosis are classical disorders of aging which should be treated by adding fiber to the normal diet. Surgery is only indicated in the case of complicated diverticulitis. Adenoma and colorectal carcinoma show an age-related incidence. Angiodysplasia, preferentially localized in the caecum, is the major cause of lower gastrointestinal tract hemorrhage. Anal incontinence may be multifactorial, behavior modification and biofeedback may help to avoid surgery.


[Housing for the elderly--results of an empirical study].

This empirical study is based on 285 standardized interviews with older people living in government-subsidized housing (sozialer Wohnungsbau). Housing conditions, especially contentment and perceived deficiencies are described. Additional remarks refer to the neighborhood relations. The high contentment, already known from former studies, is confirmed by the results. The findings indicate that steps towards an adaptation of housing conditions to possible needs of older people are not ranked very high. Beyond that, there seems to be a lack of information about the technical possibilities of improving the housing conditions, although older people want to live as long as possible in their current housing.


"Lucy's" body height and relative leg length: human- or ape-like?

The body height of Australopithecus afarensis A.L. 288-1 ("Lucy") has recently been estimated and calculated as between 1 m to 1.06 m; other estimates give ca. 1.20 m. In addition, it is often stated that her relative leg length was shorter than that of modern humans. Using relative leg-, femur- and tibia length it is shown that both statements together can not be true; either her body height must at least have been around 1.06 to 1.10 m to give "Lucy" human-like leg proportions, or, to achieve a shorter, more ape-like leg ratio, a body height of ca. 1.20 m must be assumed.


Changes in thyroid hormones and serum lipids of aged women following thyroidectomy.

Thyrotropin (TSH), thyroxine (T24) 3,5,3403 triiodothyronine (T23), serum lipid levels and ECG findings in females who underwent thyroidectomy 2-44 years earlier, were compared to those of similar controls with intact thyroid. Pathological ECG is more frequent in thyroidectomized persons who also exhibit higher thyrotropin and lower HDL-cholesterol levels. The results indicate that thyroidectomized state should be regarded and controlled as a possible condition of subclinical hypothyreosis, being a risk factor of cardiovascular diseases.


Influence of age and sex on 19 blood variables in healthy subjects.

Normal values exist for all clinical chemical tests, but it is not very clear what is normal for healthy elderly subjects. Therefore, routine blood variables were determined in 80 ambulatory, disease-free persons who had undergone rigorous health screening. The subjects were divided into the following age groups: 20 (+/- 3), 40 (+/- 3), 60 (+/- 3), and 80 (+/- 5) years, with 10 males and 10 females per age group. Blood variables were determined after an overnight fast. It was found that even with conservative statistical measures more than half of the variables were significantly affected by age or sex. Significant age differences were found for total cholesterol, triglycerides, sodium, and ASAT. Urea, creatinine, gamma-GT, phosphate, alkaline phosphatase, and albumin were characterized by both age and sex differences. No age or sex differences were found for glucose, potassium, chloride, calcium, calcium ion, iron, magnesium, total protein, and ALAT. The findings suggest that the age or sex-related changes of a number of blood variables such as cholesterol, triglycerides, and liver enzymes are not only of statistical significance, but are also of clinical relevance.


[The significance of current hepatitis virology for geriatrics].

In geriatric medicine, hepatitis A is of minor interest only because protecting antibodies have been demonstrated in 92% of the elderly. Hepatitis B (which, prior to recognition of major routes of transmission, was predominantly transmitted percutaneously) has gained importance only by its chronic residuals in aged patients; acute infection in the elderly is rare. The recently detectable posttransfusional hepatitis C is the most important type of viral hepatitis in the elderly. In our region, hepatitis D is rare and hepatitis E has not yet been observed.


[Thyroid gland disorders in elderly psychiatric patients].

With 200 admissions of 192 patients to the psychogeriatric acute ward of the Zwiefalten State Mental Hospital serial measurements of serum concentrations of basal TSH and, if necessary, of total T3 and total T4 were carried out with the TRH tests performed. Basal TSH was decreased in 16% of patients and in 2% it was increased. Only three patients had manifest hyperthyroidism and one patient had marginal hypothyroidism. Regardless of their psychiatric diagnosis, patients with manifest hyperthyroidism showed psychomotor agitation, whereas those with latent and marginal hypothyroidism suffered from lack of impulse. These symptoms improved after treatment with carbimazol or levothyroxine, respectively, and psychotropic drugs. Therefore a thyroid screening should be performed on psychogeriatric patients regardless, including, when possible, repeat measurement of basal TSH and, if necessary, the test battery should include total T3 and total T4.


Secular trend of menarcheal age in southern Chinese girls.

Southern Chinese girls aged 11 years and 9 months to 12 years and 3 months in Hong Kong have a mean menarcheal age of 11.50 years (standard deviation of 0.47) using the recollection method. Highly significant differences are found when compared to the 12-year-old girls in Hong Kong studied in the past decades. Therefore, a secular trend of earlier menarcheal age is demonstrated.


Emergence of deciduous teeth in Punjabi children, north India.

Data on deciduous tooth emergence of 312 children aged 4 to 31 months of Punjabi parentage are presented. Probit analysis was used to derive the median age of tooth emergence. Female children are found to be advanced with respect to tooth emergence than their male counterparts. While comparing the present data with those from other populations it is found that, in general, the mean number of emerged teeth in Punjabi children is more at most ages, with lower median age of eruption for most teeth. Magnitude of interage variability in the eruption times is noticed to be maximum in the 16-17 and 20-21 months age groups. The findings of the study suggest that number of teeth can be used as a parameter for the estimation of age.


Asymmetry and sexual dimorphism of lungs weight in fetal ontogeny.

