Management of bony defects in open grade III fractures.

Early soft tissue reconstruction is recommended in treating open grade III fractures. Yet, there are still controversies regarding the way to handle bony defects when the soft tissue reconstruction is completed. This is a retrospective study to evaluate various methods of treating bony defects after soft tissue reconstruction in open fractures. From May 1985 to June 1988, we experienced 16 cases of open grade III fractures of the extremities with bony defects which received either local treatment with flap transfers (five cases) or free flap transfers (11 cases). The bony defects ranged from 3 cm to 10 cm in length. Cancellous bone grafting was performed in nine cases, and two of them received internal fixation at the same time. Seven cases received vascularized bone grafts (six fibulas and one iliac bone). Our results showed that bone grafting, with either a conventional cancellous bone graft or a vascularized bone graft, is useful in treating bony defects after soft tissue reconstruction. Internal fixation of the fractures after removing an extraskeletal fixator has the disadvantages of soft tissue breakdown and of possible infection. Early posterolateral bone grafting was beneficial if a cancellous bone graft was used on the tibia. Double free tissue transfers were quite useful, and we did not lose any free tissue transfers in our series. Vascularized bone grafts should be considered in cases where the osseous gap is greater than 6 cm, or where the recipient bed is bad.


Clinical efficacy of transluminal extraction coronary atherectomy.

Mechanical revascularization has revolutionized the treatment of coronary artery disease. The transluminal extraction-endarterectomy catheter (TEC) system was developed recently and permits the excision and extraction of atherosclerotic lesions of the coronary artery and the bypass graft. The system includes a motor-driven flexible torque catheter with rotating conical shaped stainless steel blades distally. The proximal end of the TEC system consists of a mechanical housing which controls the vacuum, the rotating cutter (750 RPM) and the cutter excursion (4 cm). After safety testing of the TEC system in dogs and human peripheral arterial disease, percutaneous transluminal coronary atherectomies employing the TEC have been performed in 25 patients (six women, 19 men, mean age, 64 +/- 12 years) with the diameter of the stenosis greater than or equal to 75% in one or more coronary arteries. Twenty-eight atherosclerotic lesions were treated in 26 native coronary arteries. The overall success rate (less than 50% residual stenosis in all lesions) was 92%. There was one instance of hematoma proximal to the excised target lesion, and one instance of atherectomized debris embolization. There were no instances of dissection, perforation, coronary spasms or death related to the procedure. The results indicate that the TEC system can be used safely and effectively to treat coronary artery disease with a high success rate.


Lumbar lordosis: normal adults.

Lumbar curvatures in 149 normal adults from the general population were studied. There were 76 men and 73 women with an average age of 50 years. The mean values of lumbar lordotic angle (LLA), lumbosacral angle (LSA) and sacral inclination angle (SIA) were 33.2 +/- 12.1 degrees, 11.4 +/- 4.7 degrees and 26.4 +/- 10 degrees, respectively. A high correlation was noted between LLA and SIA (r = 0.883, p = 0.0001). LLA is an ideal parameter for the evaluation of lumbar lordosis. The normal value of LLA can be defined as 20-45 degrees with a range of 1 SD. No significant differences were noted in these three angles between males and females in any age group (LLA, p = 0.647; LSA, p = 0.80; SLA, p = 0.189). Also, X-ray findings indicated there were no significant differences between these three angles in spondylotic spines and those spines with a normal appearance from X-ray finding. The average LLA increased with age. Significant lumbar lordotic angle differences were noted between those patients less than 35 years of age and those greater than 60 years, as well as in the 35-60 age group and the greater than 60 age group (p = 0.0056).


Locomotor strategies before independent walking: prospective study of 50 mentally retarded children.

To investigate the association between pre-walking locomotor strategies and psychomotor developments in children with mental retardation (MR), 50 children with non-specific MR were included in this study. There were 29 boys and 21 girls, 96% of whom had moderate to severe MR. They were followed from 4-53 months to 25-99 months of age, and their follow-up periods ranged from 10 to 48 months (mean 30 months). According to the pre-walking locomotor strategies, these children were categorized into three groups: the crawling group (n = 34) who used crawling or creeping as their main locomotion pattern before independent walking; the shuffling group (n = 9) who used shuffling prior to independent walking; and the direct-walking group (n = 7) who did not have any other locomotor strategies except rolling. In almost all motor developmental milestones, children in the direct-walking group developed earlier than those in the crawling and shuffling groups. Children in the crawling group had more advanced developments than those in the shuffling group. The difference in the mean ratio developmental quotients of the Bayley Mental Scale among the three groups was not significant. The present study showed that crawling may not be a necessary prerequisite for early ambulation or better cognitive function in MR children.


Pelvic actinomycosis with colo-ileo-vesical fistula formation: report of a case.

Pelvic actinomycosis with multiple fistular formation is rarely reported in the literature. We herein present a case of pelvic actinomycosis with sigmoid colo-ileovesical fistulae in a 36-year-old intrauterine device (IUD) user. She was admitted to the hospital because of general malaise, weight loss and bilateral palpable adnexal masses. Sonography showed bilateral adnexal masses which contained many echolucent spots. A barium enema examination revealed sigmoid colo-ileo-vesical fistulae. A computed tomographic scan showed bilateral cystic adnexal masses, bilateral hydronephrosis and hydroureter. Preoperatively, pelvic malignancy was suspected. An exploratory laparotomy was performed. Bilateral tubo-ovarian abscesses with extensive adhesions were found. Pathologic examination of the operative specimen revealed pelvic actinomycosis. The patient was treated with penicillin for 14 weeks and had a stable clinical course.


CT findings of pediatric thoracic actinomycosis: report of four cases.

Thoracic actinomycosis is an uncommon disease, which may mimic malignancy, lymphoma or tuberculosis of the chest. In the past three years, four cases of thoracic actinomycosis have been found in children at our hospital. Their computed tomography (CT) findings included pulmonary infiltrates, a chest wall mass, pleural and pericardial effusion, mediastinum involvement and rib changes. Although the final diagnosis of Actinomyces infection depends on a bacterial culture and pathology, CT can play an important role in establishing the diagnosis and evaluating the extent of the disease.


Acute pseudo-obstruction of the colon following partial splenic artery embolization: report of a case.

This is a case of a 45-year-old woman suffering from liver cirrhosis with hypersplenism who received a partial splenic artery embolization with gelfoam pieces. Thrombocytopenia and leukopenia improved immediately after treatment. Marked abdominal distention, metallic bowel sounds and combined small and large bowel distension with air-fluid levels as shown on a plain abdominal X ray were noted on the fifth day after the embolization procedure. Laparotomy was performed. No mechanical obstruction was found during surgery. Acute pseudo-obstruction of the colon (Ogilvie's syndrome) was diagnosed. This condition was considered to be a complication of the partial splenic artery embolization and has not hitherto been reported.


Circumaortic left renal vein: report of a case.

Circumaortic left renal vein is an uncommon anomaly which has not been previously reported in Taiwan. We describe a case of circumaortic left renal vein in a 15-year-old girl who presented with intermittent hematuria. Angiogram revealed double left renal veins in the form of a venous collar. Computed tomography depicted the preaortic and retroaortic left renal veins clearly. In a survey of hematuria, circumaortic renal vein should be taken into consideration in a differential diagnosis, and an appreciation of the specific radiologic findings can help to establish the diagnosis easily. In addition, preoperative recognition of this uncommon anomaly is of supreme importance in a retroperitoneal operation in order to avoid massive bleeding.


Postnatal development of GABAergic neurons in the gerbil cochlear nucleus: pre-embedding and post-embedding immunocytochemical staining.

The purpose of this investigation was to compare the results obtained when plastic-embedded sections and vibratome-sliced sections were used to localize gamma-aminobutyric acid immunoreactivity (GABA-IR) in the gerbil cochlear nucleus (CN) during postnatal development. GABA-IR was mainly located in the perikarya of the neurons of the gerbil CN. At two days old, GABA-IR was found in plastic sections. No discernible GABA-IR was found in the vibratome slices at three to four days. At eight days, anti-GABA labeled cells were randomly located in the superficial and deep layer of the dorsal cochlear nucleus (DCN) in the vibratome sections, whereas they were mainly accumulated in the vicinity of the granule layer between the junction of the DCN and posteroventral cochlear nucleus (PVCN) in the plastic sections. At 14 days, anti-GABA labeled cells were reduced in number in the superficial third of the DCN in the vibratome slices and reduced in number in the junction between the DCN and PVCN in the plastic sections. At that time, a striking change was the formation of lamellation of the anti-GABA labeled cells mainly located in the middle third (fusiform layer) of the DCN in the plastic sections. A similar pattern of lamellation was found in the vibratome slices at 17-18 days. At three, four, six and eight weeks, the anti-GABA labeled cells were mainly located in the ventral part of the DCN close to the choroidal plexuses and the ventral part of the PVCN in the plastic sections. At six to 15 months old, distinct anti-GABA labeled cells were located in the fusiform layer of the DCN and scattered in the PVCN in the plastic sections. Another striking change at this age was that numerous vacuoles with a mesh-like network were present in the PVCN.


Plasma renin activity, aldosterone level, serum and urinary electrolytes in normal pregnant women aged 35 and older.

Plasma renin activity (PRA), the plasma aldosterone (PA) level, and serum and urinary electrolytes were measured in 39 elderly pregnant women of greater than or equal to 35 (Group 1) and in 60 pregnant women less than 35 (Group 2) every four weeks from the 20th week of gestation to the fourth week postpartum. The PRA and PA levels increased in both groups. The PA levels increased after the 20th week and reached a peak at the 32nd week of gestation, while PRA decreased after the 20th week of gestation. This dissociation was observed in both groups. Daily urinary sodium excretion in Group 1 was higher than that of Group 2, while daily potassium excretion was not different between the two groups. Higher aldosterone secretion was observed after the 20th week of pregnancy in Group 1. It is concluded that pregnancy in older women is associated with higher sodium excretion and aldosterone secretion.


Peripheral and intracardiac concentrations of atrial natriuretic peptide in patients with heart disease.

To study the relationship between the plasma concentration of the atrial natriuretic peptide (ANP) and heart function and to discern the secretion pathway of ANP, we determined the peripheral plasma concentration of ANP in 18 heart patients and the intracardiac concentration of ANP in six heart patients during cardiac catheterization. All plasma was extracted through a Sep-Pak C18 cartridge by acid alcohol. The concentration of ANP was determined by a sensitive and specific radioimmunoassay method. We found that the plasma ANP concentration (pg/mL) in heart patients (ranging from 12 to 139, mean = 36.1 +/- 28.9) was statistically higher than that in normal adults (ranging from 8 to 20, mean = 12.4 +/- 3.3, n = 16; p less than 0.05). The ANP level in heart patients was inversely correlated with the left ventricular ejection fraction (r = -0.53, p less than 0.05) evaluated by radionuclide angiocardiography. The intracardiac level of ANP was 88 +/- 58 at the superior vena cava, 77 +/- 55 at the inferior vena cava, 124 +/- 68 at the right atrium, 98 +/- 64 at the right ventricle, 107 +/- 58 at the pulmonary artery, 98 +/- 52 at the left ventricle, 109 +/- 73 at the aorta and greater than 351 at the coronary sinus. In conclusion, ANP is mainly secreted via the coronary sinus into the atrial cavity. The peripheral plasma ANP concentration is higher in heart patients. It increases as the left ventricular ejection fraction decreases. Therefore, plasma ANP concentration may be a useful indicator for the assessment of cardiac function.


