[Establishment of a new human renal cancer cell line (TC-1) and its productivity of interleukin-6 (IL-6)]. We established a new cell line (TC-1) from primary site of a renal cell carcinoma (RCC) patient. Its doubling time in tissue culture was 20 hours at 45th passage and mycoplasma contamination test was negative. The karyotypic analysis demonstrated a human karyotype with a modal number of 70. A consistent chromosomal abnormality was noted such as No. 4 monosomy, No. 7 trisomy and a loss of Y chromosome. Electron microscopic examination showed a brush border, vacuoles and abundant glycogen granules in the cytoplasm, which was compatible with RCC cells. This cell line was transplantable to nude mice and the grown tumor closely resembled the original tumor, i.e. clear cell type and hypervascularity. High titer of interleukin-6 (IL-6) was detected in the supernatant of TC-1 cell culture (approximately 5 ng/ml) as well as in sera of nude mice bearing this tumor (260 pg/ml). Exogenous IL-6 did not enhance the TC-1 cell proliferation as determined by cell count. Flow cytometric analysis could not demonstrate the existence of IL-6 receptor on the cell surface. These results suggested the produced IL-6 did not act as an autocrine growth factor in the cell line. Additional IL-1 alpha to the culture medium induced 3-4 times higher concentration of IL-6 in the culture supernatant compared with that of non-stimulating cells, while exogenous TNF alpha did not stimulate IL-6 production.


[Analysis of early and late complications and their origins in Kock continent urinary reservoir following technical modification]. Kock and associates have made a major contribution to the ideal continent urinary diversion by developing a method for creating a continent internal reservoir for urine using the ileum. However, substantial problems have unfortunately resulted due to early and late postoperative complications. In an effort to further elucidate this problem, we herein summarize the incidence of early and late postoperative complications following a series of technical modifications made in patients who underwent Kock continent ileal reservoir construction from our ongoing experience to develop more suitable modifications of this procedure. From January 1985 through December 1991, 47 patients have undergone Kock pouch construction for continent urinary diversion. Our basic technique utilized in this study was slightly modified from that described by Skinner et al. Based on our earlier experience, several changes in the technique were made. One major change is that one row of surgical staples fixes the created nipple to the back wall of the reservoir for the efferent limb, and a further 3-cm longitudinal mucosal incision is made through the outer layer of the intussuscepted ileal nipple and the opposite wall of the reservoir which are sewn to the reservoir edge. A total of eleven patients (11/47) suffered one or more early surgical complications. A 31.8% (7/22) morbidity with an operative mortality rate of 4.3% (2/47) and a 16.0 (4/25) morbidity were observed in the early phase and late phase, respectively. Prolonged urinary leakage from the ileal reservoir and/or through a pin hole in the afferent limb made by surgical staples was observed in six patients (27.3%) in the early phase.(ABSTRACT TRUNCATED AT 250 WORDS)


[A case of Noonan syndrome with neurogenic bladder]. We report a case of Noonan syndrome with neurogenic bladder. The patient was a 33-year-old man with a chief complaint of difficulty in urination, urinary frequency, and incontinence of urine. He had also various malformations, as deformities of the face, skull and hard palate, webbed neck, nail abnormality, edema of the lower extremities, contraction of visual field, deafness, malrotation of the intestine and so on. The case was diagnosed as Noonan syndrome because his karyotype of chromosome was 46, XY. Malformations of the urogenital organs were mild malrotation of bilateral kidneys, a right renal cyst, left cryptorchidism and pseudo phimosis. The uroflowmetrogram of the case showed a saw-like and flat pattern, with urination starting by tapping lower abdomen by himself. The cystometry and electromyogram of the external sphincter revealed neurogenic bladder with uninhibited contraction and detrusor-external sphincter dyssynergia. This is the first case of Noonan syndrome with neurogenic bladder.


[Drug induced cystitis due to ketotifen fumarate--a case report]. Tranilast, an antiallergic drug, is well known as a causal drug of cystitis, and a report is made here of our experience of 1 case of drug-induced cystitis ascribable to ketotifen fumarate. A 13-year-old female had been taking anti-asthmatic drugs orally since the onset of athmatoid attacks, from age of 5. The attacks intensified from the age of 12, because of this she began to take various anti-asthmatic drugs orally. She visited another hospital, due to pollakisuria, in November, 1990, and received treatment for cystitis. However, the symptoms were not alleviated, and she visited our department on January 9, 1991. By urinalysis, large counts of leukocytes and erythrocytes were observed in a visual field of the sediment. Remarkable reddening was observed over the urinary bladder in the patient's cystoscopic findings. Treatment was given at our department, on an outpatient basis, with various antibacterial drugs for approximately one month, but her symptoms were not alleviated, pollakisuria and aseptic pyuria persisted. The patient had never taken tranilast; oral intake of ketotifen fumarate and saibokutou was discontinued on February 13, due to a suspicion of drug-induced cystitis, and her symptoms subsequently disappeared. On February 22, she took ketotifen fumarate orally again, on her own, due to asthmatoid attack, and her symptoms returned. The oral intake of ketotifen fumarate was again discontinued, and alleviation of the symptoms and normalization of the urinary findings were again observed. As a result, lymphocyte stimulation tests on all the drugs the patient had ever taken, only ketotifen fumarate turned out to be positive.(ABSTRACT TRUNCATED AT 250 WORDS)


Role of cyclooxygenase system in cerebrovascular responsiveness: different effects of indomethacin on CO2 responsiveness and dilatation by Ca(2+)-channel blocker. To investigate roles of prostaglandins in the regulation of cerebral blood flow, we compared effects of indomethacin, a cyclooxygenase inhibitor, on the cerebrovascular CO2 responsiveness with those on the cerebrovascular dilatory action of diltiazem, a Ca(2+)-channel blocker. Fifteen adult cats were used. The cerebral tissue oxygen tension, carbon dioxide tension, pH and blood pressure were measured continuously. Indomethacin (1 mg/kg) was infused into the carotid artery. In 8 cats, 3 min inhalation of 5% CO2 in air was performed before and after the indomethacin infusion. In 7 cats, diltiazem (100 micrograms/kg) was infused into the carotid artery for 3 min before and after the indomethacin infusion. The cerebrovascular CO2 responsiveness was significantly decreased (p < 0.05) after the administration of indomethacin. On the other hand, the cerebrovascular dilatation induced by the Ca(2+)-channel blocker was significantly increased (p < 0.05) after the administration of indomethacin. It is concluded that the products of cyclooxygenase system are involved in the cerebrovascular responsiveness both to CO2 and to Ca(2+)-channel blocker, but action mechanisms of prostaglandins may be different, that is, prostaglandians may enhanced cerebrovascular responsiveness to CO2 but diminish it to Ca(2+)-channel blocker.


Studies on contrast visual acuities in normal, cataractous and pseudophakic eyes. Contrast sensitivity and glare sensitivity are often abnormal in cataract patients. However, despite significant subjective complains, relatively good visual acuities are obtained with high-contrast optotypes. Using Variable Contrast Visual Acuity Charts with contrast levels of 90, 15, and 2.5% and reverse polarity of 90% contrast, contrast visual acuities were measured in 24 normal subjects (40 eyes) (Landolt visual acuity > or = 1.0); 28 cataract patients (40 eyes) (Landolt visual acuity > or = 0.6); and 75 patients (100 eyes) [best-corrected Landolt visual acuity > or = 0.8 after intraocular lens (IOL) implantation]. The cataract group experienced the greatest decrease of contrast visual acuity when the high-contrast chart (chart 1) measurements were compared with the 15% (0.71 vs 0.52-0.64 octave) and the 2.5% contrast charts (1.75 vs 1.21-1.48 octaves), followed by the IOL group, and the normal subjects. In the cataract group, 23/40 eyes (57.5%) showed better contrast visual acuity with chart 4 than chart 1. Although much less in frequency, 28/100 (28%) eyes with IOLs also showed better contrast acuity measured with chart 4 than with chart 1. In addition to high-contrast optotypes, acuity measurements using intermediate- to low-contrast optotypes, combined with the reverse polarity chart, seem effective to analyze visual disabilities caused by early cataract. The pattern of the contrast acuity profile of the IOL patients was comparable to normal subjects, but the glare effect still existed after IOL implantation, though to a lesser degree than in cataractous eyes.


"Swimming-induced head twitching" in rats in the forced swimming test induced by overcrowding stress: a new marker in the animal model of depression? We have used overcrowding stress to study the pathogenesis of depression and the action of antidepressant drugs. In the present study, the influence of overcrowding on behavior was assessed by the forced swimming test. All the stressed rats revealed highly characteristic head twitching movement, which was not inhibited by repeated administration of diazepam and haloperidol, but was markedly suppressed by repeated administration of desipramine and mianserine. A significant positive correlation in the number of twitching episodes in each stressed rat between the first and second forced swimming test was seen. These findings support the use of overcrowding of rats as a stressor in the animal depression model because it fulfills the criteria of the model; face validity, construct validity and predictive validity. We propose the adoption of "swimming head twitching" as a new marker in the animal model of depression.


[Primary arterial hypertension. 24-hour recording or casual blood pressure measurement?]. In 134 patients with essential hypertension 24-hour blood pressure ambulatory monitoring (ABPM) as well as traditional blood pressure measurements (casual BP) were performed. Maximal and mean whole-day systolic and diastolic blood pressure values in ABPM were compared with casual BP (using test t). The casual BP was also compared with mean systolic and diastolic blood pressure values calculated separately in three eight-hours periods of day. Maximal BP values in ABPM were significantly higher and mean significantly lower than casual BP values. Blood pressure registered during work hours was closest to casual BP values. Blood pressure in the evening was similar to whole-day blood pressure mean. The results indicate, that automatic blood pressure monitoring performed in this period of day has the greatest diagnostic value because it could be free from "white coat hypertension" syndrome.


[Heparin increases platelet aggregation and inhibits fibrinolysis, especially in primary hypertriglyceridemia and after myocardial infarction]. The authors presented the effect of unfractionated heparin on platelet aggregation and fibrinolysis. The studies have been performed in the control group and with primary hypertriglyceridemia as well as in patients after myocardial infarction. Adrenaline and ADP induced platelet aggregation was examined in plasma rich platelets; fibrinolysis was estimated in euglobulin test. These parameters have been determined before and after (10, 20, 60, 240, 360 min.) intravenous injection of 50 units heparin per kg body weight. We have observed, that heparin increased also platelet aggregation and inhibited fibrinolytic activity in persons with primary hypertriglyceridemia particularly and after myocardial infarction. Correlation was found between investigated parameters and products of heparin lipolytic activity, especially with level of free fatty acids.


