Left ventricular dysfunction and HLA Bw52 antigen in Takayasu arteritis. Heart disease is the main cause of death in patients with Takayasu arteritis. It has been reported that this disease is closely related to the presence of HLA Bw52 antigen. To assess the correlation between this antigen and left ventricular involvement, we studied 40 patients with Takayasu arteritis, 21 with and 19 without Bw52, using Tl-201 stress myocardial scintigraphy and echocardiography. Those with Bw52 had a significantly higher incidence of abnormal electrocardiographic findings (67% vs 26%; P < 0.05) and of aortic regurgitation (52% vs 11%; P < 0.05). The echocardiographically determined interventricular septal wall thickness plus left ventricular posterior wall thickness (25 +/- 8 vs 17 +/- 3 mm; P < 0.01) and the left ventricular mass (257 +/- 132 vs 142 +/- 51 g; P < 0.01) were significantly increased in the patients with Bw52. Scintigraphically determined perfusion abnormalities were significantly more frequent in those with Bw52 (76% vs 32%; P < 0.05). These observations indicate that patients with Takayasu arteritis and Bw52 antigen have a more severe left ventricular involvement than the patients without that antigen. The left ventricular impairment may account for the poor prognosis of Takayasu patients with Bw52.


Pregnancy in Takayasu arteritis from the view of internal medicine. To evaluate the influence of pregnancy on the morbid condition of Takayasu arteritis, we summarized the clinical data and pregnant courses of 18 patients with Takayasu arteritis and a total of 22 deliveries. We followed C-reactive protein (CRP) scores in 16 of 18 patients (20 of 22 deliveries) to ascertain the inflammatory condition inherent in Takayasu arteritis 1 year prior to, during, and 1 year after pregnancy. We also evaluated digital plethysmograms (pulse amplitude, pulse wave, crest time) to follow the hemodynamical condition of patients before, during, and after pregnancy. CRP scores improved significantly during pregnancy and 1 year after delivery. In the digital plethysmograms, pulse amplitude and wave also exhibited improvement after delivery, but crest time remained unchanged. This indicated that pregnancy is a state favorable to this disease. Some factors, such as the sex hormone progesterone, may induce this condition, but the details are still unknown. In conclusion, inflammatory activity and the hemodynamic state improve with pregnancy in patients with Takayasu arteritis. The physiologic aspects which cause this improvement should be maintained even after pregnancy.


Clinical gynecologic features of pregnancy in Takayasu arteritis. Takayasu arteritis is a non-specific chronic inflammatory vascular disease of unknown etiology. Since the incidence of this disease in the child-bearing years is relatively high, the management of pregnancies with this disease is of great importance in clinical obstetrics. This study is aimed at discussing the maternal management and obstetrical outcome, based on the clinical data obtained from 23 pregnancies of 15 patients treated in our hospital in the past 12 years. Since the disease was in the active state, artificial abortions were conducted in four cases in the 1st trimester of pregnancy. Among the remaining 16 cases, 3, who exhibited neither hypertension nor other complications, vaginally delivered neonates weighing 2,660-3,100 g with Apgar scores of nine after 37 weeks' gestation. C-sections were performed for 13 patients who showed sustained hypertension or/and developed other vascular disorders. Their gestational periods ranged from 34 to 40 weeks and the body weight of the infants varied from 1,425 to 3,024 g. No adverse influence of pregnancy and delivery on Takayasu arteritis was detected in the puerperium of any patients. It is suggested that the state of Takayasu arteritis in early pregnancy and the magnitude of blood pressure elevation in the late gestational period are the most critical and definitive factors in determining the management of pregnancy of a patient with Takayasu arteritis. Cooperative managements by the specialists in obstetrics, internal medicine, and perinatology are required to provide a satisfactory clinical outcome.


Medical treatment of Takayasu arteritis. The guidelines for medical treatment of Takayasu arteritis established in 1987 by the Systemic Vascular Disorders Research Committee, Ministry of Health and Welfare of Japan are presented. The first part of the guidelines concerns treatment with adrenocorticosteroids and the second part concerns other medical treatment. A review of the literature referring to steroid therapy and other medical treatment of Takayasu arteritis is also included.