1079 male fetuses and 727 female fetuses at the age of 20 to 41 weeks were investigated for the process of asymmetry and sexual dimorphism of lungs weight formation as well as developmental correlation between the weight of the lungs and the size of the heart. Statistical analysis of the results was applied. It was ascertained, among others, that asymmetry of lungs weight occurs in the investigated developmental period--the right lung is heavier than the left one about 20 to 30%. Between the increase in the weight of the lungs and the size of the heart positive correlation occurs, but it is not of directed character. A substantial, intersexual differentiation of lungs weight was not ascertained.


Origin of the Romany gypsies--genetic evidence.

Genetic heterogeneity and affinity was examined between nine Romany speaking gypsy populations and two of their possible ancestral populations from India. For AB0, Rhesus (D), MN and HP systems there exists a conclusive heterogeneity among these populations. Three gypsy populations from Western Europe (English, Welsh and Swedish) are genetically distinct from the rest of the East European gypsies and the populations of India analysed in this investigation. Overall the genetic differentiation among these populations is moderately high (RST = 0.029). The results also indicate the relative close relationship among the East gypsies and the two selected nomadic populations of India. The factors responsible for the moderate diversification of the East European gypsies may be high rate of migration, isolation and random drift, while among the Western gypsy populations admixture seems to be an important differentiation factor.


Flatness of facial skeletons in Siberian and other circum-Pacific populations.

Thirty-four populations from Siberian and other circum-Pacific regions were compared in terms of facial flatness measurements of the cranium. While fundamentally having an extremely flat face, Siberian populations tend to be differentiated into two or three subgroups. On the other hand, other Mongoloid populations show greater variation in facial flatness. The less flat faces of the American Indians are almost equal to those of the Europeans. In Japan, the existence of two contrasting groups in terms of facial flatness have been found.


Investigations on the variability of blood group polymorphisms among sixteen tribal populations from Orissa, Madhya Pradesh and Maharashtra, India.

Sixteen tribal populations from Orissa, Madhya Pradesh and Maharashtra have been typed for the polymorphic blood group systems A1A2B0, MNSs, Rhesus, Kell, Duffy and Diego. The heterogeneity in the distribution of haplotype and allele frequencies, respectively, is partly considerable. It is supposed that this is due to the operation of several microevolutionary factors, such as genetic drift, social and geographic isolation and gene flow. This is discussed in detail.


Apes and apomorphies: the anterior nasal spine as a projection of cladistic conceptions.

Projections, tubercles, or spines of bone at the anterior margin of the piriform aperture occur in widely varying forms among catarrhine primates. This paper describes, illustrates, and tabulates the frequencies of such structures by age and sex in a large collection of Pan troglodytes verus skulls. In the overall sample of 263 skulls where the region could be observed, these protuberances of bone occur bilaterally in 10.7% of specimens and unilaterally in an additional 3.4%. They increase in frequency with age to a high of 16.9% in adult and older animals. Among adult specimens, these projections of bone in the anterior nasal region are present more commonly in females than in males. In a comparative context, variants on this structure in nonhuman primates, some of them documented previously, should help to counter the cladistic conception that the anterior nasal spine is an apomorphic or evolutionary novel morphological structure in hominids.


Experimental evaluation of antitoxic protective effect of new cholera vaccines in mice.

Intraperitoneal immunization of mice and subsequent challenge with purified cholera toxin (CT) were employed to evaluate the anti-cholera toxin protective effect of two new oral cholera vaccines, live CVD 103-HgR and killed B subunit-whole cell (BS-WC). CVD 103-HgR vaccine demonstrated 100% protection of mice against 2.25 LD50 and 70% against 3 LD50 of CT. Mice immunized with BS-WC vaccine were protected against 2.25 and 3 LD50 of CT in 88 and 62% of cases, respectively. All three killed parenteral vaccines failed to protect against CT. We suggest this mouse system for preliminary evaluation of the antitoxic protective activity of cholera vaccines.


A single vaccination with an inactivated hepatitis A liposome vaccine induces protective antibodies after only two weeks.

In the near future an inactivated hepatitis A vaccine will be commercially available. The recommended vaccination schedule will include at least two vaccinations 1 month apart. Giving two doses of vaccine on the same day in one injection results in protection after 2-4 weeks dependent on the adjuvant used. Hepatitis A virus incorporated into liposomes proved to be a suitable formulation in term of rapid seroconversion, high level of mean antibody content and low reactogenicity.


Compulsory and non-compulsory immunizations: contraindications perceived by medical practitioners.

A total of 284 general practitioners (GPs), paediatricians and doctors of public vaccination centres (DPVC) were interviewed to investigate their willingness to immunize children with compulsory (diphtheria-tetanus, oral polio) and non-compulsory (measles, pertussis) vaccines in the presence of 19 different medical histories. We observed a reluctance to immunize in the case of false contraindications, a lack of information mainly about non-compulsory immunizations, and doubts about the real contraindications to polio, measles and pertussis vaccines. The frequency of correct answers to the question posed was significantly higher in the group with less than 20 years of experience, and the DPVCs proved better informed about immunization. However, the crucial role played by the GPs and paediatricians' advice can prejudice the correct use of active immunization.


Liposomes enhance the immunogenicity of reconstituted influenza virus A/PR/8 envelopes and the formation of protective antibody by influenza virus A/Sichuan/87 (H3N2) surface antigen.