[Angioplasty of the ostium of the left coronary artery using a venous patch]. Isolated critical ostial stenosis of the left main coronary artery (LMCA) without narrowing in the distal parts of coronary vessels is rather rare cause of angina. It was observed in 7 our patients: 5F and 2M aged 42-55 yrs (mean 47.5). Five of them were in unstable condition. In all of them a direct surgical angioplasty of the LMCA was performed. Cardiopulmonary bypass with moderate hypothermia were used in all patients. The LMCA was approached from behind. A curved incision was made into the right lateral aortic wall toward the LMCA. Care was taken to stay away from the commissure between the noncoronary and the left coronary cusp. The posterior aspect of the LMCA was incised across the stenosis and prolonged through bifurcation. A venous onlay patch was used to enlarge not only the LMCA but also the adjacent 2 cm of aortic incision, so as to give the LMCA ostium a funnel shape, which favors a homogeneous blood flow. The mean aortic cross clamping time was 46 min. The patients were easily weaned from cardiopulmonary bypass. The early and late results are good--all patients were discharged from the hospital free of symptoms. In 6 patients a perfect patency of the left main stem was documented during control coronarography. In our opinion direct surgical angioplasty of LMCA is better then the conventional surgical treatment because normal geometry of LMCA ostium and normal blood flow can be restored using this method.


[Surgical treatment of ventricular tachycardia in patients with post-infarction aneurysms]. 4 patients (P) with recurrent, sustained ventricular tachycardia (VT) resistant to medical treatment, underwent surgery for cure of this arrhythmia. Each P had episodes of VT lasting 30 or more seconds, 3 of them had episodes of ventricular fibrillation. In all cases rhythm disturbances were secondary to post myocardial infarction aneurysm. Coronary angiography showed in all P total occlusion of LAD, in 2 cases significant lesion in RCA were found. 1 P had lung cancer. All P underwent aneurysmectomy and an excision of the altered endocardium by Harken's method. The endocardial excision was performed without endocardial mapping. 2 P had concomitant CABG to RCA. In the P with lung cancer lobectomy was performed. There were 2 ++non-arrhythmic death. The P with lung cancer died because of sepsis due to lung abscess. One P died because of heart failure (preoperative EF 10%), 6 months after the surgery. The 2 survivors remained free of VT during a follow-up period 8 months. In conclusion, endocardial excision by Harken's method is efficient in treating recurrent sustained VT, resistant to medical treatment, in patients with post myocardial infarction aneurysm. The surgical procedure can be performed without intraoperative endocardial mapping.


[Surgical treatment of bacterial endocarditis of the tricuspid valve]. We present a rare case of bacterial endocarditis of tricuspid valve caused by temporary intracardiac pacing. The 48-year old male patient developed complete a-v block during the 1st day of acute inferior myocardial infarction. Intracardiac electrode was inserted for temporary pacing. After 4 days signs of bacterial endocarditis developed. Patient was markedly febrile, moderate tachycardia with gallop rhythm and systolic murmur of tricuspid valve insufficiency were present. Dullness to percussion was audible at the base of right lung. Hepato- and splenomegaly appeared during the second month of hospitalization. Laboratory tests revealed: elevated ESR, leukocytosis with a shift to the left, several blood cultures were positive to Staphylococcus aureus. On repeated chest X-ray patchy infiltrates with thin-walled translucent pools were visible. Transthoracic and transoesophageal++ echocardiography provided more precise informations. Bacterial vegetations were visualised on the tricuspid valve. Coronary angiography revealed proximal occlusion of the right coronary artery and 75-80% stenosis of the left circumflex artery. Antibacterial treatment guided by blood cultures was begun: vancomycin combined with netilmycin, then tienamycin and diflucan--after 10 weeks treatment was decided to be unsuccessful and the decision about surgical treatment was made. In extracorporeal circulation posterior left leaflet together with granular bacterial growths was excised. Septal and anterior leaflets were found normal. Cultures made of excised tissue were positive for Staphylococcus aureus and subsequent treatment with fluoroquinolones gave satisfactory result. Postoperative echocardiography revealed only small tricuspid valve insufficiency. Coronary by-pass surgery was performed later because of the high risk of simultaneous operation.(ABSTRACT TRUNCATED AT 250 WORDS)


[Isolated congenital tricuspid valve insufficiency--case report]. A case of congenital, isolated tricuspid regurgitation is reported. A 48 year old woman has been admitted because of shortness of breath and progressive fatigue. Although the heart murmur had been heard in her childhood, the first symptoms appeared when she was 25. One year later she underwent a plastic surgery of tricuspid valve. During the following 20 years she remained asymptomatic, until dyspnea and fatigue developed again. On admission she was cyanotic and positive, systolic jugular venous pulse was seen. Atrial flutter was present and no systolic cardiac murmur was heart. The liver was enlarged without peripheral oedema. Echo-Doppler examination and cardiac catheterisation revealed huge right cardiac chambers and significant tricuspid regurgitation. During reoperation a valve consisting of only two cusps was found with extremely dilated valvular anulus. A Bex valvuloplasty was successfully performed. One month later a control echo-Doppler showed the diminution of right ventricle and right atrium with hardly visible regurgitant jet. The pathogenesis and the course of this extremely rare disease have been also discussed in the paper.


Selective purification of two distinct protein kinases (C-kinase and casein kinase II) from the membrane fraction of mouse brain by NED-affinity column chromatography. By means of NED-affinity column chromatography, two distinct protein kinases have been selectively purified from the crude membrane extract of mouse brain. One (designated P-I kinase) was eluted from the column by the buffer containing 5 mM EGTA and the other (designated P-II kinase) was eluted by the buffer containing 0.6 M KCl. The activity of A-kinase was detected in the column passed through fraction. Biochemical characteristics of P-I and P-II kinases corresponded exactly to those of C-kinase and casein kinase II (CK-II), respectively. In addition, immunoprecipitate experiment using anti-CK-II antiserum against the beta-subunit of Drosophila CK-II showed that P-II kinase is identical to CK-II and the 62 kDa cellular polypeptide is associated with the kinase.


Triacetin as food additive in gummy candy and other foodstuffs on the market. The qualitative and quantitative analytical methods were proposed for the simple and rapid determination of triacetin (TAc) in commercial gummy candies and other foodstuffs by gas chromatography (GC), thin layer chromatography (TLC) and infrared spectroscopy (IR). Each extract from the samples was obtained by pretreatment of the foodstuffs as follows: (A) Gummy candy was dissolved in warm water and the solution was extracted with chloroform. The organic (chloroform) layer was separated. (B) Samples (such as ice cream) containing substantial water were mixed with anhydrous Na2SO4 and stirred to sandy appearance and dried. The residue was homogenized with ether, followed by centrifuging, and the organic (ether) layer was separated. (C) Dried samples (such as chocolate and cookie) were smashed, homogenized with ether, and followed by centrifuging, and the organic (ether) layer was separated. (D) Candy was dissolved in warm water and the solution was extracted with ether. The organic (ether) layer was separated. Each organic layer from (A)-(D) was washed with 10% NaHCO3 and evaporated. The residue containing TAc was dissolved in dichloromethane. The extract obtained was subjected to column chromatography on silica gel. The fractions containing TAc were employed in GC with 25% PEG-20M column, TLC, and IR analyses. Recovery of TAc from gummy candy was 99.1 +/- 3.0% and those from other foodstuffs ranged from was 82.1 to 99.4% by GC. Detection limit by this method was 10 ppm. TAc was found to contain at a level as high as 550 ppm in one domestic gummy candy. On the other hand, one imported gummy candy contained no more than 20 ppm of TAc gummy candy.


Studies of aflatoxins in Chiang Mai, Thailand. Aflatoxins become an economical problem in our country due to its contamination in agricultural commodities for export. The toxin may also cause hepatoma and liver diseases in the Thais as well. It is, therefore, necessary to search for and to develop efficient technology to combat and control such dangerous mold and mycotoxins. This paper is a collection of our previous and present studies towards reduction of risk from aflatoxins in foods and feedstuffs. The investigation of mold and aflatoxin contamination in local foods and feedstuffs in Chiang Mai area was made. The inhibitory effect of garlic extract on growth of Aspergillus flavus and its aflatoxin production was demonstrated. Detoxification of aflatoxins by chemicals such as ammonium carbonate, ammonium bicarbonate and ammonium benzoate was also shown. Preventions of toxigenic mold growth and its aflatoxin production by means of some food preservative were reported. Modifications of such effective chemicals were investigated for safety in future application.


[Neoplastic space-occupying lesions of the orbit. I. Review; hemangioma, lymphangioma and embryonal rhabdomyosarcoma]. From 1987 to 1990 182 patients were treated for orbital lesions in the Department of Ophthalmology of the University of Zürich. 37% were tumors (without tumors of the vascular system), 20% inflammatory lesions, 12% lesions of the vascular system and tumors of it, the rest was not diagnosed. On this background the first of two chapters on orbital tumors deals with diagnosis, differential diagnosis, therapy and follow up of the capillary hemangioma, cavernous hemangioma, lymphangioma and embryonal rhabdomyosarcoma. Although ultrasound echography, CT, MRI and angiography have greatly changed the diagnostic work up of orbital lesions, diagnosis is by far not made straight forward but often only after errors. To enhance diagnosis in the future this study is meant to analyse our misinterpretations. In the case of capillary hemangioma only complications justify an active treatment, normally the lesion resolves spontaneously. The infiltration of the upper eyelid with ptosis needs to be treated because of the risk of amblyopia. Our cases demonstrate, that the attempt of treatment with steroids is justified. Embolisation in the region of the a. ophthalmica is quite dangerous. The cavernous hemangioma only needs treatment in the case of compression of the optical nerve and total excision is not mandatory. The lymphangioma, although classified as benign tumor, can only be excised subtotally and with the danger of traumatising important structures of the orbit. The infiltrative growth and tendency of recurrence in fact is dangerous. It is important to diagnose the tumor without biopsy and to wait with surgical treatment as long as possible. The embryonal rhabdomyosarcoma itself is rare but it is the most frequent primary malignant tumor in childhood. Even accounting for being familiar with mimikri-false history of trauma, inflammation, hemangioma-like angiography-the diagnosis is often made rather late. Our cases demonstrate this and also the change of treatment in the last 30 years. Unfortunately the prognosis of the good results of chemotherapy in combination with radiation if necessary is clouded by the occurrence of secondary malignant tumors.