Metformin-induced lactic acidosis: report of a case.

Lactic acidosis associated with diabetic patients receiving metformin therapy is rare but may cause significant morbidity and mortality. In nearly all reported cases of metformin-associated lactic acidosis, contraindications to its use were noted, especially renal insufficiency. We describe a 59-year-old diabetic man treated with metformin for more than three years. During the third year of use, he experienced progressive renal function impairment, and during the final month of use, he became azotemic. He was maintained on continuous ambulatory peritoneal dialysis. Several days prior to admission, he suffered from epigastralgia, nausea and vomiting, followed by progressive dyspnea which was Kussmaul in nature. Profound hypotension developed and he sank progressively into a coma. Wide-anion gap metabolic acidosis without ketonemia was detected. His blood lactate level was elevated and metformin-induced lactic acidosis was substantiated. An elevated plasma metformin level of greater than 50 mg/mL was determined later by high-performance chromatography. Rigorous treatment including bicarbonate therapy, bicarbonate hemodialysis and vasoactive agents as well as supportive measures were provided. With a return of pH to normal, the hypotension resolved and his consciousness level slowly improved. Our patient survived this disastrous event, but some neurologic sequelae remained. In order to avoid this life-threatening metabolic disturbance, patients with any contraindications should not be prescribed metformin.


[Investigation of the profile of selected trace metals in genetically obese (ob/ob) and lean (+/?) mice].

Trace metals are known to have important effects on the activity of metalloenzymes and insulin secretion and to be involved in the etiology of various diseases. This study was designed to investigate the distribution and concentration of selected trace metals in the tissues of genetically obese mice, which were known to have hyperinsulinemia. Different phenotypes (ob/ob; +/?) and sexes of 4, 8, 12 week-old mice were killed by decapitation. Metal levels (Zn, Cu, Cd, Cr) in the brain, liver, serum, hair and carcass were determined by an atomic absorption spectrometer. The results showed that obese mice had lower concentrations of zinc in the serum, hair, liver and carcass than lean controls (p less than 0.05), but there was no difference in the brain. Obese mice also had a low carcass cadmium concentration (p less than 0.01), which depended on the sex-age interaction. When expressed in terms of total content, obese mice had higher total liver zinc and carcass chromium contents (p less than 0.05). Obese mice had a higher total carcass copper content at 8 weeks of age. Serum and carcass zinc were showed to be inversely related to body fat in obese mice. The results indicate that zinc may play a special role in thermoregulation and fat metabolism in the liver of obese mice. The tissue distribution and absorption of zinc may have an important correlation in the development of obesity. The roles of copper, cadmium and chromium are still obscure, the related regulations are still open for further questions.(ABSTRACT TRUNCATED AT 250 WORDS)


[An animal model for colon cancer metastases to the lung and establishment of metastatic cancer cell line].

Death, in most colon cancer patients, is not caused by their primary tumors. Instead, metastasis is the major cause of morbidity and death. There are few appropriate animal models to investigate the complexity of the metastatic process. Therefore, the pathogenesis of the metastasis process is not clearly understood. We injected SW480 cells into the cecal wall of athymic nude mice to develop an animal model for colon cancer metastasis and to produce a metastasizing tumor. After metastatic foci formed in the murine lung, we established a metastatic cancer cell line, named CC-ML1, by in vitro primary culture. The characteristics of CC-ML1 were (1) a shorter doubling time; (2) a slightly higher metastatic rate and number of colonies; and (3) less morphological variation than those of SW480. Our research suggests that a higher metastatic potential may be found in CC-ML1 than in SW480 after the advanced cycles of the metastatic process. Thus, the primary animal model in this study may be useful in future studies of cells with a high metastatic potential in the pathogenesis of colon cancer metastases.


[A study of perigastric lymph node metastasis in patients with early gastric cancer].

The prognosis after a gastrectomy for early gastric cancer (EGC) is favorable. Among 115 patients with early gastric cancer since 1983, 96 patients have received a radical gastrecomy (RG) and 19 have received a simple gastrectomy (SG). The cancer was confined to the mucosa in 47 cases and had invaded into the submucosa in 68 cases. Perigastric lymph nodes metastasis was observed in 17 patients (14.8%). The metastatic rate for mucosal cancer was 2.1% and that of submucosal cancer was 23.5% (p less than 0.005). Advanced nodal metastasis was occasionally visible in a few patients with submucosal cancer. The macroscopic pattern, size, primary site and differentiation of the cancer and the age and sex of the patients did not influence the rate of nodal metastasis. A follow-up study of 61 patients with EGC operated on before 1988 was carried out, the 5-year survival rate of RG was 92.9% and that of SG was 84.2% (p = 0.3109). The 5-year survival rate for mucosal cancer for RG was 100% and that for was 90.9% (p = 0.3048). The 5-year survival rate for submucosa] cancer for RG was 89.3% and that for SG was 75.0% (p = 0.0685). We suggest that RG be mandatory for EGC unless the depth of invasion is confined to the mucosa in the preoperative diagnosis, or if gross advanced metastasis is present to obtain more accurate postoperative staging and to achieve a curative resection. However, SG is still an alternative treatment for EGC if a patient's condition is poor or for extremely old patients.


[Intraoperative pleural lavage in lung cancer patients].

In lung cancer patients, pleural effusion may or may not be present even if cancer cells have invaded the pleura. Using the traditional staging methods, not all pleural metastasis can be detected before the presence of pleural effusion. We performed intraoperative pleural lavage on 38 patients who underwent a thoracotomy due to tumors of the lungs. The first lavage (lavage 1) was performed just after the pleural cavity was open; the second lavage (lavage 2) was performed after all surgical procedures were completed (including lung resection and mediastinal lymph node dissection). Totally 12 of the 32 primary lung cancer patients were found to have positive lavage 1 cytology; 13 had positive lavage 2 cytology. In 13 patients whose visceral pleura was invaded by tumor, 11 had a positive lavage 1, 10 had a positive lavage 2. For prediction of positive lavage 1 cytology from visceral pleural involvement, the accuracy was 90.6%, the sensitivity was 84.6%, the specificity was 94.7%, the positive predictive rate was 91.7%, the negative predictive rate was 90.0%. For lavage 2, the accuracy was 81.2%, the sensitivity was 76.9%, the specificity was 84.2%, the positive predictive rate was 76.9%, the negative predictive rate was 84.2%. Stage of disease and cell type seemed to have no relationship to the incidence of positive lavage fluid cytology. Further investigation is recommended to evaluate the prognosis of patients with positive lavage fluid cytology.


[Secular trends in mortality for cerebrovascular diseases in Taiwan (1959-1989)].

Cerebrovascular disease (CVD) is predominantly a disease of the elderly, and its morbidity effects increase with advancing age. In Taiwan, the increasing proportion of the elderly, as a result of medical progress and improved health care in the past 30 years, is largely responsible for the apparent increase in the number of CVD deaths. From 1963 to 1981, CVD was the leading cause of death. The crude mortality rate (CMR) and age-specific mortality rate (ASMR) of CVD by sex were derived from vital statistical data from 1959 to 1989 in Taiwan. The age-adjusted mortality rate (AAMR) using the standard world population of WHO and the cumulative mortality rate (CUMR) from birth to less than 80 years of age were calculated. Before 1983, the total number of CVD deaths had increased steadily for 30 years. In 1989, the CMR was 76.6/100,000 in men and 67.7/100,000 in women. The highest AAMR was 158.5/100,000 in 1973 for men and 130.2/100,000 in 1972 for women, and the lowest AAMR was 91.3/100,000 in 1989 for men and 81.1/100,000 in 1972 for women. The highest CUMR was 26.3% in 1968 for men and 20.8% in 1972 for women, and the lowest CUMR was 14.5% in 1989 for men and 13.6% in 1989 for women. The AAMR and CUMR for both sexes reached a maximum in 1972 and began to decline thereafter. The declines in AAMR and CUMR were averaging 2%/yr for both sexes after 1972 and were averaging 5%/yr for men and 4%/yr for women after 1983. This declining trend in CVD deaths in Taiwan began later and has been slower than similar trends in Japan and the U.S.


[Effects of PGE1 on penile blood flow].

From February to July in 1989, 47 patients came to our O.P.D. with the chief complaint of impotence. The average age was 48.3 +/- 10.7 y/o. We applied intracoporeal injection of prostaglandin PGE1 (20 mg); and evaluated its penile blood flow effect by color duplex scanning (Acuson 128). The erectile responses of the test showed that: 6 patients (12.8%) had normal response: and 16 patients (34%) had imperfect response. Altogether, the total positive response rate was 46.8%, and 25 patients (53.2%) showed impaired response. The onset of response was 9.1 +/- 3.6 minutes and the duration of erection was 59.2 +/- 24.7 minutes. The percentage of diameter change of both deep arteries after injection was Rt: 58.1 +/- 41.5%; left: 52.3 +/- 35.6%. The peak velocity of right cavernosal arterial flow after intracoporeal injection was 35.5 +/- 15.9 cm/sec; and that of the left side was 33.2 +/- 16.9 cm/sec. There was no correlation between the increment of peak velocity of the deep arterial flow and the erection grade. The same phenomenon was also found between the increased change in the diameter of the deep artery and the erection grade. 16 patients (34.1%) experienced tolerable pain during the procedure. Two patients (4.3%) experienced dizzines and discomfort due to venous leakage. No priapism was found. This study suggests PGE1 may be an excellent potential alternative to other vasoactive drug with less complication in the diagnosis and treatment of impotence. But the cost and stability were its shortcoming.(ABSTRACT TRUNCATED AT 250 WORDS)


[Thyroid function tests in acute drug intoxication].

It is well known that thyroid function tests may be changed in non-thyroidal illnesses. To understand the influence of acute drug intoxication on thyroid function tests, 31 drug intoxicated patients without previous thyroid disorders and systemic diseases were included in our study. T3, T4, TSH, and resin T3 uptake were checked as soon as they arrived at our emergency service and were compared to that of 58 healthy volunteers. Within 31 patients, 14 were intoxicated by organophosphorous compounds, 6 by sedatives and hypnotics, 3 by strong acid, 2 by paraquet, 2 by rodenticides (warfarin), 2 by lysol and the other 2 were intoxicated by acetaminophen. The mean T3 and TSH levels were significantly lower in the drug intoxicated group. Among the 31 patients, 14 (45.2%) had a low T3, 2 (6.5%) had a low T3 and T4, and 6 (19.3%) had an elevated T4. All of the patients with an elevated T4 were intoxicated by organophosphates. If we divided the 31 patients into 2 subgroups: organophosphate intoxicated group and non-organophosphate intoxicated group, T4 and FT4I were significantly higher in the former group. Thyroid function tests became normal after treatment in 27 patients, discharged in good general condition. T3 and T4 became extremely low in 4 patients before they expired. The present study confirms that acute drug intoxication, like other non-thyroidal illnesses, affects thyroid function tests. Acute organophosphate intoxication may cause transient hyperthyroxinemia.