Reconstituted influenza virus (A/PR/8 strain) envelopes (RIVE) and influenza virus (A/Sichuan/87 (H3N2) strain) surface antigens were entrapped in dehydration-rehydration vesicles (DRV liposomes) composed of egg phosphatidylcholine (PC) or distearoyl phosphatidylcholine (DSPC DRV) and equimolar (32 mumol) cholesterol. Entrapment values for RIVE were 31.2 (PC) and 29.4% (DSPC DRV) of the material used. Corresponding entrapment values for the A/Sichuan/87 strain antigens were 40.7 and 39.3%. Balb/c mice injected intramuscularly with PC or DSPC DRV liposomes containing 0.1 and 1.0 microgram RIVE exhibited primary (higher dose only) and secondary responses (IgG1) which were significantly higher than those obtained in mice injected with identical amounts of non-entrapped RIVE. Significantly higher secondary responses were also observed for the IgG2a and IgG2b subclasses. In experiments designed to assess the effectiveness of DRV liposomes as a carrier of influenza virus antigens in a potential vaccine, hamsters were immunized intramuscularly with 0.1, 0.5 and 5.0 micrograms of free or liposome-entrapped influenza A/Sichuan/87 surface antigens. Results showed increased haemagglutination inhibition (HI) antibody levels in terms of both primary (0.5 and 5.0 micrograms doses) and secondary (all doses) responses in the sera of animals treated with the liposomal formulations. DSPC compared with PC DRV exhibited greater adjuvanticity when the lower doses of antigens were used.


Inhibition of immune responses against rabies virus by monoclonal antibodies directed against rabies virus antigens.

Treatment of mice with a cocktail of murine anti-rabies monoclonal antibodies (mAb-C) interfered with the ability of these animals to mount a virus-neutralizing antibody response to rabies vaccine. Administered mAb-C did not affect the induction of rabies virus-specific T-helper cells. The magnitude of the inhibition of rabies virus-specific B-cell response was dependent on the concentration of the mAb-C and the duration of the mAb-mediated interference was inversely proportional to the biological half-life of the mAb. As long as the serum titres were above a critical threshold, the suppression could not be overcome even by multiple vaccinations. Since injection of mice with immunocomplexes consisting of inactivated rabies virus and mAb rendered the animals non-responsive to a subsequent vaccination with inactivated rabies virus, it is concluded that the mAb-induced suppression might be caused by the formation of antigen-antibody complexes which exert a negative signalling effect to premature B cells.


Genotypic and phenotypic characterization of an aroD deletion-attenuated Escherichia coli K12-Shigella flexneri hybrid vaccine expressing S. flexneri 2a somatic antigen.

The construction and characterization of EcSf2a-2, an aroD-deleted Escherichia coli-Shigella hybrid vaccine carrying chromosomal and plasmid genes from Shigella flexneri and expressing S. flexneri 2a somatic antigen in association with E. coli K12 core are described. Expression of hybrid lipopolysaccharide and deletion of aroD resulted in the attenuation of phenotypic characteristics associated with pathogenicity. The addition of an aroD deletion results in a requirement for an aromatic precursor of para-aminobenzoic acid (PABA), an essential bacterial metabolite not present in mammalian tissues. The biosynthesis of hybrid somatic antigen prevents expression of a Sereny-positive reaction by invasive bacteria capable of expressing a plaque-positive phenotype. A functional kcpA gene is required for expression of the plaque-positive phenotype. The presence of an aroD deletion does not interfere with expression of an invasive phenotype; however, in bacteria containing a functional kcpA gene, replication and spread by invading bacteria are limited, preventing development of the plaque-positive phenotype.


B30-MDP, a synthetic muramyl dipeptide derivative for tumour vaccination to enhance antitumour immunity and antimetastatic effect in mice.

The effect of a muramyl dipeptide derivative (B30-MDP) on the augmentation of antitumour immunity against highly metastatic L5178Y-ML25 mouse lymphoma cells was examined in CDF1 (Balb/c x DBA/2) mice. Mice immunized with a mixture of X-irradiated tumour cells (10(3)) and B30-MDP (100 micrograms) on 7 days prior to challenge by viable tumour cells displayed a significant decrease in metastasis towards the target organs, liver and spleen, compared with that of untreated mice. Immunization of mice with the mixture on day 5 or 7 after tumour challenge, when the level of glutamic-pyruvic transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT) in sera of mice inoculated with viable tumour cells was observed to be normal, caused less metastasis than immunization with X-irradiated tumour cells alone. Sensitization with X-irradiated tumour cells admixed with B30-MDP induced almost two times higher cytotoxicity of spleen cells against L5178Y-ML25 lymphoma cells than sensitization with X-irradiated tumour cells without B30-MDP. In contrast, cytotoxic activity of spleen cells against another target, L1210 lymphoma cells derived from BDF1 mice, was not observed by immunization with X-irradiated L5178Y-ML25 cells with or without B30-MDP. Specific lysis by splenic cells of the immunized mice against L5178Y-ML25 cells decreased to the normal level when T cells were deleted from the immunized spleen cells by the treatment of rabbit anti-mouse Thy1.2 antibody and rabbit complement. These results indicate that B30-MDP is able to augment a specific tumour immunity due to the enhancement of cytotoxicity mediated by T lymphocytes, and is useful as an immunopotentiating agent for active immunization of inactivated tumour cells.


Immune response to hepatitis B vaccine in parenteral drug abusers.