[Clinical and fluorescein angiography changes in patients with central retinal vein occlusion. A unicenter study of 125 patients]. Between January 1986 and December 1989 we prospectively studied 125 patients with central retinal vein occlusion (CRVO). Documented by fundus photography and fluorescein angiography clinical and angiographic findings were analysed. The age of the patients was between 22 and 89 years with a mean of 60 years. 74 (59%) were male and 51 (41%) female. 63 (50.4%) right and 62 (49.6%) left eyes were affected. Arterial hypertension was found in 34 (37.2%) and diabetes mellitus in 18 (14.4%) of the patients. Glaucoma was present in 14 (11.2%). 11 (8.8%) patients had bilateral CRVO. Preretinal neovascularisation was found in 5 (4%), vitreous hemorrhage in 6 (4.8%) and a retinal detachment in 2 (1.6%) patients. Iris neovascularisation at time of first presentation was found in 8 (6.4%) of all patients, related to the number of ischemic type of CRVO in 17.7%. Cystoid macular edema was found in 77 (61.6%), ischemic maculopathy in 24 (19.2%) patients, and a combination of cystoid and ischemic maculopathy in 11 (8.8%). Mean visual acuity was 20/100. A non ischemic type was present in 80 (64%) an ischemic in 45 (36%) of patients. Mean visual acuity in the non ischemic type was 20/60 and highly significant better (p < 0.0001) than in the ischemic type with a mean visual acuity of 20/400 (Mann-Whitney test). The density of intraretinal hemorrhages (p = 0.0005) and type of maculopathy (p < 0.0001) were highly significant related to the ischemia type (chi-square method).


[Orbital involvement in Wegener's granulomatosis]. Granulomas of the orbit can complicate Wegener's granulomatosis (WG). If they compress the optic nerve, blindness of one or both eyes may result. Therefore, early detection and sufficient treatment are important. Computed tomography (CT) and magnetic resonance imaging (MRI) of the orbit and CNS were performed in patients suffering from WG and orbital granulomas (n = 6). The patients were seen interdisciplinary by internists, ENT-specialists, radiologists and ophthalmologists. 12 out of 121 biopsy proven WG-patients showed orbital granulomas during their disease process. Granulomas of the orbit could be best visualized by MRI. However, MRI and CT taken together were most informative. 6 patients with orbital granulomas are presented as case reports, in four of them WG lead to blindness of one eye. The course of orbital granulomas may be chronic progressive or acutely fulminant. Therapy of first choice is a high dosage, long-term immunosuppression with cyclophosphamide and prednisolone. The detection of orbital granulomas in WG is most successful using CT and MRI. In addition to that, the analysis of visual acuity and visual field are the most important parameters in the detection and follow-up of orbital granulomas. A high dosage, long-term immunosuppression is the treatment of first choice, but in rapidly progressive cases an early decompression of the orbit has to be discussed additionally.


[Static and dynamic infrared thermometry and thermography in malignant melanoma of the uvea and conjunctiva]. Thermometry and -graphy prove asymmetries of the circulation. Contact-free thermometry of the cornea is applied to find out whether this method can contribute to differential diagnosis of ocular melanomas. Under standardized conditions the temperature of the cornea was measured in 30 patients with malignant melanoma of the choroid and the conjunctiva and 35 healthy subjects. The instruments were a handpyrometer (HPM, Messgerätewerk, Magdeburg) and the thermovision camera of AGA Infrared System, Sweden, with dynamic recording in colour. In healthy subjects there are no significant asymmetries in temperature under standardized conditions. The temperature of the cornea is elevated in both malignant melanomas of the choroid and the conjunctiva. Thermometry and -graphy of the cornea can contribute to the differentiation of malignant ocular melanomas from other ophthalmological diseases. They are also suited in long term observation of the course of the melanomas.


[Leiomyoma of the iris--a clinicopathologic case report]. A 20-year-old white man demonstrated a 2-3 mm in diameter ball like, grey white vascularized tumor at 12h in the iris stroma adjacent to the sphincter pupillae OD. The tumor was excised by sector iridectomy. Light microscopically the tumor consisted of densely packed spindle-shaped cells with oval nuclei with granular eosinophilic cytoplasm. Electronmicroscopically numerous 9nm-filaments in parallel configuration were found, fusiform dense formations, basal membranes and pinocytotic vesicles. Positive immunohistochemical reaction for Actin, Desmin and Vimentin, negative for S100.


[A family with blepharophimosis, ptosis, epicanthus inversus and telecanthus. Occurrence of the hereditary marker in five generations]. The condition with blepharophimosis, ptosis, epicanthus inversus and telecanthus is reported in one family over five generations. The syndrome is transmitted as an autosomal-dominant characteristic with a very high penetrance and expressivity, preferentially affecting and being transmitted by males. Affected females were infertile. There were no other accompanying systemic disorders. A three-year-old boy, who develops deprivation amblyopia, was first treated by levator attachment according to Friedenwald's method. Medical canthal surgery followed two years later to correct the epicanthus and telecanthus. A good clinical result was achieved by performing Mustardè's Z-plasty and shortening of the medial canthal ligament. Alternative techniques are reviewed briefly.


[Severe occlusive bilateral retinal vasculitis within the scope of seronegative systemic lupus erythematosus]. Retinal vascular manifestation is the most common form of ophthalmic involvement in patients with systemic lupus erythematosus (SLE). Most frequently these consist of cotton-wool spots with or without intraretinal hemorrhages. Although rare, a more severe retinal vaso-occlusive disease, termed retinal vasculitis, has been described. We report on a 37-year-old white female with a 13-year follow-up of chronic discoid lupus erythematosus, which suffered massive bilateral visual loss coincident with the systemic exacerbation of her disease (proteinuria, pneumonia, serositis, leucopenia). The diagnosis of SLE was established with reference to the revised ARA-criteria (American Rheumatism Association). Ophthalmoscopy and fluorescein angiography revealed the typical aspect of a SLE-associated vaso-occlusive retinopathy on both eyes with marked ischemia of the macula. Immediate maximal immuno-suppressive therapy, early performed panretinal photocoagulation and subsequent cryoretinopexy did not stop the progression of the disease. Six months after the initial event vascularisations of the disc and rubeosis iridis occurred, but no secondary glaucoma up to date. In this patient, the almost complete absence of characteristic autoantibodies and immunological markers was striking. The correlation with other lupus manifestations, different therapeutic concepts and prognostic factors in SLE-associated retinal vasculitis are discussed.


[Neurometabolic diseases. References for classification, clinical aspects and diagnostic procedures]. Diseases caused by neuro-metabolic or neuro-degenerative disorders often lead to diagnostic difficulties, because of their rarity and numerous quantity. The present report tries to combine older classifications according to anatomic aspects with the newer ones underlying biochemical-functional points of view. Then the most important cardinal symptoms in a synoptic demonstration are put in relation to the most considerable neuro-metabolic disorders. Tables concerning age-related diseases and informations on diagnostic methods of importance are presented.


[Reduction of body height in tall girls by estrogen treatment]. Twenty-six girls with constitutional tall stature were treated with 0.5 mg ethinyloestradiol daily po and 10 mg norethisteronacetate from day 21 to 25 of the cycle for 1.4 +/- 0.4 years. Height, weight, bone age according to Greulich-Pyle (GP) and Tanner (RUS), height predictions according to Bayley Pinneau (BP) and Tanner Whitehouse (TW) were recorded at the onset of therapy, at the end of treatment and 7.4 years later. The mean chronological age at start of treatment was 12.6 +/- 1.1 years, bone age (GP) 12.4 +/- 0.7 years, (RUS) 13.5 +/- 0.7 years. The initial height prediction according to BP was 186.2 +/- 4.1 cm, which was 3.5 cm higher than the prognosis according to Tanner Whitehouse. Depending on the method used for height prediction, the height reduction by treatment was 4.6 +/- 3.0 cm (BP) or 1.4 +/- 2.2 cm (TW), respectively. The reduction of predicted height was more pronounced in girls with a bone age (GP) < 12.5 years at the initiation of therapy (n = 13), than in girls with a bone age > or = 12.5 years (n = 13). Final height was measured 7.4 years after the end of treatment. Chronological age: 21.3 +/- 3.7 years, final height 181.8 +/- 3.7 cm. Mean final height after therapy was 7.7 cm above target height, or, after allowance for the secular trend, 3.8 cm above target height. Body weight, recorded in centiles of body mass index, increased from the 38.5 +/- 26 to the 58.1 +/- 22 centile, weight gain was more pronounced in younger girls. At follow up investigation 18/19 girls, who answered a questionnaire, had a positive view of the previous treatment.


Topical ocular anesthetics in ocular irritancy testing: a review. The routine use of topical anesthetics to alleviate discomfort associated with in vivo ocular irritancy testing has been advocated. This review provides information about the adverse effects of topical ocular anesthetics and answers the questions: are topical anesthetics practical and effective in ocular irritancy protocols, is long-term use contraindicated, will topical anesthetics alter the response of a test substance, and are there significant side-effects which might cause pain and suffering in test animals? There was no evidence to support the use of a specific topical anesthetic. Further, information about using systemic analgesics or combinations with local anesthetics that would effectively alleviate discomfort associated with ocular irritancy testing without affecting test results was not found. Comprehensive studies are needed to identify the most effective combination of drugs that would ameliorate discomfort associated with ocular irritation testing.


Comparative study of long, segmented, filamentous organisms in chickens and mice. No transmission of long, segmented, filamentous organisms (LSFO) from a laboratory mouse strain (Mus musculus) to a domestic broiler chicken strain (Gallus domesticus) occurred by oral dosing with mouse mucosal and fecal preparations, even in hydrocortisone-treated chicks. Transmission of chicken LSFO to laboratory mice was attempted, but the results were confounded by the mice having been already infected with LSFO. However, the morphologic characteristics of the LSFO in the dosed mice suggested that transmission of chick LSFO did not occur. Mouse LSFO, as detected in carbol thionin-stained mucosal smears, are generally wider, longer, and have slightly different morphologic characteristics than chicken LSFO. The results of these studies suggest that the LSFO in mice and chickens may be two distinct species.