Takayasu arteritis: follow-up studies for 20 years. We reviewed retrospectively 126 (5 male, 121 female) patients suffering from Takayasu arteritis who had been treated in our clinics from 1971 to 1990. The patients' ages ranged from 19 to 80 yrs old (1990) with a mean age of 48.7 +/- 11.8 years. HLA typing analysis in 98 patients revealed that 45 patients (47%) were confirmed as carrying the Bw52 antigen, a high result that is statistically significant as compared with that in healthy Japanese. Arteriograms (performed in 75 patients) revealed that 28 patients (37%) were affected in the aorta and its main branches by this disease (type IV by Nasu's classification) and 23 patients (31%) were affected only in the main branches (type I). The C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) improved significantly from 2.55 +/- 0.28(+) and 57.0 +/- 5.69 mm/hr to 0.53 +/- 0.12(+) and 31.2 +/- 3.45 mm/hr, respectively after treatment including steroid and antiplatelet therapy (P < 0.01). Patients with Bw52 exhibited more severe inflammatory conditions than those without Bw52. Lung scintillations performed in 81 patients showed pulmonary arterial lesions in 50 patients (62%). Echocardiograms revealed aortic regurgitation (AR) in 44 patients (35%), with a significant difference noted between the Bw52 positive group and the Bw52 negative group [29/40 (73%) versus 11/47 (23%), respectively, P < 0.001]. Patients with Bw52 were prescribed higher doses of steroids (P < 0.05) for longer periods (P < 0.01) than those without Bw52.(ABSTRACT TRUNCATED AT 250 WORDS)


Percutaneous transluminal angioplasty in Takayasu arteritis. Percutaneous transluminal balloon angioplasty (PTBA) was performed in 87 patients for 111 stenotic lesions due to Takayasu arteritis. Of the lesions attempted for dilatation, 35 were in the aorta, 64 in renal arteries, 9 in subclavian, and 3 in common iliac arteries. The stenosis of aorta could be successfully dilated in 33 of 35 (94.3%) patients with fall in peak systolic pressure gradient (PSG) from 77.7 +/- 28.4 mmHg to 26.4 +/- 20.6 mmHg (P < 0.001) and increase in luminal diameter from 4.7 +/- 2.4 mm to 10.1 +/- 4.1 mm (P < 0.001). On hemodynamic and angiographic restudy in 20 patients at 3-24 months (mean 7.7 +/- 4.1 months) further fall in PSG (> or = 15 mmHg) was observed in 7 patients, no significant change in 12 patients and restenosis with increase in PSG in one patient which could be successfully redilated. Late restudy at 36-60 months (mean 43 +/- 9.4) in six patients showed continued relief of stenosis (mean PSG 8.8 +/- 7.8 mmHg). Of the 64 stenotic lesions of the renal arteries, 58 (90.6%) could be successfully dilated with decrease in stenosis from 89.1 +/- 10.1% to 29.9 +/- 14.9% (P < 0.001). Follow-up intra-arterial digital subtraction angiography in 25 patients at a mean follow-up period of 13.1 months (range 3-29 months) showed restenosis in 5/36 (13.9%) lesions which could be successfully redilated. Angioplasty was also successful in dilating 8/9 (88.9%) subclavian and all 3 common iliac artery stenosis.(ABSTRACT TRUNCATED AT 250 WORDS)


Surgical treatment of Takayasu arteritis. The role of surgical therapy for Takayasu arteritis remains controversial. From 1973-1991, 23 patients with Takayasu arteritis have been treated at the University of Southern California. Twelve patients have required 17 arterial reconstructions for symptomatic complications of arterial disease refractory to medical therapy. Indications for operation have included renovascular hypertension (7), extremity ischemia (5), cerebrovascular insufficiency (2), dilated ascended aorta with aortic insufficiency (1), thoracic aortic aneurysm (1), and abdominal aortic aneurysm (1). Long-term clinical follow-up has demonstrated uniform symptomatic improvement. Fifteen of seventeen arterial reconstructions are still patent. Surgical treatment of symptomatic Takayasu arteritis is highly effective. Excellent long-term graft patency can be expected following arterial reconstruction.


Surgical treatment of Takayasu arteritis. From 1959 to 1991, 93 patients underwent vascular reconstruction for Takayasu arteritis at our institution. The details of the cases were as follows: 16 were of type I (brachiocephalic ischemia), 48 type II (hypertension), 13 type III (extensive lesions with cerebral ischemia and hypertension), and 16 type IV (aneurysms). Carotid reconstruction, repair of atypical aortic coarctation, renovascular reconstruction, and aneurysm repair were performed independently or in combination. Nine operative deaths occurred, 8 cases of which were operated before 1970. The most serious of the delayed complications was suture line aneurysm formation, which was encountered in ten cases. The aneurysms were often found long after the operation, some of them developing even after more than 20 years. Takayasu arteritis is characterized by extensive inflammation and destruction of the medial elastic fibers and long term postoperative observation is mandatory to improve the late survival rate.