Responsiveness was assessed to a programme of vaccination of hepatitis B vaccine in a cohort of 197 intravenous drug addicts (mean age, 23.7 years) and their antibody response was compared with that of 271 healthy controls (mean age, 24.2 years). All participants were seronegative for hepatitis B surface antigen (HBsAg) and antibody to HBsAg (anti-HBs). The vaccination schedule consisted of three intramuscular injections (deltoid area) at months 0, 1 and 2. Although 70% of parenteral drug abusers received the three doses of vaccination, only 43.6% were evaluable for immune response. Fifty-eight per cent of heroin addicts and 80% of controls had evidence of anti-HBs seroconversion at 1 month after vaccination (chi 2 = 15.52, p less than 0.001). Geometric mean antibody titres were also significantly higher in controls (69.1 IU l-1; confidence interval 95%, 56.83 and 84.04) than in parenteral drug abusers (18.2 IU l-1; confidence interval 95%, 12.85 and 25.73) (F = 20.951, p less than 0.0001). The anti-HBs response was not influenced by coexistent anti-HBc, HCV antibody or HIV antibody seropositivity.


Effects of sample processing on the measurement of specific intestinal IgA immune responses.

The effects of techniques commonly used in the collection and processing of human intestinal fluid on the specific secretory immunoglobulin A (sIgA) response following oral immunization with the live typhoid vaccine Salmonella typhi Ty21a were examined. It was observed that the failure to adjust specific intestinal anti-typhoid lipopolysaccharide IgA antibody titres for total secretory IgA resulted in a false-negative detection rate of 19.8% and a false-positive detection rate of 7.4%. Furthermore, these specific responses were significantly diminished if the intestinal fluid was subjected to heat inactivation to reduce intestinal protease activity (p = 0.0083), but were not affected if stored at -70 degrees C for up to 1 year, without heat inactivation. It was concluded that in the processing of the intestinal fluid samples for specific sIgA determination heat inactivation significantly reduced specific sIgA titres, and that the failure to adjust absolute titres for total sIgA content resulted in a significant false-negative detection rate.


In vitro translation of a subgenomic mRNA from purified virions of the Spanish field isolate AST/89 of rabbit hemorrhagic disease virus (RHDV).

Purified preparations of the Spanish field isolate of rabbit hemorrhagic disease virus AST/89 were found to contain the plus-stranded genomic RNA of more than 7.4 kilobases (kb) and large amounts of a subgenomic mRNA of 2.4 kb. The smaller RNA was translated in vitro and shown to code for a 60 kDa protein which was immunoprecipitated using anti-RHDV as well as anti-VP60 sera.


A carboxy-terminal mutant spleen focus-forming virus (SFFV) envelope glycoprotein is transport-competent, but non-leukemogenic.

The Friend spleen focus-forming virus (F-SFFV) codes for a transport-defective leukemogenic envelope glycoprotein designated as gp52. We have previously shown that the external domain of gp52 carries the determinants responsible for its transport defect. Consistent with this idea, truncated gp52 molecules that lack a hydrophobic membrane anchor were transport-defective, and were not secreted from cells. In this report, we describe the construction of a mutant SFFV envelope gene that codes for altered gp52 molecules in which the carboxyl-terminal hydrophobic residues are replaced with exogenous hydrophilic residues encoded by the vector-derived sequences. The mutant env gene was expressed using the retroviral expression vector, pLXSN, and the mutant envelope protein was found to be transport-competent, and efficiently secreted from the cells. However, M-MuLV pseudotypes of the retroviral vectors expressing the mutant genome were found to be non-leukemogenic in mice.


Alterations in the levels of expression of specific cellular genes in adenovirus-infected and -transformed cells.

There is increasing evidence that changes in the transcriptional program of cellular genes in virus-transformed cells can contribute to virus transformation. It is, therefore, important to study altered expression patterns of cellular genes in adenovirus-infected and -transformed cells. We have used 40 different cellular genes or gene segments as hybridization probes to analyze the cytoplasmic RNA from adenovirus type 2 (Ad2)-infected KB cells, from Ad5-transformed human cells (293) or from several Ad2- or adenovirus type 12 (Ad12)-transformed hamster cell lines. Many of the genes probed were not expressed in human or hamster cells. Transcription of the ADPRT and the heat shock protein 70 genes was increased in Ad2-infected KB cells and in 293 cells. In Ad2-infected KB cells, c-myc gene transcription was decreased. In 293 cells and in three adenovirus-transformed hamster cell lines (T637, BHK21-Ad2E1A-E1B, and BHK21-Ad2 HindIII-G), the transcription of the c-jun gene was increased, whereas c-myc transcription was decreased in the latter two cell lines. The data presented here demonstrate that, among 40 different mammalian gene probes, alterations in steady state levels of RNA were detected for five of these genes. These results suggest major alterations in transcription patterns in adenovirus-infected and -transformed cells.


Mercaptoethanol and dithiothreitol decrease the difference of electrochemical proton potentials across the yeast plasma and vacuolar membranes and activate their H(+)-ATPases.

Mercaptoethanol and dithiothreitol (DTT) inhibited the acidification of external medium by Saccharomyces carlsbergensis cells and protoplasts during glucose oxidation. The inhibition was also observed when cells were incubated with mercaptoethanol or when mercaptoethanol and DTT were used to prepare protoplasts. Experiments with S. carlsbergensis plasma membrane vesicles and vacuoles showed these thiol reagents to inhibit ATP-dependent generation of delta pH and Em across plasma membrane vesicles and vacuoles but to activate their H(+)-ATPases. Mercaptoethanol and DTT are suggested to de-energize plasmalemma as well as tonoplast by increasing their H(+)-permeability and to disturb the cell ion homeostasis.


Genetic homology between Saccharomyces cerevisiae and its sibling species S. paradoxus and S. bayanus: electrophoretic karyotypes.