[Parenteral nutrition in treatment of short stature in adolescents with Crohn disease]. Growth retardation and delayed puberty occur in 20-35% of children and adolescents with Crohn's disease. Alternate day corticosteroid treatment, use of azathioprine, enteral or parenteral hyperalimentation and surgery have been advocated to reverse growth failure. Because of nonacceptance of elemental diet 7 patients with Crohn's disease and growth retardation received parenteral nutrition for 2-3 months (maximal for more than 30 months in one patient). All of them exhibited a mean weight gain of 10 kg and a mean increase of their height velocity from 2.4 to 7.1 cm/year. Main problems were bacterial infections and dislocations of the central lines. Surgery was performed in 3 adolescents immediately after parenteral nutrition. One patient showed a catch-up growth during a 30-months nocturnal home parenteral nutrition at a biological age of 21 years. Parenteral nutrition is an effective regimen to manage growth failure in children with Crohn's disease, but has to be performed for larger periods in individual cases.


Diarrhea caused by a slow-growing Enterococcus-like agent in neonatal rats. An enteropathogenic Enterococcus-like agent was isolated from a spontaneous outbreak of diarrhea that occurred in a colony containing neonatal rats. Diarrhea was experimentally reproduced in virus-antibody-free neonatal rats inoculated with this purified "enterococcus." Gram-positive cocci were adhered to the small intestinal villi of affected animals from which the organism was reisolated. The isolate's classification in the genus Enterococcus was confirmed by genetic probe; however, because of its unique fermentation pattern, it could not be definitively speciated. Indirect immunofluorescence assays indicate that this strain of enterococcus and Enterococcus hirae, another strain pathogenic for neonatal rats, differ antigenically. Enterococci should be considered as potential etiologic agents in outbreaks of diarrhea involving neonatal rats and future efforts directed to increasing our understanding of the pathophysiology of this disease.


[Hereditary heterozygote factor VII deficiency]. Hereditary Factor VII deficiency is one of the rare congenital coagulopathies. Prolonged prothrombin time (PT) with normal partial prothrombin time (PTT) may be an indicator for Factor VII deficiency. A family with hereditary heterozygous Factor VII deficiency is presented in whom no symptoms of a bleeding disorder were clinically detectable. A discussion of the therapeutic options follows.


Nonconvulsive electrocorticographic paroxysms (absence epilepsy) in rat strains. Rat strains were screened for evidence of unresponsive periods associated with high-voltage spike and wave paroxysms on electroencephalography--a rodent model of human absence epilepsy. Five commonly used strains were newly noted to express the spike and wave phenomenon. Only an inbred Sprague Dawley rat strain did not exhibit such episodes. The existence of the phenomenon in many and unrelated inbred rat strains suggests that both genetic and environmental factors are causal.


Tumor dormancy and the effect of selected drugs on the tumor-dormant state. An animal model for studying tumor dormancy was established by two-way selection of tumor-progressive and nonprogressive (tumor-dormant state) ddY mice in the same basal stock. In the tumor-progressive (prg) substrain of mice, Ehrlich ascites tumor cells (2 x 10(7)) subcutaneously inoculated into the dorsal skin formed a progressive solid tumor. In the tumor-dormant substrain (drm) of mice, the same tumor cells did not grow at all but formed a small caked nodule (1 to 3 mm in diameter) within 1 week. Some of the tumor cells persisted in the nodule at least 3 months without apparent mitotic figures. Such dormant tumor cells emerged and revealed outgrowth to overt solid tumors after they were transplanted into the dorsal skin of prg mice or into athymic mice (C57BL/6J nu/nu). To estimate the predisposition of drm mice to form tumors, the effect of certain drugs on the tumor-dormant state was examined. Estradiol, progesterone, dexamethasone, prostaglandin E2, Cyclosporin A, and collagenase all failed to promote the emergence of dormant tumor cells in drm mice.


[Case studies of the effect of tyrosine administration in children with phenylketonuria on cognitive processes]. Eight patients with phenylketonuria and low protein nutrition were treated in a double blind crossover study for the time of 3 month with 150 mg tyrosine per kg body weight and day and placebo thereafter or vice versa. The concentration of phenylalanine in serum was not influenced by the administration of tyrosine whereas tyrosine in serum markedly increased. Psychological tests were 7 times repeated in monthly intervals. From the study it is to suggest that there is an improvement of the test results caused by an effect of training due to the repeated tests and an additional improvement caused by tyrosine.


Preferential alcoholic embryopathy: effects of liquid diets. Our initial report of a preferential expression of experimental alcoholic embryopathy affecting the male offspring contiguous in utero to male siblings of Long-Evans rats was based on gavage administration of alcohol to pregnant rats without regard to isocaloric, pair-fed exposure paradigms. In this study, pregnant Long-Evans rats were given 35% ethanol-derived calories (EDC) in one of two different liquid diets: 1) a liquid alcohol diet based on Sustacal, a flavored liquid food formulated for human nutritional standards; and 2) a high-protein liquid rodent diet devised by Lieber and DeCarli (L&D). The diets were administered from day 6 to 15 of gestation. Pregnant rats were pair-fed liquid diets containing 0% EDC, but isocalorically balanced to 35% EDC with either sucrose (Sustacal) or maltose-dextrin (L&D). A fifth group of pregnant rats was given access ad libitum to standard certified laboratory rodent diet and served as free-fed controls. On day 20 of gestation, all pregnant rats were euthanized and the products of conception examined by standard teratologic techniques. Pregnant animals fed Sustacal-based diets consistently consumed fewer calories per kilogram body weight per day from day 6 to 15 of gestation (i.e., they were significantly calorie-deprived during pregnancy) as compared with the standard laboratory-diet-fed controls or those consuming L&D diets. Body weights of rats consuming Sustacal diets (both 0 and 35% EDC) were significantly lower throughout gestation when compared with all other groups. Higher (> 150 mg/dl) blood alcohol levels were attained by rats consuming 35% EDC in Sustacal diet as compared with L&D (100 mg/dl) diets.(ABSTRACT TRUNCATED AT 250 WORDS)


[Atypical form of long-QT syndrome--a case report]. An unusual case of familial Long QT syndrome is reported. After a longstanding symptom free interval this eight year old girl experienced multiple episodes of malignant ventricular arrhythmias including ventricular tachycardias of the torsades-de-pointes type and ventricular flutter. The treatment with beta- and alpha-blocking agents, phenytoin, calcium antagonists, clonidine and magnesium in increasing doses and varying combinations was unsuccessful. A left sided cardiac sympathectomy failed also to control the life threatening ventricular arrhythmias and to decrease the relative high mean heart rate. Finally a consecutive right cardiac sympathetic denervation in combination with an experimental class I antiarrhythmic agent led to a subjective and objective improvement in the patient's condition which allowed discharge from hospital. Despite a symptom free interval of four months and nearly normalized Holter ECG results the longterm prognosis remains unclear. This unusual case discloses interesting features, because 1) it supports the theory of an intracardiac cellular anomaly rather than the imbalance hypothesis, 2. chronic beta-blocker treatment possibly resulted in an up-regulation of beta-adrenergic receptors which made beta-blocking agents ineffective despite high doses, and 3. an consecutive right sided cardiac sympathectomy resulting in bilateral cardiac sympathetic denervation had to be done-an intervention which is seldom performed in the Long QT syndrome.


Effects of chronic blood loss in a marsupial (Monodelphis domestica). Monodelphis domestica, the gray short-tailed opossum, is used increasingly as an animal model in studies that require repeated blood sampling. Consequently, it is important to establish safe bleeding regimens. We investigated the effects of repeated blood loss on various hematologic values and on different organs in this species. Approximately 2 ml of blood were taken weekly from 20 animals for 13 weeks. The animals were then necropsied; members of an age- and sex-matched control group were bled (2 ml) once and necropsied immediately to obtain baseline data. Ultimately, each animal in the experimental group lost approximately three times its total blood volume. After the first bleeding in the chronically bled group, the red blood cell counts, hemoglobin, and hematocrit values decreased significantly but remained constant thereafter. In another experimental group bled only once, the hematologic values rose to higher than baseline levels after a rest of 2 weeks. Thereafter the values slowly returned to baseline levels. A notable increase in Howell-Jolly bodies occurred in the chronically bled group. Histologically, there was marked erythroid hyperplasia in the bone marrow and extramedullary hematopoiesis in the spleen, but none in the liver. Because there were no obvious detrimental physiologic effects, we conclude that M. domestica is markedly tolerant of chronic blood loss.


[Comparison of the ferritin and iron concentration in capillary and venous blood serum of children, adolescents and adults]. The ferritin and iron concentration were compared in sera from capillary and venous blood (n = 52). Ferritin was determined using a luminescence-enhanced enzyme immunoassay and iron by the ferrozine method. The results showed a very good correlation and a linear relationship for ferritin (r = 0.999) but not for iron (r = 0.855). It is concluded that capillary blood serum can be used only for the determination of ferritin.


[Combination of scimitar syndrome and horseshoe lung. A rare but typical finding--case report and review of the literature]. The scimitar syndrome is a rare congenital cardiopulmonary malformation. In association with a horseshoe lung it is extremely uncommon. We describe a child with typical scimitar syndrome (anomalous right pulmonary venous return, hypoplasia of the right lung with dextro-position of the heart and aberrant systemic arterial supply of the right lung), associated with a horseshoe lung. No clinical symptoms are present, surgical management is not necessary. This is the 20th case in addition to the 19 reported cases with scimitar syndrome associated with horseshoe lung. Eight other additional cases of horseshoe lung without scimitar syndrome are mentioned here. In horseshoe lung the pulmonary parenchyma extends from the right lung base across the midline and fuses with the left lung. In only 3 cases continuous parenchymal tissue could be found histological. In 8 cases the lungs were separated by fissurelike structures, seen in CT or chest film, and in 5 cases fissures were demonstrated histological. From 6 patients no findings are available. The various symptoms, the diagnostic tools and the therapeutic procedure will be discussed.