[Uterine lipoleiomyoma with calcification: report of a case].

Uterine lipoleiomyoma is uncommon and has received little attention from gynecologists. We report a case of uterine lipoleiomyoma with subsequent pelvic abscess after rupture of the appendix. Its clinical picture mimicked uterine malignancy. The patient was a 60-year-old woman, who presented with a 2-week period of lower abdominal pain which was exacerbated 10 days later. After admission, gynecologic examination revealed a large pelvic mass of firm consistency. On a plain film of the abdomen, there was a large calcified mass in the pelvis. Pelvic ultrasound demonstrated a 15 yen 12 yen 12 cm mass with strong echogenicity in the margin, but the central component of the mass was attenuating and revealed a poorly defined echogenic mass. A computed tomography scan of the pelvis demonstrated a well-encapsulated mass that was predominantly the density of fat (-65 Hounsfield unit) in the central part, with calcification present in the peripheral layer of the mass. There was also a cystic lesion measuring 3 yen 2.5 yen 1.5 cm in the right adnexal area. A preoperative diagnosis of uterine lipoleiomyoma with necrosis, liposarcoma of the uterus or teratocarcinoma of the ovary was made by the CT scan. A diagnostic dilatation and curettage (D&C) revealed a uterine cavity length of 12 cm by sounding, and the specimen showed no malignant tissue. Therefore, the preoperative suspicion of ovarian teratocarcinoma could be excluded. The patient underwent an exploratory laparotomy, multiple pockets of pelvic abscess, enlargement of the uterus and induration of omentum surrounding the appendix were found.(ABSTRACT TRUNCATED AT 250 WORDS)


[Chronic chromate intoxication with renal tubular damage--report of a case].

A 46-year-old male chromium plating worker visited our hospital due to rhinorrhea, sneezing and cough with blood-tinged sputum for more than 10 years. He also had skin ulceration and chronic dermatitis on both hands Medical therapy was inefficient. Physical examinations revealed nasal septum perforation, severe inflammation of the nasopharynx cavity, and eczema of both hands. Laboratory investigations showed significant tubule proteinuria, enzymuria, hypercalciuria, etc. It is evident that renal tube damage was present in this patient. The blood chromium level was 25 ng/mL, and the 24-hour urine chromium excretion level was 2.8 mg/day. A pulmonary function test showed reduced functional residual capacity (FRC), which may be due to either long-term smoking or chromate acid exposure. To our knowledge this is the first case of renal tubal damage induced by chronic chromate intoxication Taiwan. Further evaluation of the occupational safety and health of chromium plating workers is needed on this island.


Ubiquitin-reactive axons have a widespread distribution and are unrelated to prion protein plaques in Creutzfeldt-Jakob disease.

The amyloid plaques of Alzheimer disease (AD) are surrounded by dystrophic axons that contain ubiquitinated dense bodies. To investigate whether deposits of other types of amyloid cause axonal degeneration we studied 5 cases of Creutzfeldt-Jakob disease (CJD) with immunocytochemical methods using ubiquitin and prion protein (PrP) antisera. One of these cases contained PrP plaques in the cerebellum. In all cases dystrophic axons, which contain ubiquitinated dense bodies, were observed in neocortical and cerebellar grey matter, in absence of PrP-reactive amyloid deposits. Only a minority of PrP plaques present in the cerebellum was associated with ubiquitin positive neurites. The results indicate that, unlike in AD, the occurrence of ubiquitinated dystrophic axons is independent from amyloid deposition in CJD and is likely to be a primary phenomenon.


Dissociated neurons of the pupal honeybee brain in cell culture.

Primary cell cultures were prepared from specific regions of the pupal honeybee brain which are involved in proboscis extension learning. Defined areas could be dissociated purely by mechanical treatment. We show that cultured neurons regenerate new neurites and remain viable for up to three weeks in a serum-free, chemically-defined medium. Several labelling techniques were employed to identify subpopulations of cultured neurons. For example, acetylcholinesterase staining; fluorescent beads to distinguish identified cell populations of co-cultured brain areas; various markers for surface antigens such as a monoclonal antibody to olfactory projection neurons of the antennoglomerular tracts and monopolar cells of the optic lobes, as well as anti-HRP immunoreactivity and alpha-bungarotoxin binding; and various antisera for detecting transmitter phenotype. The appearance of transmitter-immunoreactive cells agreed closely with that expected from their known distribution in situ. Our results suggest that cultured cells retain surface properties and transmitter phenotype of their in vivo counterparts, despite differences in basic morphology. Thus our culture system provides the important initial step for future in vitro investigations of the cellular and electrophysiological properties of neurons mediating proboscis extension learning.


The effect of 5-lipoxygenase inhibition on blood-brain barrier permeability in experimental brain tumors.

To determine if leukotrienes are important mediators of vascular permeability in brain tumors, the effect of 5-lipoxygenase inhibitors on blood-tumor barrier permeability in rats harboring HK Walker 256 brain tumors was examined using quantitative autoradiography with alpha-14C-aminoisobutyric acid. The 5-lipoxygenase enzyme converts arachidonic acid to leukotrienes. Three 5-lipoxygenase inhibitors were utilized: BW755C, nordihydroguaiaretic acid, and AA-861. All three 5-lipoxygenase inhibitors significantly decreased vascular permeability both within the tumors and in brain adjacent to tumor. This suggests that capillary permeability in and adjacent to tumors is influenced by endogenous leukotrienes and that leukotrienes play an important role in brain tumor edema.


Oral benzodiazepines and conscious sedation: a review.

A large number of benzodiazepines have been studied for use as sedatives and for their anxiolytic potential as premedicants for outpatient surgery. Potent, new, orally-administered drugs with short half-lives, rapid onset, and minimal residual effects have been developed. Dose-dependent amnesia is also produced by some of these agents. Advances in understanding receptor physiology have shed light on specific pharmacologic activities and aided the discovery of benzodiazepine antagonists with antidote properties. While these drugs have relatively low toxicity, dose-related oversedation remains a risk in susceptible patients, especially when combined with other sedatives.


Delayed hypersensitivity skin tests in prognosis of human immunodeficiency virus infection.

Delayed hypersensitivity skin tests (DHST) with recall antigens were investigated as prognostic markers in five different approaches. In the first study, 42 acquired immunodeficiency syndrome (AIDS) patients (IVb, IVcl, IVd, and IVe; MMWR 35:334-339, 1986) 26 AIDS-related complex (ARC) patients (IVa and IVc2), and 98 asymptomatic patients (II and III) were evaluated with candidin, tricophytin, PPD and streptokinase-streptodornase. In the second study, 10 patients (II and III) were evaluated sequentially with the same antigens. In the third, 45 patients with at least two positive skin tests ("reactors") were followed for one year and evaluated every 6 months with the same antigens. In the fourth, 16 "reactors" were followed and evaluated every 3 months with the same antigens. We measured the interval from the time at which patients first presented with only one or no positive DHST until the development of ARC or AIDS. In the last study, the correlation between absolute number of CD4+ lymphocytes and the number of DHST was studied in 151 patients. We found that the decrease in reactiveness to DHST correlated directly with the progression to AIDS, demonstrating the usefulness of this simple procedure as a valid prognostic marker.


Evaluation of ion-exchange microspheres as carriers for the anticancer drug doxorubicin: in-vitro studies.

A comparison study of doxorubicin loading, release characteristics and stability within sodium and hydrogen forms of ion-exchange resin microspheres has been performed. It was demonstrated that resins in the Na+ form, although having lower drug loading capacity, showed similar release profiles to resins in the H+ form but still maintain all the drug activity. Resins in the H+ form, despite having high drug loading capacity, caused drug degradation within microspheres due to their strong acidic nature. Therefore, in comparison with the H+ form, resins in the Na+ form can be considered as better carriers for doxorubicin in terms of sustaining the release of drug and maintaining drug activity. Other factors such as the degree of resin cross-linkage and drug/resin mixing time have also been examined in relation to drug loading and release characteristics. Overall, this study demonstrated the significance of the characteristics of matrix materials and their influence on the drug activity and microsphere performance in-vitro.


Effects of proteolytic enzyme inhibitors as absorption enhancers on the transdermal iontophoretic delivery of calcitonin in rats.

The effects of proteolytic enzyme inhibitors, aprotinin, soybean trypsin inhibitor and camostat mesilate as absorption enhancers on the transdermal iontophoretic delivery of salmon calcitonin (SCT) have been examined in rats. The dermal absorption of SCT was evaluated with hypocalcaemic effect. Application of SCT (12.5 int. units/rat) onto abdominal skin did not produce any hypocalcaemic effect. This produced a small hypocalcaemic effect with cationic iontophoresis (drug phase, anode; reference phase, cathode; high frequency pulses of 1 V at 10 kHz, 2h). Furthermore, camostat mesilate (1 mM) and aprotinin (10(6) int. units mL-1) enhanced the hypocalcaemic effects on the application of SCT with iontophoresis. These hypocalcaemic effects were highest with the pH 4.0 preparation compared with those of the pH 5.5, pH 7.0 and pH 8.0 preparations. However, soybean trypsin inhibitor did not change the hypocalcaemic effects. This was because the soybean trypsin inhibitor is a relatively high molecular weight peptide (mol. wt 8000) and an anion at used pH, and therefore was not absorbed through rat skins with cation iontophoresis.


Effect of a perfluorochemical emulsion on the rat hepatic mixed function oxidase system.

The perfluorochemical components of synthetic oxygen transporting emulsions may persist in hepatic tissue. After a single 30% blood exchange with the perfluorochemical emulsion, Fluosol-DA 20%, the effects on the microsomal metabolism of 7-methoxycoumarin and 7-ethoxycoumarin were studied over a 9-week period. Fluosol-DA treated animals were compared with controls (sham) and hetastarch-treated controls. Changes in dealkylase activities were compared with induction by phenobarbitone and 3-methylcholanthrene. The liver to body weight ratio increased by 49% in Fluosol-DA-treated rats over the controls at 1 week and the microsomal protein was increased in the Fluosol-DA-treated rats after 4 and 9 weeks. Fluosol-DA treatment induced 7-methoxycoumarin demethylase with peak differences occurring at 1 week and a Vmax 75% greater than controls. Fluosol-DA was a more potent inducer of demethylase than phenobarbitone. In addition, 7-ethoxycoumarin de-ethylase was induced by Fluosol-DA with a peak induction at 4 weeks. The Vmax at 4 weeks in Fluosol-DA-treated rats was 122% greater than control. In this case, Fluosol-DA produced less induction in de-ethylase than 3-methylcholanthrene. These studies show that Fluosol-DA induces more than one form of cytochrome P450 and the effects resemble those of phenobarbitone more than those of 3-methylcholanthrene. Hetastarch, a plasma expander, did not affect liver weights, microsomal protein content, or the cytochrome P450 system.