Chromosomal DNAs of many monosporic strains of the biological species Saccharomyces cerevisiae, S. paradoxus and S. bayanus were analysed using contour-clamped homogeneous electric field electrophoresis. Southern blot hybridization with eight cloned S. cerevisiae genes (ADC1, CUP1, GAL4, LEU2, rDNA, SUC2, TRP1 and URA3) assigned to different chromosomes was used to study homology and chromosomal location of the genes in the three sibling species. A comparative study of Ty1, Ty2 and telomere-associated Y' sequences having multiple chromosomal location was also done. Chromosome length polymorphism was found in cultured strains of S. cerevisiae. Wild S. cerevisiae and S. paradoxus strains yielded chromosome banding patterns very similar to each other. The karyotype pattern of S. bayanus was readily distinguishable from that of S. cerevisiae and S. paradoxus. Southern blot analysis revealed a low degree of homology between the S. cerevisiae genes studied and the corresponding S. paradoxus and S. bayanus genes. The number of chromosomes appears to be 16 in all three species.


Development of the yeast Pichia pastoris as a model organism for a genetic and molecular analysis of peroxisome assembly.

We describe the isolation of mutants of the yeast Pichia pastoris that are deficient in peroxisome assembly (pas). These mutants of P. pastoris can be identified solely by their inability to grow on methanol and oleic acid, the utilization of which requires peroxisomal enzymes, and are defined by the absence of normal peroxisomes as judged by electron microscopy and biochemical fractionation experiments. These mutants are the result of genetic defects at single loci and represent at least eight different complementation groups. The isolation of pas mutants of P. pastoris by a simple screen for mutants unable to use methanol and oleic acid represents a significantly more efficient method for identification of pas mutants than is possible in other organisms. To exploit this advantage fully we also developed new reagents for the genetic and molecular manipulation of P. pastoris. These include a set of auxotrophic strains with an essentially wild-type genetic background, plasmids that act as Escherichia coli-P. pastoris shuttle vectors, and genomic DNA libraries for isolation of P. pastoris genes by functional complementation of mutants or by nucleic acid hybridization. The availability of numerous pas mutants and the reagents necessary for their molecular analysis should lead to the isolation and characterization of genes involved in peroxisome assembly.


A screening procedure for the intracellular expression of native proteins by Saccharomyces cerevisiae: discrimination of diphtheria toxin-resistant mutants.

A general method is described for screening Saccharomyces cerevisiae colonies for the intracellular expression of native proteins. Colonies are replicated onto nitrocellulose membranes and yeast cell walls are removed enzymatically. The resulting spheroplasts are rapidly lysed by placing chromatography paper soaked in hypotonic buffer on the membranes. Intracellular proteins released by spheroplast lysis are bound in situ to the nitrocellulose under non-denaturing conditions and potentially can be examined using enzymatic or immunologic methods. For example, in the present study colonies were screened for the presence of elongation factor 2 (EF-2) that can be [32P]ADP-ribosylated by diphtheria toxin and [32P]NAD+. Recognition by the toxin requires the presence in EF-2 of the unique post-translationally modified histidine derivative, diphthamide. The procedure described here reliably discriminates between wild-type yeast colonies and mutant colonies that do not synthesize diphthamide. In addition to facilitating the study of diphthamide biosynthesis in yeast, the more general application of this procedure will enable the screening of colonies with assays that require native proteins.


Yeast flocculation: receptor definition by mnn mutants and concanavalin A.

Yeast flocculation involves the binding of surface lectins on flocculent yeasts, to carbohydrate receptors present as constituents of yeast cell walls. Receptors were investigated by coflocculation of flocculent strains of Saccharomyces cerevisiae, of both Flo 1 and NewFlo phenotypes, to known mnn mutants which vary in the wall mannan structure. Strong coflocculation was found with mnn1, mnn4, mnn9 and control strains, while very little coflocculation was found with mnn2 and mnn5 strains. In contrast, aggregation of these mutants by concanavalin A, a lectin with similar sugar inhibition to NewFlo phenotype flocculation, showed strong aggregation of mnn1, mnn4 and mnn5 strains and poor aggregation of mnn2 and mnn9 strains. The mmn mutant data suggested that flocculation receptors were the outer-chain mannan side-branches, two or three mannose residues in length, confirming an earlier theory based on sugar inhibition data. The similarities and differences between flocculation and concanavalin A aggregation are discussed.


Mutational analysis of Schizosaccharomyces pombe U4 snRNA by plasmid exchange.

We have developed a system for testing mutations by plasmid exchange in the fission yeast Schizosaccharomyces pombe. This system has been used to test the requirement for different regions of the small nuclear RNA U4 in S. pombe. Surprisingly, five of seven deletion and substitution mutations tested in different regions of U4 prevent the accumulation of the mutant RNA. Substitution of the U4 sequence in stem 1 of the U4/U6 interaction domain allows accumulation of the mutant U4, but does not support viability. Two sequences with homology to the Sm binding site are found in the 3' region of S. pombe U4; substitution of the 3' sequence of the two does not interfere with accumulation or function of U4, indicating that the 5' sequence is the functional Sm-binding site.


The telomere-associated MAL3 locus of Saccharomyces is a tandem array of repeated genes.

Saccharomyces strains capable of fermenting maltose contain any one of five telomere-associated MAL loci. Each MAL locus is a complex of three genes encoding the three functions required to ferment maltose: maltose permease (GENE 1), maltase (GENE 2) and the MAL trans-activator (GENE 3). All five loci have been cloned and all are highly sequence homologous over at least a 9.0 kbp region containing these GENEs (Charron et al., Genetics 122, 307-331, 1989). Our initial studies of strains carrying the MAL3 locus indicated the presence of linked, repeated MAL-homologous sequences (Michels and Needleman, Mol. Gen. Genet. 191, 225-230, 1983). Here we report our analysis of the centromere-proximal MAL3-linked sequences and show that the complete MAL3 locus spans approximately 40 kbp and consists of tandemly arrayed, partial repeats of the three GENE sequences described above. In addition, the structure of the MAL3 locus is compared to that of three partially functional alleles of MAL3. These alleles were shown to contain only MAL31 and MAL32 and their structure suggests that they resulted from MAL3 deletions removing the sequences centromere-proximal to MAL31. The amplification and rearrangement of the telomere-linked MAL3 sequences are discussed in the context of studies on other telemere-associated sequences from yeast and other species.