[McCune-Albright syndrome in association with meningioma and mental and psychological retardation]. Report of a boy aged 17 years and review of the literature on the neoplasias of CAS. The cause of examination was a psychic maldevelopment. Cranial arteriography exhibited extreme wide Carotis communis and CT temporal meningioma right and septum pellucidum cyste. A psychic frontal lobe syndrome and cosmetic impairment by skull changes of the CAS are discussed being partial causes of the psychic maldevelopment. Atypical there were retardation of calcification and dentition, bilateral coarctation of the auditory passage with resulting hearing loss. Till now only J. Frankel and coworkers have published the case of a 50 year old Pakistani with the combination of CAS and Meningioma as a complication of fibrous dysplasia of the skull. We suppose relations between CAS and neurofibromatosis and its blastomatous tendency to meningioma.


Behavioral and physiologic effects of inapparent wound infection in rats. There is a common notion that rats are resistant to postoperative wound infection because many recover from surgery performed under nonsterile conditions. As a result, nonaseptic surgical techniques are used commonly in rat surgery. Our aim was to determine if these techniques cause wound infection and, if so, whether or not the infection, inapparent to casual observation, creates measurable changes in rat physiology and behavior. Rats subjected to craniotomies or laparotomies and inoculated with 10(8) Staphylococcus aureus or Pseudomonas aeruginosa or sterile saline were tested for open-field activity, freezing behavior, home-cage behavior score, and wheel-running activity. Physiologic indices included lactate dehydrogenase, blood glucose, plasma fibrinogen, complete blood counts, wound bacterial counts and histology scores, body temperature, and body weight. Although no clinical signs were detected by postoperative observation, rats inoculated with bacteria were significantly less active in the open field and the duration of freezing behavior was shorter. Plasma fibrinogen, serum glucose, total white blood cell counts, and wound histology scores were significantly altered in the bacteria-inoculated rats. These findings underscore the need for sterile techniques in rat surgery to avoid confounding experimental data.


Effects of isoflurane anesthesia on glucose tolerance and insulin secretion in Yucatan minipigs. Isoflurane's effect on intravenous glucose tolerance and insulin secretion was studied in six Yucatan minipigs. Unanesthetized animals, with previously placed indwelling venous catheters, were tested while resting comfortably in slings. The same animals were then retested during isoflurane anesthesia. Serum glucose and insulin concentrations were measured at predetermined times in response to an intravenous bolus of dextrose. The glucose disappearance rate (k), baseline plasma insulin concentration, the area under the insulin response curve, and the insulinogenic index were significantly lower in the anesthetized animals than in controls. The results of this study indicate that anesthesia with isoflurane significantly alters the glucose/insulin response to an intravenous glucose tolerance test and, therefore, is unsuitable for studies when glucose tolerance is to be assessed.


Cardiovascular effects of a ketamine-medetomidine combination that produces deep sedation in Yucatan mini swine. Seven chronically instrumented Yucatan minipigs were deeply sedated with the combination of ketamine (10 mg/kg), a dissociative anesthetic, and medetomidine (0.2 mg/kg), an alpha 2-adrenoceptor agonist used as an animal sedative in Europe. Both drugs were drawn in the same syringe and administered in the left atrium via a previously inserted permanent catheter. As a result, hypertension (mean arterial pressure from 116 +/- 12 mmHg to 142 +/- 18 mmHg) occurred and was followed by bradycardia (from 107 +/- 22 bpm to 71 +/- 9 bpm). Concomitantly, both the rate of increase in ventricular pressure (48%) and ventricular wall thickening fraction (37%) decreased, thus indicating some worsening of left ventricular function. Further, systemic vascular resistance increased (290%) resulting in a reduction in cardiac output from 0.4 +/- 0.3 l/minute. Also, left ventricular end diastolic pressure initially increased (maximum 10.2 +/- 10.8 mmHg) but returned to the control level in 5 minutes. In spite of an increase in respiratory frequency (3x), PaCO2 increased and PaO2 and pH declined. Rectal temperature decreased from 38.4 +/- 0.9 to 36.0 +/- 0.8 degrees C. All of these changes were transient and returned to control levels during the follow-up period (2 hours). However, epinephrine concentration was exceptionally decreased by the drugs and stayed under the detection limit (20 pg/kg) for the entire time, whereas norepinephrine was undetectable for 10 minutes postadministration. Ketamine-medetomidine, administered in a dose that produced deep sedation, induced marked but reversible changes in most of the cardiovascular variables; there were no pedal or palpebral reflexes for 30 minutes.


Relationship of functional residual capacity to various body measurements in normal sheep. Functional residual capacity (FRC) was determined by nitrogen washout in 55 normal sheep. Data on various external body measurements were collected which included body weight, chest circumference, chest width, body length, height, and sternum length. In addition, data on wet lung weight and wet lung weight/body weight ratio were collected on 10 of the sheep. A significant correlation was found between FRC and all measured parameters except height and sternum length. Multiple linear regression of all external body measurements showed the best correlation of FRC to body weight and body length, while the addition of chest circumference and/or chest width did not significantly improve the correlation. Significant deviation from the population was noted in three sheep (5.5%) that had lung weight/body weight ratios which were significantly lower than the rest of the population.


The selection of an adjuvant emulsion for polyclonal antibody production using a low-molecular-weight antigen in rabbits. Although Freund's adjuvant has been used for decades as an immune enhancer in rabbits, adverse physiologic side effects have prompted the search for more suitable alternatives. We used osteocalcin, a bovine bone protein (M.W. 5,800), as the test antigen to evaluate four adjuvant regimens: a) primary inoculation with complete Freund's adjuvant (CFA) followed by three boosts with incomplete Freund's adjuvant (IFA), b) four serial inoculations with RIBI MPL+TDM+CWS adjuvant, c) four serial inoculations with TiterMax #R-1, and d) primary inoculation (only) with TiterMax #R-1. The antibody yield associated with the CFA/IFA regimen (mean OD = 2.152) was at least sixfold that of either TiterMax (mean OD = 0.358) or RIBI (mean OD = 0.239) multiple injection regimens. No antibody response was observed after the single injection of TiterMax antigen emulsion. Maximal antibody production occurred rapidly in response to Freund's adjuvant (day 31) as compared with TiterMax (day 74) and RIBI (day 66).


Chronic catheterization of the inferior vena cava in Yucatan miniature swine. Long-term venous access for leukapheresis, repeated blood sampling, and administration of drugs and fluids can be accomplished nonsurgically in Yucatan miniature swine. The catheter is placed under fluoroscopic guidance into the inferior vena cava using a needle and guidewire. This procedure has the advantage that it avoids a surgical incision, allows high flow rates, exists conveniently on the lower back, and can be replaced easily in the event of mechanical failure or thrombosis. Actuarial analysis of the duration of patency disclosed that of 41 catheters placed in 30 animals, the probability of function at 28, 42, and 54 days was 75%, 50%, and 25%, respectively. Eleven nonfunctioning catheters were replaced and nine of these continued to function until the completion of the experiment. No catheters were removed due to infection. Chronic catheterization of the inferior vena cava is a convenient method for long-term venous access in swine.


Selecting empiric drug therapy for pneumonia in geriatric patients. Rather than produce extensive lists of pathogens and antibiotics to cover every situation, we have tried to provide some insight into how we make decisions regarding the drug therapy of our geriatric patients when they develop pneumonia, and give some practical examples of how we would use those agents.


Falls in the elderly.

Falls in the elderly. Increasing age leads to an increased risk for both fall and fracture. Debility sufficient to result in institutionalization may double that risk. Multiple risk factors have been identified which are clinically useful in defining individual patient's potential for fall and careful history and physical examination is sufficient to identify most of those risk factors. While falling may be an unavoidable part of a lifetime human experience, the risk of falling should be reduced and its secondary morbidity lessened with a preventive and functionally oriented approach. That approach requires minimal additional resources, but is likely to provide tremendous benefit for both individual patients and the larger society.


Children of alcoholics in the classroom: a survey of teacher perceptions and training needs. Children of alcoholics (COAs) represent a population at risk for alcoholism as well as other physical, emotional, and social problems. This survey explored perceptions of North Carolina elementary and middle school teachers regarding roles and responsibilities in assisting COAs, their need for additional training specific to COAs, and their perceptions of school-based resources for helping COAs. Survey findings teachers generally are willing to assist COAs but need training regarding these children. Training is particularly appropriate for teachers who perceive a low incidence of COAs in their classrooms since these individuals' responses suggest the existence of barriers to effective COA assistance.


Adolescents' perceptions of relative weight and self-reported weight loss activities. Data from the 1990 North Carolina Youth Risk Behavior Survey (YRBS) were used to examine adolescents' perceptions of relative weight and the relationship of these perceptions to physical activity levels, efforts to lose weight, and time spent viewing television. A total of 3,437 ninth and 12th grade respondents to the May 1990 survey were included in the analysis. Of this sample, 25% perceived themselves as "too fat." Of that group, 68% were trying to lose weight. Females made up 75% of those reporting they were "too fat" and were trying to lose weight. White females were more likely to think of themselves as overweight and were more likely to be trying to lose weight than black females. Adolescents who perceived themselves as "too fat" reported fewer days of strenuous activity, fewer hours of strenuous exercise in physical education classes, and more hours spent viewing television on school days.


Culturally relevant smoking prevention for minority youth. A rap contest methodology for smoking prevention was tested with sixth and seventh grade students in a predominantly minority public school district. Contests were held after initial assemblies in which students heard anti-smoking rap messages from same-age and older peers on audio and video tape. Pretesting and posttesting (N = 268) indicated high preference ratings for most aspects of the intervention. Analyses of variance revealed no differences across races on any of the dependent measures. However, smaller assemblies were more effective than larger ones in enhancing attitudes against smoking and obtaining more positive contest evaluations and predictions about smoking behavior. A rap contest method therefore may be effective against the initiation of smoking by disadvantaged children in sixth and seventh grades because it is highly acceptable and perceived as culturally relevant. This effectiveness may be more demonstrable in single classrooms rather than larger assemblies.


Impact of the Nutrition for Life program on junior high students in New York State. School-based nutrition education represents an important component in a national strategy for health promotion. This study evaluated the impact of a school-based nutrition teaching program, Nutrition For Life, on the nutrition knowledge, attitudes, and self-reported behavior of seventh and eighth grade students from New York State. Some 1,863 students in 103 randomly selected classes completed a paper and pencil test covering nutrition attitudes, behavior, and knowledge. Modest use of the teaching program (median use = 3 hours) was associated with modest but significant differences in nutrition attitude, behavior, and knowledge scores. Interactions among teacher assignment, nutrition teaching, and Nutrition For Life use explained significant variations in nutrition test scores. Additional exposure to the program was associated with significantly higher nutrition attitude and behavior scores in schools with a higher proportion of low-income students.