Time course of PAF formation by gastrointestinal tissue in rats after castor oil challenge.

When castor oil was administered by gavage to rats, the duodenum and jejunum, but not the stomach, produced large amounts of platelet activating factor 3-7 h after oil challenge with a peak at 3 h. Intraluminal release of acid phosphatase was also markedly increased in the duodenum and jejunum of castor oil-treated rats, especially 3-5 h after oil challenge. No increase was observed in the stomach. There was a correlation between elevated release of acid phosphatase and intestinal hyperaemia.


Effect of calcium channel blockers on postprandial gastrointestinal motility in the dog.

We have compared the ability of nifedipine and lacidipine, a new 1,4-dihydropyridine, to interfere with postprandial gastrointestinal motility. Five conscious dogs, fitted with 8 bipolar electrodes along the gastrointestinal tract, were studied. Gastrointestinal spike activity was evaluated by means of a computer system. Lacidipine (8 micrograms kg-1) was administered as an i.v. bolus immediately followed by a 10 micrograms kg-1 h-1 i.v. infusion for 3 h, starting 30 min before a standard meal. This dose of lacidipine decreased systolic blood pressure by approximately 20%. Nifedipine was used at equihypotensive doses (30 micrograms kg-1 i.v. bolus followed by 300 micrograms kg-1 h-1 i.v. infusion). Lacidipine had no effect on either gastric or intestinal postprandial spike activity. Nifedipine significantly delayed the appearance of the fed pattern and reduced the number of spikes in the small bowel, while it had no effect on gastric spike activity. We conclude that equihypotensive doses of lacidipine and nifedipine differ in their effects on the gastrointestinal tract, lacidipine having a better cardiovascular selectivity profile than nifedipine, and that the sensitivity to nifedipine varies in different parts of the gut.


Different sites of action for alpha 2-adrenoceptor antagonists in the modulation of noradrenaline release and contraction response in the vas deferens of the rat.

Rat vas deferens was prepared, loaded with [3H]noradrenaline, and superfused to measure the release of tritium in resting conditions and in response to electrical field stimulation. The alpha 2-adrenoceptor antagonists yohimbine, CH-38083 (7,8-(methylenedioxi)-14 alpha-hydroxyalloberbane HCl), and idazoxan increased the electrically induced release of tritium in a concentration-dependent manner, whereas noradrenaline and the alpha 2-adrenoceptor agonist xylazine exerted opposite effects. The inhibitory effect of noradrenaline on electrically induced tritium release was antagonized by yohimbine, CH-38083, and idazoxan. Of the alpha 2-adrenoceptor antagonists tested, yohimbine and CH-38083 reversed the xylazine-induced inhibition of tritium release, and idazoxan was found to be completely ineffective against xylazine. Idazoxan, yohimbine and CH-38083 antagonized the inhibitory effect of xylazine on electrical stimulation-induced contractions of the vas deferens, as was evidenced by the apparent pA2 values. We conclude from the present experiments that noradrenaline and xylazine inhibit noradrenaline release by acting on distinct prejunctional alpha 2-adrenoceptors and that the receptor subtype that responds to xylazine is insensitive to idazoxan. In addition, inhibition by xylazine of contractility but not of noradrenaline release was antagonized by idazoxan, suggesting that besides noradrenergic neurotransmission, other motor transmitter systems (purinergic) may also be involved in the inhibition by alpha 2-adrenoceptor antagonists of mechanical responses in the rat vas deferens.


Effect of cocaine on tritium overflow evoked from vasa deferentia previously loaded with [3H]noradrenaline by stimulation using different types of electrode.

In mouse and guinea-pig vasa deferentia previously incubated with [3H]noradrenaline, electrical stimulation applied through parallel electrodes (transmurally) increased overflow of tritium 2- to 5-fold above the resting value. Electrical stimulation applied using methods involving more substantial conduction of nerve impulses in neuronal elements in the tissues evoked a tritium overflow which was smaller (70%) than that evoked by transmural stimulation. Cinchocaine (25 microM), tetrodotoxin (0.5 microM) or the absence of calcium effectively abolished evoked overflow in both tissues whichever method of stimulation was used. In mouse vas deferens, cocaine (10 microM) did not alter overflow evoked by either transmural or axonal stimulation while 100 microM produced a reduction. In guinea-pig vas deferens, cocaine (10 microM) produced a statistically significant increase in evoked overflow of about 50% or more with both transmural and axonal stimulation. As in mouse vas deferens, 100 microM cocaine produced a reduction. It is concluded that the action of cocaine is independent of these methods of stimulation and that some difference in the arrangement of the noradrenergic nerves in the two species may account for the differential effect of cocaine observed.


Effects of (+/-)-, (+)- and (-)-celiprolol on the rat left atria and portal vein.

Differing effects of (+/-)-celiprolol at beta-adrenoceptors have been reported. The effects of (+/-)-, (+)- and (-)-celiprolol on the contractile responses of the rat left atria and portal vein have therefore been studied. (+/-)-Celiprolol did not augment the responses of the atria to electrical stimulation and is therefore not an agonist at beta 1-adrenoceptors. Prolonged treatment with (+/-)-celiprolol attenuated the contractile activity of the vein and this effect was blocked by ICI 118,551 and is therefore mediated by agonism at beta 2-adrenoceptors. The pA2 values for (+/-)-, (+)- and (-)-celiprolol at the beta 1-adrenoceptors of the atria were 8.0, 6.0 and 8.6, respectively. On the protal vein, (+/-)- and (-)-celiprolol produced non-parallel rightward shifts of log isoprenaline attenuation curves with a reduction in isoprenaline maximum responses. This effect of (+/-)-celiprolol on the vein was not due to slowly reversible antagonism, as the inhibitory effect of (+/-)-celiprolol was readily reversible.


A possible mechanism of endothelium-dependent relaxation induced by pirarubicin and carbachol in rat isolated aorta.

The mechanism of endothelium-dependent relaxation induced by pirarubicin, (2''R)-4'-O-tetrahydropyranyladriamycin, THP, or carbachol was investigated in the rat isolated aorta. The relaxant effect of THP (1.5 x 10(-6)-4.5 x 10(-5) M) or carbachol (10(-8)-10(-4) M) on the aorta with endothelium was decreased by lowering Ca2+ in the medium. The relaxation induced by THP was not inhibited by pretreatment with verapamil (10(-6)-10(-5) M), and that induced by carbachol was only partially inhibited. However, on replacement of all but 20 mM Na+ with either Li+ or choline, the THP- or carbachol-induced relaxation was inhibited. Furthermore, the relaxing effect of THP or carbachol was inhibited by pretreatment with amiloride (10(-4)-3 x 10(-4) M), with ouabain (10(-4)-10(-3) M), or with K(+)-depletion. These results suggest that the THP- or carbachol-induced relaxation depending on endothelium was affected by modifying the calcium ion concentration, and that a Na(+)-Ca2+ exchange process is involved.


Anorectic activity of fluoxetine and norfluoxetine in rats: relationship between brain concentrations and in-vitro potencies on monoaminergic mechanisms.

The present study was aimed at establishing the importance of brain monoamine uptake and release mechanisms in the anorectic activity of fluoxetine, relating them to the actual brain concentrations of the parent drug and its metabolite norfluoxetine after anorectic doses in rats. Both compounds showed anorectic activity when administered intraperitoneally, norfluoxetine being slightly more active (ED50 = 22.9 mumol kg-1) than fluoxetine (ED50 = 35.0 mumol kg-1) despite the fact that the metabolite is about ten times less potent than the parent drug in inhibiting 5-hydroxytryptamine (5-HT) uptake. Comparing the brain concentrations of norfluoxetine, in terms of maximum concentrations (Cmax) and area under the curve (AUC), after the ED50 of fluoxetine or synthetic norfluoxetine, it also appeared that the metabolite plays a major role in the anorectic effect of the parent drug in rats. Brain Cmax of fluoxetine (48.7 microM) and norfluoxetine (21.7 and 27.3 microM after metabolite and drug, respectively) were several times those blocking 5-HT uptake in-vitro (0.5 microM), making it unlikely that fluoxetine (directly or through its metabolite) reduces food intake by specifically blocking 5-HT neuronal uptake. Brain Cmax of fluoxetine but particularly norfluoxetine were more compatible with those capable in-vitro of affecting catecholaminergic mechanisms, such as inhibition of dopamine and noradrenaline uptake and enhancement of dopamine release. These results together with recent in-vitro findings that the parent compound and its active metabolite induce tritium release from hippocampal synaptosomes previously loaded with [3H]5-HT suggest that mechanisms other than inhibition of 5-HT uptake are involved in the anorectic action of these compounds in rats.


Renal clearance-lipophilicity relationships of some organic acids in rabbits, rats and mice.

The effect of lipophilicity on the renal clearance for a group of weak organic acids (benzoic, phenylacetic and hippuric acid derivatives) was studied in rabbits, rats and mice. These compounds are eliminated in the kidney by glomerular filtration and undergo both tubular secretion and tubular reabsorption. For quantification of the effect of lipophilicity, an equation [formula: see text] was employed, where CLR represents renal clearance of the parent drug, ERPF is effective renal plasma flow, D is the partition coefficient of the acids between octanol and water, and a and b are constants. In interspecies comparison, the values of parameters a and b are similar indicating no significant interspecies differences in this route of elimination.


Physiochemical interactions of praziquantel, oxamniquine and tablet excipients.

A differential scanning calorimetry and infrared spectrophotometry study of potential interactions between oxamniquine and praziquantel, two synergistic anti-parasitic drugs, indicated that they did not interact on fusion. Mixtures of the two drugs with each of nine common pharmaceutical excipients (Ac-Di-Sol, Avicel, Crospovidone, calcium phosphate, magnesium stearate, starch, lactose, PEG 6000, stearic acid) appeared to interact only with stearic acid. These results suggested that a combination solid dosage form was feasible.


Physicochemical properties of oxamniquine indicate a new polymorphic form.

New dissolution rate, chemical stability and thermal analysis data are reported for an anti-schistosomal drug, oxamniquine. A slow dissolution rate (40-70% in 1 h) was found, which may contribute to its erratic clinical response. The drug was found to be chemically stable in water for at least 21 days at 37 degrees C. Dissolution rate and thermal analysis evidence is presented for a previously unreported polymorph (Form III). This form appears to be intermediate in physical stability between the known Forms I and II.