Efficient selection of phleomycin-resistant Saccharomyces cerevisiae transformants.

The recently described dominant yeast marker Tn5ble confers phleomycin resistance on the yeast Saccharomyces cerevisiae (Gatignol, Baron and Tiraby, 1987. Mol. Gen. Genet. 207, 342-348). Incubation in non-selective medium prior to selection is critical, however, for getting phleomycin-resistant transformants. A 6-h incubation period was found to give optimal transformation frequencies, up to 10(5) transformants/micrograms plasmid, comparable to selection for uracil prototrophy (Ura+).


MOL1, a Saccharomyces cerevisiae gene that is highly expressed in early stationary phase during growth on molasses.

We have isolated a new Saccharomyces cerevisiae gene, MOL1, that is transiently expressed at high levels in the early stationary phase of batch cultures growing on industrial molasses medium. The DNA sequence of the MOL1 gene (for MOLasses-inducible) with its flanking regions was determined (EMBL accession number X61669). It encodes a polypeptide of M(r) 35 kDa that is closely related to stress-inducible proteins of similar size from two Fusarium species. Unlike ST135 of Fusarium, MOL1 is not induced by ethanol or heat shock. MOL1 expression is absent in rich (YP) medium, and only very low levels of expression are detectable in minimal (YNB) medium. The gene is not essential, and a MOL1 disruption strain showed no apparent phenotype under a variety of growth conditions. The 5' region of MOL1 contains the complete sequence previously determined for the SUF4 locus, encoding a tRNA-gly (UCC) gene, which has been mapped to chromosome VII.


RHO gene products, putative small GTP-binding proteins, are important for activation of the CAL1/CDC43 gene product, a protein geranylgeranyltransferase in Saccharomyces cerevisiae.

Two multicopy suppressors of the cal1-1 mutation in the yeast Saccharomyces cerevisiae have been isolated and characterized. They are identical to the yeast RHO1 and RHO2 genes, which encode putative small GTP-binding proteins. Multiple copies of either RHO gene suppressed temperature-sensitive growth of the cal1-1 mutant but did not suppress the cal1 null mutant. Genetic analysis suggests that overproduction of either RHO gene product acts for activation of the CAL1 gene product.


Disruption and mapping of IDI1, the gene for isopentenyl diphosphate isomerase in Saccharomyces cerevisiae.

Isopentenyl diphosphate isomerase catalyses an essential activation step in the isoprene biosynthetic pathway. The Saccharomyces cerevisiae gene for isomerase, IDI1, was recently isolated and characterized (Anderson et al. J. Biol. Chem. 1989a, 264, 19169-19175). Wild-type IDI1 was disrupted with a LEU2 marker, and the resulting DNA was used to transform a yeast leucine auxotroph. Southern blots of EcoRI fragments of chromosomal DNA from the diploid strain showed the expected fragments for intact and disrupted IDI1. Dissection and analysis of tetrads demonstrated that IDI1 is an essential single-copy gene. A CHEF gel and clone grid filter analysis, followed by chromosomal mapping indicated that the gene is located on chromosome XVI approximately 55 kb centromere proximal to PEP4.


DNA sequencing and analysis of a 24.7 kb segment encompassing centromere CEN11 of Saccharomyces cerevisiae reveals nine previously unknown open reading frames.

A 24.7 kb segment of the cosmid clone pUKG047 containing a Sau3AI-partial fragment from the centromere region of Saccharomyces cerevisiae chromosome XI was sequenced and analysed. A mixed strategy of directed methods including exonuclease III nested deletion, restriction fragment subcloning and oligonucleotide-directed sequences was carried out. Exclusive use was made of the Applied Biosystems Taq DyeDeoxy Terminator Cycle technology and a laser-based AB1373A sequencing system for reactions, gel electrophoresis and automated reading. A total of 12 open reading frames (ORFs) was found. Nine new ORFs (YK102 to YK110) were identified, three of which (YK102, YK107, YK108) showed homologies to proteins of known function from other organisms. In addition, sequence analysis revealed three recently functionally characterized genes (MET14, VPS/SPO15, PAP1), which could be joined to the earlier published CEN11 region.


Sequence of a 12.7 kb segment of yeast chromosome II identifies a PDR-like gene and several new open reading frames.

A 12,684 bp DNA fragment, between FUS3 and the centromere, from the left arm of chromosome II of Saccharomyces cerevisiae was sequenced as part of the European project to sequence the whole chromosome. This segment contains at least five complete new open reading frames (ORFs) and the beginning (191 first 5' codons) of an ORF whose putative translational product is highly similar to the multidrug resistance PDR1 gene previously characterized by Balzi et al. (1987) on chromosome VII.


An 11.4 kb DNA segment on the left arm of yeast chromosome II carries the carboxypeptidase Y sorting gene PEP1, as well as ACH1, FUS3 and a putative ARS.