The Midfield High School Safety Belt Incentive Program. Motor vehicle accidents constitute the leading cause of death among children and youth ages 15-24. The Year 2000 Health Objectives for the Nation call for reduction in fatalities in this population through increasing occupant restraint measures. This article provides an overview of how one school district developed a school and community safety belt incentive program to increase safety belt use among high school students. The intervention program followed a seven-step process, involving community leaders, school officials, students, and local law enforcement personnel in its design, implementation, and evaluation. Local merchants and the Alabama Dept. of Health donated program incentives. Measurement of safety belt use among the target population revealed increased use throughout the program.


A formative evaluation of the American Cancer Society Changing the Course nutrition education curriculum. A formative evaluation was conducted of Changing the Course, a behaviorally oriented, activity-based nutrition education curriculum for elementary students from the American Cancer Society, to assess feasibility of program implementation. Sixteen teachers in six schools in the Northeast taught the 15-16 session lower and upper elementary curricula to 702 students. Teacher satisfaction with the curriculum was high. Fidelity of implementation of the curriculum also was high and relatively few modifications to learning activities were made. Teachers reported the one day of training they received was adequate. They also reported a positive influence of the curriculum on themselves and their students. Student achievement results showed most children achieved most of the learning objectives at posttest--80% giving the correct answers on more than 75% of the test items. An examination of children's item-by-item answers on the posttest and of teachers' logs provided information useful for the final stages of development of the curricula and teacher training procedures, before national dissemination.


Promoting health through a developmental analysis of adolescent risk behavior. Adolescents are the only U.S. population group not to improve morbidity and mortality since 1960. Social and behavioral etiologies of adolescent morbidity and mortality are best addressed in schools, since health care providers rarely are seen by teen-agers for other than organic health complaints. The relevance of physical, cognitive, and psychosocial development for adolescent risk-taking and their attendant health problems are discussed. Recognizing adolescent risk-taking as a means of achieving autonomy and identity, nurses and health educators can intervene to promote health-enhancing alternatives to achieve those ends. Social learning theory provides a basis for interventions to personalize the curricula, change behaviors, and shape the environment.


HIV education and health education in the United States: a national survey of local school district policies and practices. To determine the extent to which HIV education and health education policies and practices are required by school districts in the United States, a national probability sample of public school districts was surveyed by mail in 1990. Of 2,150 districts selected, 78.1% responded. HIV education was required by 66.9% of districts. Of these, the percentage requiring HIV education increased by grade level from 29.7% in kindergarten to 82.3% in 7th grade, then declined to 37.3% by 12th grade. Districts that required HIV education most often addressed HIV-related prevention skills in the upper grade levels. Similar to requirements for HIV education, health education requirements also declined from 7th to 12th grade, reaching even lower levels than HIV education by the last two years of high school. These declines are of particular concern given that students are most likely to engage in risk behaviors when HIV and health education is least likely to be required. Other practices and policies that support HIV and health education also were lacking in many districts.


Dietary risk assessment for cardiovascular disease among central Maine adolescents. Dietary risk for cardiovascular disease was assessed in 97 adolescents enrolled in health classes in two high schools in rural, central Maine, using three-day food records and written questionnaires of self-perceived food practices and cardiovascular nutrition knowledge. Mean percent of kilocalories from fat was 36% for both males and females. Compared to American Heart Association guidelines, 80% males and 73% females had more than 30% kilocalories from fat; 37% males and 16% females had dietary cholesterol intakes above 300 milligrams. More than 50% of males but less than 25% of females had sodium intakes greater than 3,000 milligrams. Mean dietary fiber intake of females (6 +/- 4 grams) was significantly lower than the fiber intake of males (11 +/- 8 grams). During adolescence, greater emphasis should be placed on modifying food behaviors that may be detrimental to heart health if continued into adulthood. Individualization provided within nutrition education curricula is necessary to meet the needs presented by differences in gender and variation among adolescents.


[Evaluation of the results of the research funded by the Health Research Fund in 1988]. To analyse the scientific product of research projects funded by Fondo de Investigación Sanitaria in 1988 emphasizing its relation to money granted. 270 out of 610 projects were evaluated in relation to the amount granted. The number of papers published to each project and the impact factor assigned to the journals where these papers were published; we also assessed the mean cost of papers and impact factor units. These projects were coded following UNESCO classifications, and papers as per ISI standards. A total of 95 projects out of the 270 analysed produced no papers; the other 175 projects yielded 471 articles (2.7 per project); the mean cost of each article was 1.1 million pesetas, or 0.8 million if only the productive projects were considered. These papers reached a total of 818,709 impact factor units; the mean cost of the impact factor unit is 660,796 pesetas, or 459,626 pesetas if only productive projects are considered. Non productive projects were those that received less funding. 33% of projects produced papers published in journals with an assigned impact factor equal or inferior to one. There are evident differences among areas of knowledge in terms of impact factor. This study must be completed with a statistical analysis of reported data.


[Inguinal hernia repair modified by Kirschner. A critical analysis after 11 years of clinical experience]. In a retrospective study the early and late complications and the recurrence rate of inguinal hernia 10 years after elective repair by a modification of Kirschner's procedure are presented and compared with those following other established surgical techniques. Questionnaires were sent to 1400 patients, and 1029 patients (73.5) also underwent clinical examination. With all techniques, the most frequent postoperative complications were haematomas and seromas (10.1%). Postoperative mortality was 0.2%. Late complications were reported by 36.4% of the patients followed up by questionnaire. The most frequently reported symptom (by 23.8%) was transitory hypaesthesia in the scar area. In 2.7% of the male patients examined testicular atrophy was found. All patients who reported a recurrence on their questionnaire underwent follow-up examination in the clinic. Thus, the cumulative recurrence rate was 9.6%. Over two-thirds of all recurrences were lateral recurrences (P < 0.01). It was not possible to determine potential risk factors for a recurrence of inguinal hernia. The recurrence rate increased with time before follow up: among the patients examined after 5 years the recurrence rate was 5.7%, while it was 12.1% for patients examined after a period of 10 years. Kirschner's modification involves a risk of lateral recurrence that should not be underestimated, and its use should be reconsidered. The Shouldice repair appears to be the method of choice. However, further results confirming this choice should be awaited.


[Laparoscopic laser cholecystectomy].

[Laparoscopic laser cholecystectomy]. In an ex vivo- and an animal study a Holmium-YAG laser, a Nd:YAG laser and a CO2 laser were examined. For laparoscopic cholecystectomy the pulsed Holmium:YAG laser proved to be the best instrument. Dissecting and haemostatic effects were connected when working with a power of 4 watts. Dissected area per minute was 112 mm2, gallbladder dissection (15 cm2) can be performed in 13 min. In the animal study the cholecystectomy lasted 30 min on average when the Holmium:YAG laser was used. Bleeding was not observed, development of smoke was tolerable.


[Therapy of splenic injuries by freezing and fibrin gluing. Animal experiment study]. The goal of this investigation was to improve the reliability of intraoperative and postoperative hemostasis and to observe the healing process after using a combined technique of tissue freezing followed by the application of collagen fleece and fibrin glue for the treatment of splenic ruptures. Grade II lesions were inflicted on the spleens of 15 swine. The bleeding wounds were frozen for 1 min at -60 degrees C using a cryosurgical device. Immediately afterwards the frozen lesions were covered with fibrin glue and collagen fleece and kept under slight compression. In every case complete hemostasis was achieved intraoperatively. The spleens of three animals each time were collected for gross and microscopic examination after 2 days and 1, 2, 5, and 6 weeks. A visceroperitoneal adhesion was observed in only one spleen, U-shaped viscerovisceral adhesions in five spleens. Superficial coagulation necroses could be detected microscopically only after 2 days and 1 week. Organization of the wounds, indicated by granulation tissue which contained siderophages, started in the 2nd week. There was distinct formation of collagen fibers after 5 and 6 weeks; only a residue of the collagen fleece was visible and the surplus fibrin glue was encapsulated. With this combined technique complete and safe hemostasis and a good subsequent healing process was achieved.


[Pneumococcus-induced septicemia in normal and splenectomized rabbits]. A new rabbit model to study the consequences of splenectomy in host resistance to induced pneumococcal septicaemia is presented. A simple, fast, non-invasive and complication-free technique of bacterial inoculation into the rabbit airways, using transcutaneous cannulation of the trachea, is also described. Sixteen normal and 24 splenectomised animals were used. The optimal dose of pneumonococcus (serotype B, type III) was found to be 1 x 10(7) organisms. The above dose of bacteria given transtracheally failed to cause signs of pneumonia or death in any of the normal rabbits. However the same dose introduced using the same technique resulted reproducibly in the death of the low-resistance splenectomised animals.


[Model of electromyographic study of small intestinal transplants in the rat]. Adequate motility of a small bowel transplant is a prerequisite for its resorptive function as well as its self-purging capacity. The literature contains some reports on changes in motility following denervation or transplantation, but none on the impact of acute rejection on motility of small bowel grafts. Therefore an experimental model was established to meet the following criteria: orthotopic position of graft, adequate nutritional status even when graft is functionally impaired, chronically implantable electrodes attached to graft and corresponding segment of native bowel, isogeneic or allogeneic set-up. 10 cm of proximal jejunum were transplanted from Lewis donors to five Lewis recipients in an orthotopic position just distal of the ligament of Treitz without resection of native small bowel. Three bipolar electrodes were sutured to the graft and the same number to the subsequent recipient bowel. Serial myoelectric measurements were taken until the end of week 3. From day 5 on, migratory myoelectric complexes independent of myoelectric activities of native bowel were recorded. Pacemaker frequency of the graft was found to be the same as that of the transsected native small bowel. This early reappearance of myoelectric activities makes this model suitable for comparative studies of small bowel transplant motility and in particular its changes during rejection, since even in strongly allogeneic combinations Lewis small bowel does not show histological signs of rejection before day 6.


[Proliferation kinetic studies in the syngeneic liver transplantation mode in the rat. Effect of re-arterialization on the transplant]. Successful orthotopic liver transplantation (OLT) can be achieved in the rat. We used bromo-2-deoxyuridine (BrdU) as a proliferation marker to document morphological differences between OLT with and without rearterialization. Animals with portal anastomosis alone had a significantly increased proliferation rate of hepatocytes, Kupffer cells, and bile duct epithelial cells, as indicated by strong staining with BrdU, 8 days post-transplant compared to animals with rearterialization. Regeneration of ischemically damaged liver parenchymal cells may account for this observation. Thus, OLT with rearterialization appears to be the more physiological transplant model.