Investigation of the 5-HT receptor mediating relaxation in guinea-pig proximal colon.

5-Hydroxytryptamine (5-HT) induced concentration-related relaxations (EC50 = 9.1 microM) of guinea-pig proximal colon pretreated with ketanserin (1 microM) and ondansetron (10 microM). This 5-HT-induced effect was neuronally mediated since it was blocked by tetrodotoxin (0.3 microM). 5-Carboxamidotryptamine was a full agonist and ten times more potent than 5-HT. alpha-Methyl-5-HT was a partial agonist. 2-Methyl-5-HT was without effect. Methysergide and metergoline were antagonists of 5-HT producing parallel shifts at 0.1 microM but unsurmountable antagonism at higher concentrations. pKB values of 8.0 and 7.3 were calculated for methysergide and metergoline, respectively. This study has identified a 5-HT-induced relaxation of guinea-pig proximal colon which is mediated via a neuronal 5-HT1-like receptor. However, the subtype has yet to be established.


Haemolytic action of N-alkylpolymethylenediamines.

The haemolytic action of various N-alkyl derivatives (lauryl; C12H25-, myristyl; C14H29-, palmityl; C16H33-) of 1,3-diaminopropane, 1,4-diaminobutane, 1,5-diaminopentate, 1,6-diaminohexane, 1,7-diaminoheptane, 1,8-diaminooctane was examined using rabbit red blood cells. The activities of the various derivatives were compared with those of antiplaque agents commonly used as mouthwashes; cetylpyridinium chloride (CP) and chlorhexidine acetate (CH). The haemolytic activities of these agents were dependent on the length of the N-alkyl chain, whereas the number of methylene groups between the nitrogen atoms had little effect. The order of potency was CP, N-palmityl derivatives, N-myristyl derivatives greater than N-lauryl derivatives greater than CH which was similar to the order of the antiplaque effect evaluated in-vitro.


The use of albumin microspheres in the treatment of carrageenan-induced inflammation in the rat.

Free hydrocortisone, hydrocortisone incorporated into microspheres and empty microspheres have been administered orally to rats with carrageenan-induced hindpaw inflammation. Hydrocortisone administered in particles was effective at a lower dose than free steroid in reducing inflammation. Inflammatory exudates were able to release steroid from the microspheres by proteolytic degradation.


Effects of morphine: an electrophysiological study on guinea-pig papillary muscle.

Intracellular microelectrodes were used to study the electrophysiological effects of morphine on guinea-pig papillary muscle. Morphine (5 x 10(-4) M) caused a significant decrease in the maximum rate of depolarization. At high concentrations (5 x 10(-3) M) morphine induced a decrease in the action potential amplitude and a prolongation of the action potential duration. The administration of naloxone (10(-7) M) partially antagonized the cardiac electrophysiological effects of morphine. These results suggest that the electrophysiological effects of morphine may be due to an interaction with opioid receptors.


Inhibition by diltiazem of left ventricle collagen proliferation during renovascular hypertension development in rats.

Diltiazem administered in drinking water (0.7 mg mL-1) to Goldblatt two kidney-one clip rats over 16 weeks did not prevent the development of hypertension and left ventricular hypertrophy (LVH). When the collagen content of the left ventricles was assayed (as hydroxyproline), it was found that the fibrosis, characteristic of LVH, was inhibited by diltiazem treatment, despite the fact that hypertension and LVH had developed. This study provides some indirect evidence for the notion that the collagen and myocyte compartments of the myocardium are under separate influences during LVH development in renovascular hypertension.


Peptidergic regulation of the Limulus midgut.

1. The morphology and innervation of the midgut (intestine) in the horseshoe crab, Limulus polyphemus was investigated. The organization of this tissue was examined with routine histology. Radioimmunoassay, immunohistochemistry and high performance liquid chromatography were employed to detect, localize and identify peptidergic innervation of the midgut. The actions of synthetic and native proctolin-like and FMRFamide-like peptides were compared on the isolated midgut preparation. 2. Levels of proctolin and FMRFamide were determined in extracts of Limulus midgut tissue using radioimmunoassay. High levels of proctolin-like immunoreactivity (69.5 +/- 11.3 ng/g) were detected, while levels of FMRFamide-like immunoreactivity (0.8 +/- 0.2 ng/g) were less. Proctolin levels were equally distributed, while the levels of FMRFamide-like immunoreactivity exhibited an anterior bias. 3. Proctolin- and FMRFamide-like immunoreactivities in the Limulus midgut were localized with immunohistochemistry. Proctolin- and FMRFamide-immunoreactive elements were detected in intestinal nerve branches and individual fibers running along the surface of the midgut in whole-mount preparations. In sectioned tissue, staining for these peptides was observed throughout the midgut, typically associated with muscle bands and fibers. Only a few immunoreactive cell bodies were observed. 4. Proctolin, and several FMRFamide-like peptides produced distinct and opposing actions on the isolated Limulus midgut preparation. Proctolin elicited contracture and rhythmic contractions of this tissue, while FMRFamide and N-terminally extended analogs of FLRFamide relaxed gut tension. FMRFamide-like peptides partially reversed the excitatory actions of proctolin. 5. Proctolin- and FMRFamide-like peptides in Limulus midgut extracts were partially characterized with high performance liquid chromatography. One peak of proctolin-like activity was detected on a linear gradient of 18 to 31.5% acetonitrile. The native proctolin-like peptide produced excitatory actions on the isolated midgut preparation which were indistinguishable from those produced by synthetic proctolin. Several peaks of FMRFamide-like bioactivity (Busycon radula protractor muscle assay) were detected with a linear gradient of 5 to 30% acetonitrile. Fractions from two distinct peaks produced FMRFamide-like inhibitory effects on the isolated Limulus midgut preparation. These findings suggest a role for proctolin-like and FMRFamide-like peptides as regulators of intestinal motility in Limulus.


Labelling of rat brain beta-adrenoceptors: (3H)CGP-12177 or (125I)iodocyanopindolol?

Binding of (125I)iodocyanopindolol (ICYP) and (3H)CGP-12177 to rat brain homogenates was characterized and compared. ICYP was shown to bind to both beta-adrenergic and serotonin1B (5HT1B) receptors whereas (3H)CGP-12177 only labelled the first ones. The addition of 10 microM serotonin (5HT) prevented ICYP binding to 5HT receptors and under these experimental conditions both ligands labelled a similar total number of beta-adrenoceptors in the different rat brain regions. ICYP displayed a higher affinity for cerebellar (mainly beta 2-subtype) than for cerebral cortex beta-adrenoceptors (mainly beta 1-subtype) suggesting a subtype selectivity. A multiple displacement binding approach using CGP-20712A, a beta 1-subtype ligand, as competitor revealed a 2.6 fold selectivity of ICYP for the beta 2-adrenoceptor subtype. On the other hand, (3H)CGP-12177 binds only to beta-adrenoceptors and is not subtype selective in the rat brain homogenate. Considering both its high specificity and its lack of subtype selectivity (3H)CGP-12177 seems to be a more suitable ligand than ICYP to non-selectively label beta-adrenoceptors in rat brain.


Prenatal care and delivery in an agoraphobic woman. A case report.

Agoraphobia is a psychiatric illness that can cause patients to become functionally homebound. Although the obstetric care of women with agoraphobia has not been described previously, it is likely that the problem is more prevalent than recognized. A pregnant woman initially believed she would be unable to travel to the hospital for delivery. She was given prenatal care and psychotherapy at home; a safe hospital delivery was achieved. The management plan included the in-depth involvement of two physicians, physician preparation for a potential home delivery, patient education about obstetric risks and the harmlessness of agoraphobic panic attacks, and psychologic and marital therapy. Strategies for minimizing medicolegal risks were employed.


N-(5,5-diacetoxypent-1-yl)doxorubicin: a new intensely potent doxorubicin analogue.

N-(5,5-Diacetoxypent-1-yl)doxorubicin (DAPDOX) (3), a new, water-soluble analogue of doxorubicin, has been synthesized by coupling doxorubicin with 5-oxopentane-1,1-diacetate in the presence of NaBH3CN. This analogue was designed to be converted to the corresponding aldehyde, N-(5-oxopent-1-yl)doxorubicin, in the presence of carboxylate hydrolases, enzymes that are ubiquitous in tissue. DAPDOX had a half-life of several days in 0.05 M phosphate or 0.05 M acetate buffer solution at pH 4.0. However, in 0.05 M phosphate buffer at pH 7.4 in the presence of 20 unit equiv of porcine liver carboxylate esterase, the half-life of DAPDOX was less than 1 min. N-(5-acetoxypent-1-yl)doxorubicin (4), which should give rise to N-(5-hydroxypent-1-yl)doxorubicin on esterase-mediated hydrolysis, and N-(pent-1-yl)doxorubicin (5), were also prepared for comparative biological studies. DAPDOX was 150 times more potent than doxorubicin at inhibiting the growth of Chinese hamster ovary (CHO) cells in culture. The compound retained the same degree of potency against a CHO subline 100-fold resistant to doxorubicin (CHO/DOX) that expressed elevated levels of P-glycoprotein. Compounds 4 and 5, on the other hand, were no more effective than doxorubicin at inhibiting the growth of CHO cells and were 4-7-fold less potent against the CHO/DOX subline. DAPDOX is representative of a new structural class of doxorubicin analogues with unique chemical and biological properties.


Novel thiosemicarbazones derived from formyl- and acyldiazines: synthesis, effects on cell proliferation, and synergism with antiviral agents.

The synthesis of a series of novel thiosemicarbazones (TSC's) derived from various alkyl diazinyl (3-pyridazinyl, 4-pyrimidinyl, 2-pyrazinyl) ketones and 3-pyridazinecarbaldehyde and their evaluation against herpes simplex virus (HSV) and human immunodeficiency virus (HIV) as well as the determination of their cytotoxicity are described. In addition, the effects of combination of such TSC's with the well-known antiviral drugs acyclovir (ACV) and 3'-azido-3'-deoxythymidine (AZT) were studied. Under our experimental conditions, i.e. determination of virus-induced cytopathic effect upon infection of HUT78 cells with HSV-1 and upon infection of MT4 cells with HIV-1, no antiviral activity could be detected with any of the TSC's. However, pronounced effects on proliferation of these rapidly growing T4 lymphocyte cell lines were observed. Clear structure-activity relationships with regard to these cytotoxic effects could be established: compared to pyridine, pyrazine, or pyrimidine-derived TSC's most of the 3-pyridazinyl congeners investigated are less cytotoxic; introduction of a methyl group into C-6 of the pyridazine system or prolongation of the acyl moiety in these compounds has essentially no influence; all compounds bearing an N,N-dimethylamino or a cycloamino substituent are much more toxic than those with an NH2 or NHR substituent; the nature of R in the latter type of compounds has only moderate influence. It has been reported that combination of TSC's with the antiviral agent acyclovir (ACV) results in potentiation of this well-known drug. We evaluated the potential of our series of novel TSC's in combination with ACV for inhibition of HSV-1-induced cytopathic effect in HUT78 cells and in combination with 3'-azido-3'-deoxythymidine (AZT) for inhibition of HIV-1-induced cytopathic effect in MT4 cells. Only four compounds out of this series, all characterized by an unsubstituted NH2 group, exhibited moderate synergism with the above mentioned antiviral drugs. Our results do not support the previously expressed opinion that TSC's are selective antiviral agents. In our test systems no evidence for inhibition of virus-induced cytopathic effect was obtained. The TSC derivatives exhibited a broad range of cytotoxic effects, some at concentrations considerably below those reported to have antiviral efficacy. Several of our novel diazine-derived compounds proved advantageous over the previously described pyridine analogues with regard to cytotoxicity. Moderate synergism could be detected for relatively noncytotoxic TSC's with the antiviral drugs ACV (antiherpes) and AZT (anti-HIV).