We report the nucleotide sequence of an 11.4 kb DNA segment from the left arm of Saccharomyces cerevisiae chromosome II. This sequence contains a typical structure of a functional ARS as well as five open reading frames (ORFs) longer than 300 bp. One is PEP1, a gene encoding a transmembrane protein of 1579 amino acids which transits through the secretory pathway and is involved in vacuolar protein sorting. Two genes were previously sequenced: ACH1 (Lee et al., 1990) and FUS3 (Elion et al., 1990), which encode an acetyl-CoA hydrolase and a protein kinase involved in the cell division cycle, respectively. The last two ORFs localized on the complementary strand of ACH1 are not likely to be expressed.


AFG1, a new member of the SEC18-NSF, PAS1, CDC48-VCP, TBP family of ATPases.

We have sequenced a gene that encodes a 377 amino acid putative protein with an ATPase motif typical of the protein family including SEC18p (NSF = N-ethyl maleimide-sensitive fusion protein; vesicle-mediated endoplasmic reticulum to Golgi protein transfer), PAS1p (peroxisome assembly), CDC48p (VCP = valosin-containing protein; cell cycle) and TBP1 (Tat-binding protein). This gene, AFG1 for ATPase family gene, also has substantial homology to these proteins outside the ATPase domain. AFG1 is located on chromosome V immediately centromere-proximal to MAK10.


Sequence of the genes encoding subunits A and B of the vacuolar H(+)-ATPase of Schizosaccharomyces pombe.

The genes encoding subunits A (vma1) and B (vma2) of the vacuolar H(+)-ATPase from Schizosaccharomyces pombe were cloned by hybridization to cDNAs of the homologous genes in Neurospora crassa. Both genes are interrupted by introns, two in vma1 and four in vma2. Positions of introns do not appear to be conserved when compared to those of N. crassa. The subunit A gene encodes a single product of 619 amino acids and is not interrupted by the coding sequence for a second product as found for Saccharomyces cerevisiae (Kane, P. K., Yamashiro, C. T., Wolczyk, D. F., Neff, N., Goebl, M., and Stevens, T. H. (1990). Science 250, 651-657).


LEU2 gene homolog in Kluyveromyces lactis.

A DNA fragment that can complement the leu2 mutation of Saccharomyces cerevisiae was cloned from the genomic library of Kluyveromyces lactis. The nucleotide sequence revealed an open reading frame of 362 codons, 75% homologous to S. cerevisiae LEU2 gene. The upstream region contained a CCGGAACCGG sequence identical to the site of leucine-specific control of LEU2. Further upstream, there is a partial open reading frame homologous to rat ribosomal protein L7.


[Determination of water-soluble vitamins B1, B2, B6 and B12 in milk using HPLC].

Simple methods of determining the water-soluble vitamins B1, B2, B6 and B12 in milk by HPLC are described. Compared to existing procedures, the following improvements can be realized. The oxidation of vitamin B1 to thiochrome is stopped by the addition of sodium sulphite. This step significantly increases repeatability. Thiochrome is then extracted with butan-1-ol, which results in fewer co-extracts and greater selectivity. After the hydrolysis of the 5-phosphates of the vitamin B6 (pyridoxine, pyridoxal and pyridoxamine), these three vitamers are determined by isocratic HPLC as DDS-ion-pairs and with fluorimetric detection. As only microbiological methods have so far been used for the determination of minute quantities of vitamin B12 in milk, a new HPLC procedure is proposed with a detection limit of 0.2 micrograms vitamin B12/L milk.


A rapid, sensitive and economic method for the detection, quantification and confirmation of aflatoxins.

A rapid, sensitive and economic method for the detection, quantification and confirmation of aflatoxins is described. Aflatoxins B1, B2, G1, and G2, are extracted by methanol/water (85 + 15) and partitioned into methylene dichloride. The methylene dichloride solution is cleaned up on a polypropylene column, filled with 0.5 g silica gel 60. The aflatoxins are eluted with chloroform-acetone (90:10) and are detected using bidirectional thin-layer chromatography (TLC) with aluminium silica gel foil. The mean recovery of aflatoxins B1, B2, G1, and G2 in corn samples was 73, 78, 80, and 64%, respectively; the limit of detection was 0.5 micrograms/kg. The results can also be confirmed by derivative formation using trifluoroacetic acid on the TLC plate. The method has been applied to a wide range of foods with good results.


Rapid analysis of whey proteins from different animal species by reversed-phase high-performance liquid chromatography.

This paper explores the possibilities of reversed-phase high performance liquid chromatography (RP-HPLC) for analysing whey proteins from the milk of cows and other animal species. An RP-HPLC method is proposed to separate and quantify bovine whey proteins. Using this method, bovine serum albumin, alpha-lactalbumin, beta-lactoglobulin A and beta-lactoglobulin B were separated in less than 7 min. It is demonstrated that irreversible adsorption of bovine whey proteins occurs on unused columns (i.e. those not previously used to separate proteins). Therefore, prior conditioning of the column with whey proteins is required for valid protein quantification. Conditioning can be achieved by eluting a large amount of at least one of the bovine whey proteins through the unused column. The calibration curve (peak area vs. protein concentration) for the main bovine whey proteins was linear. This method also allowed good separation of caprine and ovine whey proteins and separation of some homologous whey proteins of different animal species. Detection of milk mixtures from different animal species was carried out using this method.


[Recent advances in neuropsychopharmacology of the central serotonin receptor].

The existence of multiple serotonin (5-HT) receptor subtypes has been proposed based on radioligand binding assay technique and other functional assay. Recent advance of neuropsychopharmacology has contributed to elucidating their physiological functions, ranging from molecular biological to clinical characteristics. Abnormalities of central 5-HT function are currently thought to play a significant role in mental disorders such as affective disorder, anxiety disorder, eating disorder and negative symptoms of schizophrenia, and in the regulation of physical functions such as body temperature, blood pressure and pain. The most significant outcome of the basic pharmacological work has been successful application of 5-HT receptor agents to the treatment of the above clinical disorders. In this article, the authors review the history of 5-HT receptor research and the role of 5-HT receptor in clinical disorders.