[Doppler ultrasound measurement of blood circulation in the lower extremity of man. I. Normal values at rest and changes in trauma and vascular diseases]. A duplex ultrasonographic system was used to examine the common femoral artery in left and right legs of 49 fit young men and 15 fit young women, to find out the normal blood flow in the human leg during rest. By this method the diameter of the vessel and the time average velocity could be measured and the blood flow calculated. Each person underwent three examinations of each leg after 10-min rests. The average diameter of the common femoral artery was 91 +/- 10 mm (95 mm in the men and 80 mm in the women), the average blood flow 226.2 +/- 82.1 ml/min (235.9 ml/min in the men and 189.6 ml/min the the women). Compared with other methods used for femoral blood flow studies, we found the duplex ultrasound system reliable, non-invasive, and infinitely repeatable. This method therefore also seems appropriate for clinical studies. In a few tests carried out in trauma patients we found a pronounced increase in blood flow after a few days, and also an expected increase in a patient with arterio-venous fistula and an expected decrease in one with arterial occlusive disease. These examinations were performed to establish a method for further examinations of long-lasting blood flow alterations in trauma patients, which will be reported in another paper.


[Doppler ultrasound measurements of blood circulation of the lower extremity in man. II. Hemodynamics after trauma and operation]. A duplex ultrasound system was used to examine the blood flow of the common femoral artery in ten young patients with fractures of the lower extremities. Two patients had fractures of both legs. In eight patients the blood flow in the injured leg was comparable with that in the not injured leg. The flow alterations were measured on 3-7 different days over a period of 6-224 days after trauma. All patients showed a significant (to twice or three times that before injury) increase in blood flow in the injured leg a few days after trauma and/or operation, whereas the blood flow in the uninjured leg remained the same or even decreased a little. In the case of undisturbed fracture healing the change in blood flow disappeared within 8-12 weeks, corresponding to fracture consolidation. When fracture healing was delayed and/or further operations on the injured leg were necessary, the blood flow was increased for much longer. The hemodynamics of an injured extremity are compared with those reported elsewhere in the literature. The increased demand for oxygen or energy can explain the changes only in part. Another function of the reflective increase in blood flow could be the temperature increase in the extremity.


[Endosonography of stomach tumors].

[Endosonography of stomach tumors]. Based on own experience and on the published literature we report about indications and efficiency of endosonography (EUS) in gastric tumors. The following conclusions can be drawn at the present time. Submucous tumors can be clearly differentiated from extragastric compressions. Although the endosonographic aspect does not allow to formulate an etiologic diagnosis, EUS findings can give hints regarding the nature of the submucous tumor (e.g. leiomyoma, lipoma, cyst). In 75% of cases malignant submucous tumors can be visualized and a correct preoperative staging can be performed. EUS is of special importance in the description of gastric carcinoma. The pT stage can be correctly determined preoperatively in about 80% (69-92%) of cases. Accompanying inflammation in early gastric cancer can lead to overstaging. The sensitivity for local lymph node metastases reaches about 77% (50-88%). Gastric non-Hodgkin lymphomas can be excellently visualized with EUS. The sensitivity amounts to 90-100% and in about 90% of cases the extent of the tumor can be correctly determined preoperatively. The response to radio-chemotherapy of gastric non-Hodgkin lymphomas can be monitored easily with the method. At the present time EUS is the most sensitive imaging tool in visualizing and staging of gastric tumors. Its main advantage is the exact demonstration of intramural and paragastric alterations. However, despite the use of high ultrasonic frequencies and the excellent demonstration of even tiny details with EUS, biopsies for histologic evaluation are still mandatory, especially when dealing with gastric ulcer.


[Modified combined omeprazole/amoxicillin therapy for Helicobacter pylori eradication: a pilot study]. Patients with H.pylori positive peptic ulcer disease were treated with a two weeks regimen consisting of 20 mg omeprazole twice daily and 500 mg amoxicillin six times daily. Subsequently, an H2-receptor antagonist was started (300 mg ranitidine) at night time for four weeks. Before and one month after completion of antibiotic therapy an upper GI-tract endoscopy was performed for determination of H.pylori infection [biopsy urease test (BUT), specific culture and histologic demonstration]. A total of 12 patients completed the study protocol. H.pylori eradication, defined as a negative result in BUT, culture and histology) four weeks after completion of the combined omeprazole/amoxicillin treatment regimen was achieved in 91.6% (11 of 12 patients). Complete ulcer healing was confirmed in all patients. A stomatitis was observed in one female patient as a possible side effect of antibiotic treatment, but this did not necessitate discontinuation of therapy. Only complicated drug regimen with many side effects have been available so far for successful eradication of H.pylori. Thus, the present drug combination might prove as an effective therapeutic option in the future. These data, however, await confirmation in larger study population.


[Preoperative esophagogastroduodenoscopy before elective surgical therapy of symptomatic cholelithiasis]. We evaluated the data of 1143 patients who underwent preoperative gastroscopy or upper gastrointestinal series before elective surgical treatment of cholelithiasis between January 1, 1981 and December 31, 1990. On these 824 women and 319 men we performed 1064 (93.1%) gastroscopies and only 78 (6.8%) upper gastrointestinal series. The incidence of pathological findings was 30.2% (345 patients), with 68.3% findings of inflammatory nature. In 28 patients (2.5%) cholecystectomy or bile duct exploration was combined with an additional gastrointestinal surgical procedure. In 227 cases (19.8%) biliary surgery was followed by pharmacological treatment of the gastrointestinal disease. Because of the high incidence of simultaneous disease of the upper gastrointestinum we believe that routine preoperative gastroscopy is indicated before elective surgical treatment of cholelithiasis.


[Chilaiditi syndrome--general indications for surgical therapy?]. The usually asymptomatic Chilaiditi's syndrome is a rare anomaly which in most cases is detected accidentally by x-rays. We report two unusual cases with incarcerated interpositio coli hepato-diaphragmatica. Differential diagnosis and surgical management are discussed.


Bactericidal action at low doses of a new rifamycin derivative, 3'-hydroxy-5'-(4-isobutyl-1-piperazinyl) benzoxazinorifamycin (KRM-1648) on Mycobacterium leprae inoculated into footpads of nude mice. Among a series of newly-synthesized benzoxazinorifamycins, 2 of the 3'-hydroxy-5'-(4-alkyl-1-piperazinyl) derivatives, named KRM-1648 and KRM-2312, whose respective alkyl residues are isobutyl and isopropyl, were examined for efficacy against nude mouse-model leprosy. KRM-1648 completely inhibited the growth of leprosy bacilli inoculated into nude mouse footpads, even 6 months after the medication had been stopped, when given orally at a daily dose of 0.6 mg/kg, 5 or 6 times weekly, during 3-5 months postinoculation. In comparison, the growth inhibition by KRM-2312 was incomplete under the same conditions, though it was still stronger than that by rifampicin. Complete growth inhibition by KRM-1648 was also observed when it was given orally at a dose of 1 or 3 mg/kg twice weekly during the same period. In contrast, the growth inhibition by rifampicin was only slight at 1 mg/kg and partial at 3 mg/kg under the same condition.


Persistence of Mycobacterium leprae in the peripheral nerve as compared to the skin of multidrug-treated leprosy patients. Skin and nerve biopsies obtained from 18 multibacillary (MB) and 16 paucibacillary (PB) cases of leprosy who had been fully treated by the WHO regimen were assessed for bacterial load using different staining techniques. In addition skin and nerve homogenates of 10 MB cases were tested for 'persistor' Mycobacterium leprae using immunosuppressed mice. While significant amounts of integral bacilli and BCG cross-reactive antigen of M. leprae were detected both in skin and nerve tissues of all the MB cases (100%), 56% of skin and 62% of nerve biopsies of PB cases also showed the presence of BCG cross-reactive antigen. Detection of 'persistor' M. leprae in 2/10 skin biopsies (20%) and 3/10 nerve biopsies (30%) of MB cases was thought to be unexpectedly high after 2 years of MDT.


The international classification of impairments, disabilities and handicaps in leprosy-control projects. The use of a uniform language, which includes definitions of terms, is very important in the field of health care. It is important to have a common language for educational, research and communication purposes. Classifications can play a major role in the development of uniform reporting and registration systems. The purpose of this article is to familiarize leprosy workers with two classifications that are in common use in health care, a classification of diseases and a classification used to describe the overall health status of a person, and to relate the 3 terms that are used in the latter classification, impairments, disabilities and handicaps, to leprosy.


Risk factors among defaulters in the urban leprosy control centre of Thaketa Township in the city of Yangon, Myanmar, 1986. A total of 884 registered cases from the city of Yangon were retrospectively analysed. The defaulter proportion among cases registered for treatment at the Thaketa Health Centre was 34.16%. It was established that patient sex and occupation are not a factor in defaulting. Paucibacillary cases and cases with no disability are more likely to default.


Initial intensive therapy for multibacillary leprosy patients--in retrospect. We analysed the results of 4845 multibacillary (MB) patients being treated with multidrug treatment (MDT) in the Srikakulam District of Andhra Pradesh, India. Of these, 2309 (47.7%) patients were given an initial 14-day intensive therapy with rifampicin, clofazimine and dapsone, followed by the WHO recommended pulse therapy. The rest of the cases were given only pulse therapy. The improvement in terms of bacteriological clearance and the proportion of cases declared released from treatment (RFT) was found to be significantly higher among patients treated with only pulse therapy. Clinic attendance was found to be better and more regular in patients treated with intensive therapy, and no relapses were seen with either therapy. The implications of these findings on the operational aspects of programme implementation are discussed.


Immunotherapeutic potential of ICRC vaccine: a case control study. A bacteriological follow-up of 16 lepromatous patients with a high initial Bacteriological Index (BI) showed that in 8 randomly selected patients who received single doses of ICRC Vaccine (C44) at the onset of multidrug therapy, the average reduction of BI was from 4.4+ to 1+ in 2 years--3 of these patients became negative and 3 showed BI 1+ or less. Comparable bacteriological assessments in 8 non-vaccinated but otherwise similar patients showed an average reduction of BI from 4.7+ to 2.6+, i.e. consistent with the expected response to MDT in lepromatous patients. Here we discuss the role of immunotherapy and the selection of a desirable antileprosy vaccine in the context of fixed-duration MDT.