Development of renal toxicity in F344 rats gavaged with mercuric chloride for 2 weeks, or 2, 4, 6, 15, and 24 months.

Both sexes of F344 rats were gavaged with maximal tolerated doses of mercuric chloride for periods from 2 wk to up to 2 yr to investigate chronic nephrotoxicity and potential carcinogenicity. The toxicity of mercuric chloride was excessive after 2 wk of exposure to doses ranging from 1.25 to 20 mg/kg, compromising renal function by selectively destroying cells of the proximal tubules, and eliciting marked elevations in urinary biomarker enzymes diagnostic for acute renal tubule necrosis. In the 2-wk studies, urinary alkaline phosphatase and aspartate amino-transferase were most sensitive to renal mercury toxicity among a panel of six enzymes, exhibiting twofold increases above controls at the 5.0 mg/kg dose, before changes in the other enzymes occurred. Urinary lactate dehydrogenase was the most responsive enzyme, with up to 11-fold increases in activity above controls. In response to mercuric chloride exposure of 5.0 mg/kg for 2-6 mo, the greatest and most persistent increases in elevation of urinary enzyme activities were exhibited by alkaline phosphatase and gamma-glutamyl transferase, which increased two-to threefold above controls. At this interval, the maximal severity of the renal lesions in both sexes of rats was graded as minimal to mild. Beyond 6 mo none of the urinary enzymes measured in this study was adequate as biomarkers of nephrotoxicity, although the severity of the renal lesions had progressed. Mercury accumulated in a dose-related fashion primarily in the kidney, and to a lesser extent in the liver. The severity of the renal lesions was increased by continued exposure to mercuric chloride, as tissue concentrations of mercury rose in proportion to dose. Mercuric chloride treatment for 2 yr clearly exacerbated the severity of the spontaneous nephrotoxicity prevalent in aging F344 rats. The excessive mortality that occurred in the male rats was probably due to a combination of these factors. No renal tumors were detected in rats, possibly because the potential for their development was reduced; however, direct tissue contact with mercury induced squamous-cell papillomas of the forestomach in both sexes.


Does transurethral laser ureterolithotripsy justify its cost?

A prospective consecutive series of 64 patients who underwent transurethral laser ureterolithotripsy using a 7.2F semirigid ureteroscope was compared to the immediately preceding consecutive series of 98 patients who had undergone ultrasound lithotripsy using rigid 9.5F or 12.5F ureteroscopes. The distribution of the calculi by size and composition in both series was similar. There was a higher proportion of upper ureteral calculi in the laser lithotripsy series. The success rate for a first attempt at laser lithotripsy was 92.2% versus 71.4% for the ultrasound series (p less than 0.01). When the stone could be reached ultrasound and laser lithotripsy had a fragmentation rate of 97%. The principal reason for the difference in results was the poorer ability to reach calculi when using the larger rigid ureteroscopes. One patient who had failed ultrasound lithotripsy was successfully treated with laser lithotripsy a year later. The overall morbidity was less for laser lithotripsy. The 3-year cost-benefit analysis revealed a smaller difference in cost than expected and the 5-year analysis was advantageous for laser lithotripsy because of its higher success rate. Savings were also realized in the laser series because of the higher proportion of subjects treated as outpatients, and a lower mean duration of hospitalization and time missed from work. For our center with an annual work load of approximately 100 cases laser lithotripsy achieved a superior cost-benefit ratio.


[Clinical applications of ion-sensor].

The coulometric measurement technique offers a rapid, accurate and precise method for measuring iron in liquid samples. It is well-known that analytical interferences common to colorimetric methods essentially do not exist in this procedure. However, this study shows that various drugs, such as chondroitin iron sulfate, deferoxamine mesylate, and salazosulfapyridine have strongly positive effects on the most contemporary determination of serum iron. The study of the analytical interference of drugs is important for precise and accurate measurement of desirable or undesirable pharmacological effects. New drugs and new methods should be systematically studied in this manner.


L-365,260, a potent CCK-B/gastrin receptor antagonist, suppresses gastric acid secretion induced by histamine and bethanechol as well as pentagastrin in rats.

We evaluated the effects of a potent cholecystokinin (CCK)-B/gastrin receptor antagonist, L-365,260 (3R(+)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin - 3-yl)-N'-( 3-methylphenyl) urea); a selective CCK-A receptor antagonist, devazepide (L-364,718); and cimetidine on gastric acid secretion induced by pentagastrin, histamine and bethanechol in anesthetized rats. We also evaluated the effects of L-365,260 and cimetidine on acid secretion in pylorus-ligated rats. Intravenous administration of L-365,260, L-364,718 and cimetidine dose-dependently reduced acid secretion induced by pentagastrin (20 nmol/kg/hr), with ED50 values of 0.63, 19.1 and 2.5 mumol/kg, respectively. Of interest was the finding that L-365,260, like cimetidine, dose-dependently inhibited acid secretion induced by histamine (100 mumol/kg/hr) and bethanechol (5 mumol/kg/hr) with ED50 values of 5.9 and 4.3 mumol/kg, respectively. L-364,718, even at 30 mumol/kg, i.v., had only a slight effect on histamine- or bethanechol-induced acid secretion. Gastric acid secretion was suppressed by treatment with L-365,260 (3-100 mumol/kg, i.v.) and cimetidine (11.9-396.4 mumol/kg, i.v.) in pylorus-ligated rats, with ED50 values of 13.3 and 96.9 mumol/kg, respectively. These results indicate that L-365,260 suppresses acid secretion induced by histamine and bethanechol in rats and that the gastrin receptor plays an important role in acid secretion in pylorus-ligated rats.


Selectivity of bevantolol hydrochloride towards alpha- and beta-adrenoceptor subtypes in rat cerebral cortex.

Selectivity of bevantolol hydrochloride (NC-1400) towards alpha- and beta-adrenoceptor subtypes of rat cerebral cortex was examined in binding experiments and compared with propranolol. Bevantolol biphasically displaced the 3H-dihydroalprenolol binding. The affinity of bevantolol to beta 1-adrenoceptor was equal to that of propranolol. Bevantolol displaced 3H-prazosin binding monophasically but not 3H-p-aminoclonidine binding. These results suggest that bevantolol is a beta 1-adrenoceptor antagonist with a relatively high affinity to alpha 1-adrenoceptor subtypes.


Serotonergic mechanisms in anxiolytic effect of tandospirone in the Vogel conflict test.

To clarify which 5-HT1A receptors, autoreceptors located in the raphe nuclei or post-synaptic receptors in the forebrain areas receiving a 5-HT input, mediate the anticonflict action of tandospirone (a 5-HT1A receptor-related anxiolytics), the behavioral effects of tandospirone were studied in 5,7-dihydroxytryptamine (5,7-DHT) treated rats. By measuring both monoamines and their metabolite levels and densities of [3H]8-OH-DPAT binding in 5,7-DHT-treated rat brain, we confirmed that pretreatment with 5,7-DHT destroyed 5-HT neurons selectively without affecting postsynaptic 5-HT1A receptors located on the postsynaptic neurons. This selective destruction produced no significant changes in the drinking behavior of rats in either punished or unpunished sessions of the Vogel conflict test. Furthermore, this destruction altered neither the effect of tandospirone on punished responding in this procedure nor the potency of tandospirone to induce a flat body posture in rats, which is known as the "serotonin behavioral syndrome". These results suggested that the anticonflict action of tandospirone may be produced, at least in part, by binding to postsynaptic 5-HT1A receptors and activating them as agonists, and not to 5-HT1A autoreceptors located on the cell bodies of 5-HT neurons.


Sympathetic nerve stimulation activates both beta 1- and beta 2-adrenoceptors of SA and AV nodes in anesthetized dog hearts.

We investigated blocking effects of the selective beta 1-adrenoceptor blocker atenolol (0.1-100 micrograms/kg, i.v.), the selective beta 2-adrenoceptor blocker ICI 118,551 (1-1000 micrograms/kg, i.v.) and the combination of the two drugs on positive chronotropic and dromotropic responses to norepinephrine (NE) released by stimulation of the sympathetic nerves in anesthetized, neurally decentralized, open-chest dogs after atropine was given. Stimulation of the intracardiac sympathetic nerves to the SA nodal region or to the AV nodal region selectively increased heart rate or decreased AV conduction time, respectively. ICI 118,551 inhibited the chronotropic or dromotropic response to each stimulation in a dose-dependent manner, but its inhibition of the dromotropic response was less than that of the chronotropic response. Atenolol similarly inhibited either the positive chronotropic or dromotropic response to each stimulation in a dose-related manner. The combination of atenolol and ICI 118,551 attenuated the responses to each stimulation more than atenolol alone. These data indicate that sympathetic nerve stimulation activates both beta 1- and beta 2-adrenoceptors of the SA and AV nodes and that the proportion of beta 2-adrenoceptor-mediated effects on the AV node is less than that on the SA node. These results suggest that neurally released NE in part controls physiological functional cardiac responses mediated through beta 2-adrenoceptors, in addition to the responses predominantly mediated through beta 1-adrenoceptors.


[Differentiation and maturation of excurrent duct system of rat testes].

The present study was designed to investigate relations of gamma-Glutamyl transpeptidase (gamma-GTP) activities to morphological differentiation and maturation of rat testicular excurrent duct system including rete testis, efferent ductuli and epididymal ducts. Animals used were Wistar rats aged from 3 days to 12 weeks. Histochemical demonstration of gamma-GTP activities was carried out by the method of Rutenburg et al. (1969). Morphological differentiation of the epithelium and structure of the rete testis occurred between 2 and 3 weeks and completed by the age of 6 weeks, which was accompanied by induction and deletion of gamma-GTP activity. Morphological differentiation and maturation of the epithelium of the efferent ductuli and the epididymal ducts became evident by the age of 4 weeks and gamma-GTP was active in both epithelial cells throughout the period examined. Since the maturation of rete testis epithelium was found to proceed as observed as Sertoli cells, the origin and functions of the rete testis epithelium and Sertoli cells are considered to be identical. The simultaneous development of the efferent ductuli and the epididymal ducts suggests that both tissues originate from mesonephric ducts and have similar functions, although mature epithelial cells of both tissues are completely different.