[Patch-clamp technique].

We first describe how to set up the system for patch-clamp recording and carry out the experiments with 4 different modes of conventional patch-clamp technique: cell-attached, whole-cell, inside-out and outside-out configurations. Thereafter, we mention about an improved 'nystatin perforated' patch-clamp technique which dissolves the fault of conventional patch-clamp techniques except the cell-attached mode.


Differential effects of intracerebroventricular AF64A injection, nucleus basalis of Meynert lesion, and scopolamine on place navigation and open-field behavior of rats.

The present study was conducted to compare the behavioral effects of intracerebroventricular AF64A injection, electrolytic lesion of the nucleus basalis of Meynert (NBM), and systemic scopolamine treatment, all of which are typical experimental models of central cholinergic system dysfunction, on place navigation performance and open-field behavior in rats. The animals were trained in a circular pool to escape from opaque water onto the hidden platform fixed at a particular location for 5 days. Significant performance deficits were observed in all the models tested. However, it is of importance that any impairments relating to information storage within a day or across days were not obtained. Choline acetyltransferase activity was decreased in the hippocampus of AF64A-injected rats and in the cortex of NBM-lesioned rats, respectively, but those changes did not simply associate with the reductions of the performance. These results suggest that the central cholinergic system is only partially involved in cognitive process. In the open-field test, ambulation, rearing, defecation, and urination were measured for 5 min. Patterns of changes in the open-field behavior were clearly different among the 3 models, indicating variability of the effects of these cholinergic manipulations on motor activities and emotionality.


[Effects of mesocortical dopaminergic lesions on stress-induced changes in monoamine metabolism in the discrete brain regions of rats].

This study assessed the possible influence of the mesocortical dopamine (DA) system on the reactivity of subcortical monoamine systems to stress. Rats were either sham-operated or subjected to DA-depleting lesions of the medial prefrontal cortex (mPFC) with 6-hydroxydopamine. Fourteen days after surgery, the rats were exposed to 30 or 60 min immobilization stress, and levels of noradrenaline, 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG). DA, 3-4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), serotonin, and 5-hydroxyindoleacetic acid were measured in nine brain regions. The stress caused a significant increase in the levels of DA metabolites, DOPAC or HVA, in the mPFC of sham-operated, but not of lesioned animals. HVA concentrations in the nucleus accumbens, olfactory tubercle, and ventral tegmental area were significantly increased in both sham-operated and lesioned rats, but the effect of stress on the subcortical DA systems was unaffected by the lesion. The stress also significantly elevated MHPG levels in the amygdala and hypothalamus of both groups of animals, and these results were similar in the amygdala of rats with or without lesions. In the hypothalamus, however, 60 min stress caused a more marked increase in MHPG concentrations in lesioned rats than in sham-operated rats (lesion-group, 143%; sham-group, 122%), while 30 min stress produced a similar elevation in both groups of animals (lesion-group, 149%; sham-group, 142%). These results suggest that the mesocortical DA system may modulate the sensitivity of the noradrenergic system in the hypothalamus to stress.


[Quality assurance: clarification of a provocative and trendy expression].

The GRG from 1988 commits all suppliers of medical or therapeutic services in Germany to take steps to assure quality. The legislator left it in the care of the involved parties to find an agreement how to do it. This vagueness together with the emphasis put simultaneously on quality and economy turned this issue into a threat for many therapists. This tendency is supported in addition by the intensified competition between the therapeutic schools. The major goal of this review is to give some information that provide a functional discussion and hinder a misuse. The article informs about the issue of quality assurance, which is not as new as some people may consider it; the political context is pointed out, the main concepts are introduced, approaches and experiences from other countries or other fields of medicine are reported, and the conflicts between the interest groups are sketched out. Finally, some comments on central aspects are given which eventually make easier a transference into the field of psychotherapy and hopefully encourage specific approaches of quality assurance in this field.


[The Harald Schultz-Hencke intentionality concept from the viewpoint of psychoanalytic infant research].

Recent Research on small children has infused Schultz-Hencke's concepts of intentionality and the experiencing of drives with new relevance. Because of the fundamental similarities between Schultz-Hencke's work and psychoanalytical infant research, Lichtenstein's theory of motivation is being compared to Schultz-Hencke's experiencing of drives.


[Psychotherapy of depressed elderly patients with pathologic grief reaction].

Loss events not sufficiently worked through frequently are the core syndrome of depressive illness in the elderly. The frequency and symptom pattern of pathological grief reactions as well as influencing factors in 155 patients 60 year old or older treated for endogenous, neurotic and reactive depression, are described. Special issues of the psychodynamics of pathological grief reactions in the elderly are discussed; problems of counter-transference are pointed out. The developmental potential in the elderly often is underestimated. Psychotherapy of pathological grief reactions in the elderly offers important and rarely used possibilities.


[Elderly patients in psychosomatic psychotherapeutic institutions].

With the aid of a comparative study of the data collected in the Berlin Psychotherapy Study and the outpatient clinic of the Psychosomatic Clinic (University of Heidelberg), the allocation and utilization modalities of elderly patients with regard to psychosomatic-psychotherapeutic institutions is investigated. One important result seems to be the high rate of psychodynamically understandable triggering situations (as a measurement of psychological comprehension of the symptoms) and the consequent therapy recommendations (as a measurement of treatability) even for elderly patients. A case study will demonstrate that the danger may arise to subsume new somatic symptoms in the elderly under the aspect "multimorbidity" and thus withhold recommended psychotherapeutic treatment.