Healing rates of plantar ulcers in leprosy and diabetes. Comparison was made of wound healing time in a consecutive series of leprosy and diabetic patients with plantar ulceration. In the leprosy group, 66 of 70 (94%) ulcers healed in a mean time of 42.7 (+/- 36.1) days, and in the diabetic group, 75 of 80 (94%) ulcers healed in a mean time of 39.7 (+/- 32.1) days. Analysis of all healed ulcers using a general linear model found wound depth (p < 0.03), and wound diameter (p < 0.05) significantly related to ulcer healing time. Diagnosis, healing devices (cast, splint and cut-out sandal), age and sex were not significant. In diabetic subjects a regression model including depth, diameter and age explained 36% of the variation in healing time. A meaningful regression model was not found in leprosy patients.


Molecular mechanisms of attachment of Rhizobium bacteria to plant roots. Attachment of bacteria to plant cells is one of the earliest steps in many plant-bacterium interactions. This review covers the current knowledge on one of the best-studied examples of bacterium-plant attachment, namely the molecular mechanism by which Rhizobium bacteria adhere to plant roots. Despite differences in several studies with regard to growth conditions of bacteria and plants and to methods used for measuring attachment, an overall consensus can be drawn from the available data. Rhizobial attachment to plant root hairs appears to be a two-step process. A bacterial Ca(2+)-binding protein, designated as rhicadhesin, is involved in direct attachment of bacteria to the surface of the root hair cell. Besides this step, there is another step which results mainly in accumulation and anchoring of the bacteria to the surface of the root hair. This leads to so-called firm attachment. Depending on the growth conditions of the bacteria, the latter step is mediated by plant lectins and/or by bacterial appendages such as cellulose fibrils and fimbriae. The possible role of these adhesions in root nodule formation is discussed.


Mechanisms of the cytopathic action of actin-ADP-ribosylating toxins. Clostridium botulinum C2 toxin, Clostridium perfringens iota toxin, and Clostridium spiroforme toxin ADP-ribosylate actin monomers. Toxin-induced ADP-ribosylation disturbs the cellular equilibrium between monomeric and polymeric actin and traps monomeric actin in its unpolymerized form, thereby depolymerizing actin filaments and destroying the microfilament network. Furthermore, the toxins ADP-ribosylate gelsolin actin complexes. These modifications may contribute to the cytopathic action of the toxins.


cis-unsaturated fatty acids specifically inhibit a signal-transducing protein kinase required for initiation of sporulation in Bacillus subtilis. The initiation of sporulation in Bacillus subtilis is controlled by the Spo0A transcription factor which is activated by phosphorylation through a phosphorelay mechanism that is dependent upon the activity of one or more protein kinases. The enzymatic activity of one of these protein kinases, KinA, was found to be inhibited in vitro by certain fatty acids. The most potent inhibitors have at least one unsaturated double bond in the cis configuration and a chain length of 16-20 carbon atoms. Homologous isomers with a trans double bond are not inhibitory. Saturated straight- or branched-chain fatty acids are either much weaker inhibitors or have no effect. The inhibitors prevent autophosphorylation of KinA and are non-competitive with ATP. B. subtilis phospholipids were found to contain at least one as yet unidentified type of fatty acid that, when present in an unesterified form, inhibited KinA. The results suggest that the concentration of a specific unsaturated fatty acid may act as a signal linking the initiation of sporulation to the status of membrane synthesis and septation or some other specific membrane-associated activity.


Cloning and characterization of a gene encoding flagellin of Listeria monocytogenes. The gene, flaA, encoding the flagellin protein of Listeria monocytogenes (strain 12067) has been isolated from an expression library in Escherichia coli using a flagellin-specific monoclonal antibody. DNA sequence analysis of a positive clone revealed the presence of an open reading frame of 287 amino acid residues with a calculated molecular mass of 30.4 kDa. Comparison of this sequence with flagellins from other bacteria showed a significant degree of homology in both the N- and C-terminal parts of the protein. The flagellin mRNA was determined to be 1 kb in size, which is the expected size for a monocistronic mRNA, and the temperature-dependent expression of flagellin was found to be regulated at the transcriptional level. Southern blot analysis, using the flagellin gene as probe, indicated that L. monocytogenes can be divided into two groups. These groups correspond to the flagellar antigens AB and ABC, respectively, as well as to the two types of L. monocytogenes based on the DNA sequence of the listeriolysin gene.


An inverted repeat preceding the Bacillus subtilis glpD gene is a conditional terminator of transcription. The Bacillus subtilis glpD gene, encoding glycerol-3-phosphate (G3P) dehydrogenase, is preceded by a promoter and an inverted repeat which is located between the promoter and the glpD coding region. The inverted repeat acts as a transcriptional terminator in vitro. Expression of glpD is induced by G3P in the presence of the glpP gene product. Full-length glpD transcripts can be detected only in glycerol-induced cells. The major glpD transcript is initiated from the glpD promoter but minor amounts of larger transcripts, possibly initiated at upstream glp promoters, can also be found. In uninduced cells short transcripts are present, corresponding to initiation at the glpD promoter and termination at the inverted repeat. Upon induction, these short transcripts disappear and are replaced by full-length glpD transcripts. The 3'-ends of full-length glpD transcripts were mapped to an inverted repeat located immediately downstream of glpD. These results show that glpD of B. subtilis is regulated by termination/antitermination of transcription.


Gene disruption identifies a 290 kDa cell-surface polypeptide conferring hydrophobicity and coaggregation properties in Streptococcus gordonii. The C-terminal coding region of the gene (denoted cshA) encoding a high-molecular-mass (290 kDa) cell-surface polypeptide in the oral bacterium Streptococcus gordonii was cloned and sequenced. Insertion of ermAM into the S. gordonii chromosome at the 3' end of the coding region of cshA led to the production of isogenic mutants that secreted a truncated form (260 kDa) of the CshA polypeptide into the growth medium. Mutants had reduced cell-surface hydrophobicity and were impaired in their ability to coaggregate with oral actinomyces. The results identify a carboxyl terminus-anchored cell-surface protein determinant of hydrophobicity and coaggregation in S. gordonii.


The TraM protein of the conjugative plasmid F binds to the origin of transfer of the F and ColE1 plasmids. The gene encoding the TraM protein of the conjugative plasmid F was cloned, overexpressed and the gene product was purified. The TraM protein was found in the cytoplasm of cells carrying the F plasmid with a smaller amount in the inner membrane. DNase I footprinting experiments showed that the purified protein protects three regions in the F oriT locus with different affinity for the upper and lower strands of DNA. A 15-nucleotide motif was identified within the protected regions that represented the DNA-binding site. The TraM protein was also found to bind to a sequence in the oriT region of the non-conjugative plasmid ColE1 that resembles the three binding sites in the F oriT region.


Sequence alterations affecting F plasmid transfer gene expression: a conjugation system dependent on transcription by the RNA polymerase of phage T7. We constructed derivatives of the Escherichia coli conjugative plasmid F that carry altered sequences in place of the major transfer operon promoter, PY. Replacement of PY with a promoter-deficient sequence resulted in a transfer-deficient, F-pilus-specific phage-resistant plasmid (pOX38-tra701) that could still express TraJ and TraT; TraY, F-pilin, TraD, and TraI were not detectable on Western blots. On a second plasmid (pOX38-tra715) we replaced PY with a phage T7 late promoter sequence. In hosts carrying a lacUV5-promoter-regulated T7 RNA polymerase gene, all transfer-associated properties of pOX38-tra715 could be regulated with IPTG. After induction, pOX38-tra715 transferred at the wild-type frequency, expressed normal numbers of F-pili and conferred sensitivity to pilus-specific phages. No adverse effects on cell viability were apparent, and additional mutations could easily be crossed onto pOX38-tra715. A traJ deletion (pOX38-tra716) had no effect on the IPTG-induced transfer phenotype. Insertion of cam into trbC, resulted in a mutant (pOX38-tra715trbC33) which, after induction, exhibited the same phenotype associated with other trbC mutants; it could also be complemented by expression of trbC in trans. With pOX38-tra715 or its derivatives, we were able to label specifically the products of tra genes located throughout the long tra operon, by using rifampicin. This feature can be used to investigate transfer protein interactions and to follow changes in these proteins that are associated with conjugal mating events.


Determination of a specific region of the purine-cytosine permease involved in the recognition of its substrates. Three u.v.-induced mutants of the purine-cytosine permease gene of Saccharomyces cerevisiae, with altered apparent Michaelis constant of transport (Kmapp), were cloned and sequenced. One of the mutants had extensive nucleotide replacement, whereas the other two had a single mutation. To evaluate the contribution of the different amino acid replacements to the phenotype of the complex mutant, simpler mutants were created by site-directed mutagenesis. All the amino acid replacements found in the segment from amino acids 371 to 377 inclusive, contribute to the determination of the phenotype. According to the model postulated this segment lies on the cell surface. In particular, amino acids at position 374 and 377 modulate the affinity of the permease towards its substrates. In the wild-type, when asparagine is present at both of these positions, the lowest Kmapp values are found.


Determinants of an unusually stable mRNA in the bacterium Myxococcus xanthus. Myxococcus xanthus is a Gram-negative bacterium which has a complex life cycle that includes development (fruiting body formation). The gene for myxobacterial haemagglutinin, mbhA, is developmentally regulated and highly expressed. In this report we show that the mbhA mRNA is exceptionally stable for a prokaryotic organism, exhibiting a chemical half life (t1/2) of 150 min at 18 h of development. The mbhA mRNA was not stable in vegetatively growing cells nor was it stable when expressed in Escherichia coli. We have used site-directed mutagenesis of the mbhA gene to analyse some of the determinants which mediate the stability of the mbhA transcript. Sequences within the 3'-untranslated region (3'-UTR) were found to be crucial for mRNA stability. This region of mRNA can potentially form an extremely stable stem-loop structure immediately adjacent to the translational stop codon. A deletion within this region caused a 10-fold increase in the decay rate of the transcript. Furthermore, conditions which were associated with reduced mbhA translation or mutations that caused premature termination of translation drastically reduced mRNA stability even in the presence of the wild type 3'-UTR. These results suggest that a significant aspect of mbhA mRNA stability involves a synergistic interaction of the translational machinery with sequence elements within the 3'-UTR.