[Pulmonary edema in patients with hemorrhagic fever with renal syndrome].

Eight patients suffering from hemorrhagic fever with renal syndrome (HFRS) running a severe course complicated by pulmonary edema developed absolute hyperhistaminemia and hyperserotoninemia, histamine and serotonin accumulation in tissues. These amines inactivation in blood and lungs and excretion of catecholamines with urine got disturbed. High blood and lung tissue levels of biologically active substances resultant in enhanced permeability of the vascular wall and alveolar epithelium, hemodynamic disturbances due to hypoexcretory hypercatecholaminemia are thought to underlie the occurrence of this grave HFRS complication.


Measurement of free cytosolic calcium in single cells: method and application.

Intracellular calcium [Ca2+]i acts as an important intracellular messenger system for secretion and synthesis, cell growth and differentiation. In order to demonstrate definitively that a change in [Ca2+]i is responsible for a physiological event, one has to measure [Ca2+]i directly within intact cells and correlate the time course of any [Ca2+]i changes with the biological response. Measurement of [Ca2+]i was done in a single cell preloaded with fluorescent Ca indicator fura2 using a fluorescent unit (lonoquant) consisting of an inverted microscope (Zeiss IM 35) equipped with a mercury lamp and a rotating filter wheel containing filters at wavelengths of 340 and 380 nm. Cells were alternately excited and emission signals of fura 2-loaded cells were collected by a photomultiplier and recorded on-line on a computer screen. As a model system, the rat C-cell carcinoma cell line rMTC 6-23 secreting calcitonin was used. An acute elevation of extracellular calcium resulted in an increase in [Ca2+]i within 5 sec and rapid release of preformed calcitonin. This tight linkage between extracellular calcium and [Ca2+]i is mediated via Ca influx through voltage-dependent Ca channels. These channels are modulated by intracellular cAMP, yielding a rhythmic oscillation of [Ca2+]i, as well as by extracellular somatostatin blocking the Ca channel and the increase of [Ca2+]i via a pertussis toxin sensitive Gi protein. The change in [Ca2+]i is associated with changes in calcitonin secretion, confirming the stimulus secretion coupling via voltage-dependent Ca channels in C-cells.


Coeliac disease, epilepsy, and cerebral calcifications. The Italian Working Group on Coeliac Disease and Epilepsy.

There have been anecdotal reports of an association between coeliac disease and epilepsy with cerebral calcifications that resemble those of the Sturge-Weber syndrome. A series of patients who had epilepsy with calcifications, in whom coeliac disease (CD) was incidentally observed, prompted us to study this association. 43 patients (15 male, age range 4.6-30.7 years) were selected from two series. 31 patients with cerebral calcifications of unexplained origin and epilepsy (series A) underwent intestinal biopsy. 12 patients with CD and epilepsy (series B) underwent computed tomography. Antibodies to gluten, folic acid serum concentrations, were measured, and HLA typing was done in most patients. 24 of the series A patients were identified as having CD on the basis of a flat intestinal mucosa (15/22 with a high concentration of serum antigluten), and 5 series B patients showed cerebral calcifications, giving a total of 29 cases with the combination of CD, epilepsy, and cerebral calcifications (CEC). In 27 of these CEC patients, calcifications were located in the parieto-occipital regions. Only 2 of the series A patients had gastrointestinal symptoms at the time of intestinal biopsy; most patients had recurrent diarrhoea, anaemia, and other symptoms suggestive of CD in the first 3 years of life. The epilepsy in CEC patients was poorly responsive to antiepileptic drugs. Gluten-free diet beneficially affected the course of epilepsy only when started soon after epilepsy onset. Cases of "atypical Sturge-Weber syndrome" (characterised by serpiginous cerebral calcifications and epilepsy without facial port-wine naevus) should be reviewed, and CD should be ruled out in all cases of epilepsy and cerebral calcifications of unexplained origin.


Primary pancreatic beta-cell secretory defect caused by mutations in glucokinase gene in kindreds of maturity onset diabetes of the young.

Maturity-onset diabetes of the young (MODY), characterised by non-insulin-dependent diabetes mellitus (NIDDM) with an early age of onset, is a genetically heterogeneous disorder. In most MODY kindreds described in France, chronic hyperglycaemia is caused by mutations in the gene encoding pancreatic beta-cell and liver glucokinase (GCK). We here report the beta-cell secretory profiles of nine patients from four GCK-linked MODY kindreds. First-phase insulin secretion assessed by an intravenous glucose test was comparable in patients and seven controls. However, beta-cell secretory response to continuous glucose stimulus during a hyperglycaemic glucose clamp was significantly reduced: mean plasma insulin values of 12 (SD 7) vs 40 (11) mU/l (p = 0.0001) and mean plasma C-peptide values 1.20 (0.30) vs 2.61 (0.37) (p = 0.0001). This secretory profile is different from those for NIDDM with late age of onset or MODY not linked to GCK. Fasting plasma insulin and C-peptide in patients were inappropriately low in relation to concomitant plasma glucose level. Furthermore, during a hyperinsulinaemic euglycaemic clamp, endogenous insulin secretion at euglycaemia (5 mmol/l) was suppressed in patients but not in controls. These results suggest that mutant GCK may lead to chronic hyperglycaemia by raising the threshold of circulating glucose level which induces insulin secretion. These data provide the first demonstration of a primary pancreatic secretory defect associated with a form of NIDDM.


Immunodeficiency presenting as hypergammaglobulinaemia with IgG2 subclass deficiency.

Recurrent bacterial infections, lymphadenopathy, and failure to thrive are unlikely to be attributed to immune deficiency if they occur in the presence of hypergammaglobulinaemia, and other explanations will usually be sought. We describe eight patients who presented with all these features in infancy or early childhood. Deficiencies of immunoglobulin and antibody production were initially discounted, and the children were referred for investigation of possible lymphoma, autoimmune disease, or chronic viral infection. The patients were later referred to us for more detailed immunological investigation, which revealed low levels of IgG2 and poor specific antibody production to common pathogens. Treatment with intravenous immunoglobulin resulted in resolution of signs and symptoms in all patients. Thus we have shown that hypergammaglobulinaemia does not preclude the presence of immunoglobulin/antibody deficiency. We suggest that investigation of children with high levels of IgG and features of immunodeficiency should include IgG subclass analysis.


Filovirus clearance in non-human primates.

There has been concern in the USA and Europe about filovirus outbreaks in recently imported monkeys, and possible transmission to human beings. Healthy monkeys have been found to have low-titre immunofluorescence antibody (IFA) to Asian filoviruses (Reston and Pennsylvania viruses) as well as to the African filoviruses that caused fulminating human outbreaks in the 1970s (Ebola [Zaire] and Sudan viruses). We have assessed whether such monkeys are a risk to man. We studied 42 non-human primates; 31 were experimentally infected with African and Asian filoviruses, 6 were infected during a documented Reston filovirus outbreak, and 5 had serological evidence suggestive of recent filovirus infection. During the first 15 days after infection, virus could be routinely recovered from serum or biopsy or necropsy tissue, and Asian filovirus RNA could be detected by polymerase chain reaction. 20 to 600 days after challenge, filovirus could no longer be recovered nor viral RNA detected in 141 serum, liver, spleen, or kidney specimens. Animals surviving filovirus infection develop high-titre, cross-reacting filovirus-specific antibody 14 to 21 days after infection, and this coincides with virus clearance. Healthy monkeys with low-titre filovirus antibody may be regarded as uninfected.


Low cerebrospinal-fluid concentrations of soluble amyloid beta-protein precursor in hereditary Alzheimer's disease.

In Alzheimer's disease, deposits of amyloid beta-protein are apparently derived from intracellular processing of a large precursor protein. We have measured concentrations of this precursor in cerebrospinal fluid (CSF) from six members of a family affected by presenile Alzheimer's disease associated with a point mutation of the precursor gene. One gene carrier with clinical signs of the disorder had low CSF concentrations of the precursor, similar to those of three patients with sporadic Alzheimer's disease subsequently confirmed at necropsy. Two symptom-free gene carriers had CSF precursor concentrations similar to those of non-demented controls, though the value was lower in one, who had deficits revealed on neuropsychological testing, than in the other. These findings suggest that low concentrations of soluble amyloid precursor proteins in the CSF reflect the process that results in amyloid plaque formation and vascular deposition in Alzheimer's disease.


Pathogenesis of non-insulin-dependent diabetes mellitus in the black population of southern Africa.

Non-insulin-dependent diabetes mellitus (NIDDM) is an important health problem in the black population of southern Africa. Whether the primary cause of NIDDM is insulin secretory dysfunction or peripheral insulin resistance is unknown. In westernised populations it is believed that insulin resistance and hyperinsulinaemia occur in the early stages of disease, followed later by progressive impairment of insulin secretion. However, we suggest that in the southern African black population a decrease in the mass of functioning beta cells is an important event, making these people vulnerable to the deleterious effects of insulin resistance induced by obesity and other factors. These abnormalities are, in turn, associated with insulin receptor down-regulation. An accelerated decline in beta-cell function then follows in susceptible individuals, ultimately producing striking insulinopenia. Insulinopenic NIDDM in black southern Africans may partly explain why this population has a comparatively low incidence of macrovascular complications and also predicts a short-lived therapeutic response to oral sulphonylureas in most patients.


Impact of user fees on attendance at a referral centre for sexually transmitted diseases in Kenya.

We investigated the impact of a short-lived policy of charging fees to patients attending public-sector outpatient health facilities in Kenya by collecting data on attendance at Nairobi's Special Treatment Clinic for sexually transmitted diseases (STDs) before (23 months), during (9 months), and after (15 months) the user-charge period. During the user-charge period, the seasonally adjusted total mean monthly attendance of men decreased significantly to 40% (95% CI 36-45) of that before fees were levied. Attendance rose in the post-user-charge period, but reached only 64% (59-68) of the pre-user-charge level. For women, the adjusted total mean monthly attendance during the user-charge period was reduced significantly to 65% (55-77) of the pre-user-charge level. Mean monthly attendance by women rose in the post-user-charge period to 22% (9-37) above the pre-user-charge level. There was no evidence of an increase in attendance over the course of the user-charge period among either men or women. The introduction of user fees probably increased the number of untreated STDs in the population, with potentially serious long-term health implications. The user-fee experience in Kenya should be carefully evaluated before similar measures are introduced elsewhere.


The role of amyloid beta-protein in Alzheimer's disease.

Deposition of amyloid beta-protein in the brain has been regarded as the central event in Alzheimer's disease; however, amyloid beta-protein precursor is an endogenous protein, probably with neurotrophic functions. An alternative hypothesis is that amyloid beta-protein is involved in the disease process secondarily, as a protective reactant when brain cells are injured. Insufficiency of amyloid beta-protein precursor, whether because of a genetic defect or in relation to the action of environmental factors, then allows development of Alzheimer changes.


The POT1 gene for yeast peroxisomal thiolase is subject to three different mechanisms of regulation.

The Saccharomyces cerevisiae POT1 gene is, as are other yeast peroxisomal protein genes, inducible by fatty acids and repressible by glucose. We have now found that it is also induced during the stationary phase of the culture. To investigate these three regulatory circuits, we have studied the mRNA levels of regulatory mutants as well as the changes in chromatin structure upon gene activation. We conclude that the regulation of transcriptional activity in glucose repression, oleate induction, and stationary phase induction follow different molecular mechanisms. We suggest that this multiplicity of regulatory mechanisms may represent a general rule for the yeast peroxisomal protein genes.


PulO, a component of the pullulanase secretion pathway of Klebsiella oxytoca, correctly and efficiently processes gonococcal type IV prepilin in Escherichia coli.

The PulO protein required for extracellular secretion of pullulanase by Klebsiella oxytoca is known to be highly homologous to two type IV prepilin peptidases, namely XcpA(PilD) (Pseudomonas aeruginosa) and TcpJ (Vibrio cholerae). The predicted prepilin peptidase activity of PulO was confirmed by showing that it could correctly process the product of the cloned pilE.1 type IV pilin structural gene from Neisseria gonorrhoeae in Escherichia coli. The P. aeruginosa prepilin peptidase and another putative prepilin peptidase, ComC from Bacillus subtilis, also processed prePilE. Subcellular fractionation showed that the pilE gene product that had been processed by PulO remained associated with the cytoplasmic membrane, as did the unprocessed precursor. PulO was also shown to process three of the four prePilE-PhoA hybrids tested. Southern hybridization experiments suggest that a pulO homologue is present in the N. gonorrhoeae chromosome.


Mycobacteria contain two groEL genes: the second Mycobacterium leprae groEL gene is arranged in an operon with groES.

In contrast to other bacterial species, mycobacteria were thus far considered to contain groEL and groES genes that are present on separate loci on their chromosomes, Here, by screening a Mycobacterium leprae lambda gt11 expression library with serum from an Ethiopian lepromatous leprosy patient, two DNA clones were isolated that contain a groEL gene arranged in an operon with a groES gene. The complete DNA sequence of this groESL operon was determined. The predicted amino acid sequences of the GroES and GroEL proteins encoded by this operon are 85-90% and 59-61% homologous to the sequences from previously characterized mycobacterial GroES and GroEL proteins. Southern blotting analyses with M. leprae groES- and groEL-specific probes demonstrate that similar groESL homologous DNA is present in the genomes of other mycobacteria, including Mycobacterium tuberculosis. This strongly suggests that mycobacteria contain a groESL operon in addition to a separately arranged second groEL gene. Using five T-cell clones from two leprosy patients as probes, expression of the M. leprae GroES protein in Escherichia coli after heat shock was demonstrated. Four of these clones recognized the same M. leprae-specific GroES-derived peptide in a DR2-restricted fashion. No expression of the groEL gene from this operon was detected in E. coli after heat shock, as tested with a panel of T-cell clones and monoclonal antibodies reactive to previously described GroEL proteins of mycobacteria.


Positive and negative cis-regulatory elements in the bithoraxoid region of the Drosophila Ultrabithorax gene.

The Ultrabithorax (Ubx) gene is required during embryogenesis and larval development to specify the third thoracic and first abdominal segments of Drosophila melanogaster. Mutations in the bithoraxoid (bxd) region, a 40 kb DNA stretch upstream of the Ubx promoter, affect cis-regulatory elements responsible for the ectodermal expression of the Ubx gene in the posterior compartment of the third thoracic segment and anterior compartment of the first abdominal segment. Our genetic data and the available molecular information are used to map the adult epidermal cis-regulatory elements within the bxd region. Genetic combinations involving mutations affecting the bxd region show that (1) redundant or cooperatively acting sequences are required for Ubx gene expression in the anterior compartment of the first abdominal segment, and (2) the expression of Ubx in the posterior compartment of the third thoracic segment is modulated by positive and negative cis-regulatory elements.


The acu-1 gene of Coprinus cinereus is a regulatory gene required for induction of acetate utilisation enzymes.

We have isolated a gene from Coprinus cinereus which cross-hybridises to the facA and acu-5 genes of Aspergillus nidulans and Neurospora crassa, respectively. These genes encode acetyl-CoA synthetase, an enzyme which is inducible by acetate and required for growth on acetate as sole carbon source. We have designated the C. cinereus gene acs-1 and have used transformation to demonstrate its functional homology to the ascomycete genes by complementation of an N. crassa acu-5 mutation. The acs-1 gene has never been identified by mutation; mutations leading to loss of acetyl-CoA synthetase function map to another gene, acu-1. Using Northern analyses we have shown that acu-1 has a regulatory function that is required for acetate-induced transcription of acs-1 and of another acetate utilisation gene, acu-7, the isocitrate lyase structural gene.


[Age-related changes in mosquito insecticide resistance and their relation to the mechanisms of detoxication].

The mechanisms of DDT, permethrin and malathion resistance were determined indirectly by means of synergists inhibiting the detoxification enzymes. It was found that metabolic resistance dependent on glutathione-S-transferase, MFO and carboxylesterase is the most liable to age changes. Such changes are less expressive in mosquitos which have supposedly kdr-resistance. In young mosquitos (1-2 days) of all species, the rate of synergism is relatively low and fails to explain high resistance which is usual for juvenile specimen.


Human glucokinase gene: isolation, structural characterization, and identification of a microsatellite repeat polymorphism.

The gene encoding human glucokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1), a major component of glucose sensing in pancreatic islet beta-cells, was isolated and characterized. The gene was shown by Southern blotting to exist as a single copy in the genome which mapped to chromosome 7p. It contained 12 exons including two tissue-specific first exons, one active in islet beta-cells (1B), and the other active in liver (1H), and one optional cassette exon which was expressed as a minor form in the liver. Thus the three previously reported isoforms of glucokinase mRNA were the result of tissue-specific activation of separate liver and islet promoters and subsequent alternative splicing events. Eleven exons, including 1H and the optional cassette exon 2A, were scattered over 16 kilobase (kb) in the genome, while exon 1B was separated from the rest by at least 20 kb. Although the islet promoter was found to lack a TATA box, a major transcript from the islet promoter was mapped 486 nucleotides upstream of the translation initiation site. The presence in the islet glucokinase promoter of the potential control element GCCACCAG, a homology of the regulatory element present in both human insulin (GCCACCGG) and rat insulin (GCCATCTG) genes, implied a possible tissue-specific regulatory role of this element. The liver promoter was found to contain a TATA box-like sequence, and transcription was initiated predominantly at 168 nucleotides upstream of the translation initiation site of the major isoform. A new highly polymorphic microsatellite, composed of a compound imperfect dinucleotide repeat [GT]15[GA]8CA[GA]7CA[GA]3AA[GA]2, was mapped 6 kb upstream of islet exon 1. A polymerase chain reaction-based assay was developed, and seven different sized alleles were identified in American Blacks. The sequence information reported here, along with the new polymorphic marker, will make it possible to clarify the molecular basis of potential glucokinase defects in noninsulin-dependent diabetes mellitus patients and may further elucidate the nature of genetic susceptibility to the development of this common metabolic disease.


[The efficacy of a combined preparation based on Bacillus sphaericus and Bac. thuringiensis H-14 against the larvae of blood-sucking mosquitoes].

The optimal bactoculicide/sphaerolarvicide ratio in combination using against Aedes aegypti larvae was established in laboratory by selection. The field trials of the selected BS/BT sample containing the ingredients in titer: bactoculicide-83, sphaerolarvicide-17 were conducted in the arid area of Central Asia. With 1 g/m2, complete mortality of Culex, Aedes and Anopheles larvae was obtained with the residual larvicidal action of 15 days. It is recommended to carry out more large-scale trials of the preparation against Anopheles larvae in different regions.


Heterologous desensitization of the human dopamine D1 receptor in Y1 adrenal cells and in a desensitization-resistant Y1 mutant.

In previous studies, mutant clones (designated Y1DR) were isolated that resisted ACTH-induced homologous desensitization of adenylyl cyclase. The Y1DR mutation also conferred resistance to the homologous desensitization induced by agonist stimulation of transfected beta 2-adrenergic receptors. These observations suggested that ACTH and beta 2-adrenergic agonists homologously desensitized adenylyl cyclase in Y1 cells by a common mechanism. In the present study, parental Y1 cells (Y1DS) and the Y1DR mutant were transfected with the gene encoding the human dopamine D1 receptor and examined for regulation of adenylyl cyclase by dopaminergic agonists. Transformants were isolated from both cell lines and shown to respond to dopamine agonists with increases in adenylyl cyclase activity. Treatment of the Y1DS transformants with ACTH promoted a rapid, homologous desensitization of adenylyl cyclase and had little effect on the responses to dopamine or NaF; treatment of Y1DS with dopaminergic agonists promoted a slower rate of heterologous desensitization that diminished responsiveness of the adenylyl cyclase system to dopamine, ACTH, and NaF. Y1DR cells transfected with the dopamine D1 receptor were resistant to the heterologous desensitization of adenylyl cyclase induced by dopaminergic agonists. These latter observations suggest that the pathways of homologous desensitization and heterologous desensitization converge at a common point in the desensitization pathway defined by the DR mutation in Y1 cells.


[A comparative evaluation of the activity of pyrethroid-based insecticidal preparations].

A comparative analysis of the results of experimental trials of 12 new effective insecticide agents synthesized in the USSR and abroad, containing permethrin, deltamethrin, cypermethrin, cyfenothrin, etophenprox and neopinamine as active ingredients, against flies, mosquitos, fleas, bedbugs and cockroaches was made. The LD50 data in microgrammes per g and millilitres per litre and the LC50 data in grammes per sq m are given. All the tested preparations are allowed to use in medical desinsection practice.


Comparison of glutamine synthetases from brains of genetically epilepsy prone and genetically epilepsy resistant rats.

Since glutamine synthetase (GS) has been proposed as the primary enzyme in the regulation of glutamate metabolism in the central nervous system and since inhibition of the activity of this enzyme in vivo leads to seizures, it has been proposed that an abnormality in the structure or function of this enzyme could be responsible for the induction of seizures in epilepsy prone rats. To test this hypothesis the glutamine synthetases were purified from the brains of both genetically epilepsy prone rats (GEPR) and their progenitors, genetically epilepsy resistant rats (GERR). The enzymes were compared using both SDS-PAGE and isoelectric focusing. The immunoreactivities of equal amounts of protein were determined using the ELISA technique, and the regulation of the glutamine synthetase activities by Mn2+/Mg2+ ratios were compared. The only difference found between the glutamine synthetases from the two strains was a slightly lower specific activity of the enzyme from the epilepsy prone animals.