Pathological features of the pulmonary artery in Takayasu arteritis. Little attention has been paid to the pathological features of the pulmonary artery in Takayasu arteritis. Autopsy specimens of 6 cases of this disease were studied. Lesions were found in the aortic arch and its brachiocephalic branches in all cases and in both the aortic arch and thoracoabdominal aorta in 5 cases. The pathohistologic characters of the pulmonary artery were very similar to those of the systemic artery. Stenosis-recanalization, so-called blood vessels-in-blood vessels, of the pulmonary elastic arteries were found in four cases. These lesions were not observed in the systemic arteries, and most of the newly formed channels in them seemed to be branches of bronchial arteries. Luminal obstruction of pulmonary muscular arteries was observed in 4 cases, cellular arteritis of muscular arteries in 2 cases, and angiomatoid dilatation of small blood vessels in 2 cases. Thus in this study we found peculiar stenosis-recanalization lesions of the pulmonary elastic arteries, and also showed that the pulmonary elastic and muscular arteries are frequently involved in Takayasu arteritis. These findings suggest that pulmonary hypertension could influence morbidity and long-term mortality in this disease.


Surgical treatment of cardiac involvement in Takayasu arteritis. Cardiac involvement is a serious disorder in Takayasu arteritis. Surgical treatment of aortic root and coronary artery lesions due to Takayasu arteritis has many potential difficulties due to its inflammatory nature. We operated on 15 patients with cardiac involvement stemming from Takayasu arteritis. These patients are classified into 3 groups depending on the clinical diagnosis and surgical procedures employed: coronary artery involvement alone--coronary artery bypass grafting (CABG), three patients (group A), aortic regurgitation with intact coronary artery--Aortic valve replacement or modified Bentall's operation, eight patients (group B), and aortic regurgitation with coronary artery involvement g aortic valve replacement or modified Bentall's operation with CABG, (4 patients) (group C). CABG was performed using saphenous vein graft. For aortic valve replacement or replacement of both the aortic valve and ascending aorta, a prosthetic valve or composite graft with Teflon felt flange was fabricated during surgery and treated with fibrin glue before insertion. A double fixation method with reinforcement by a Teflon felt strip was employed for proximal anastomosis of the flanged prosthesis. A button-shaped coronary ostium was directly anastomosed to the composite graft. There was no operative or hospital mortality. One patient died of brain abscess at 6 months after surgery and another patient with CABG required a second operation due to graft failure. Steroid therapy is recommended in cases diagnosed as being in an active stage until the inflammatory signs disappear.


Coronary artery lesions in Takayasu arteritis: pathological considerations. This communication reviews the clinical and pathological features of coronary artery lesions in Takayasu arteritis. The incidence of coronary artery involvement has been reported to be 9% to 10%, and is observed mainly in autopsy cases because coronary artery disease is usually not evident until the occurrence of angina pectoris or myocardial infarction, or after the onset of congestive heart failure. On the basis of pathological features, the following three types of coronary artery lesions can be distinguished: type 1, stenosis or occlusion of the coronary ostia and the proximal segments of the coronary arteries; type 2, diffuse or focal coronary arteritis, which may extend diffusely to all epicardial branches or may involve focal segments, so-called skip lesions; and type 3, coronary aneurysm. Most of the coronary artery lesions in Takayasu arteritis are of type 1. Narrowing of the coronary arteries is mainly due to the extension of the inflammatory processes of proliferation of the intima and contraction of the fibrotic media and adventitia from the ascending aorta. In some cases, coronary stenosis may be caused by coronary arteritis as skip lesions in Takayasu arteritis, but even in these cases the lesions have been reported to affect mainly the proximal segments of the coronary arteries. Diffuse lesions of the coronary artery and coronary artery aneurysm seem to be very rare in Takayasu arteritis. Other causes of coronary ostial stenosis, coronary arteritis and coronary artery aneurysm are also discussed.


Takayasu arteritis in China: a report of 530 cases. A total of 530 patients with Takayasu arteritis were studied. Among 346 patients who were diagnosed by aortography, the female to male ratio was 2.9 to 1, and the age of onset ranged from 5 to 45 years. Three hundred and eighteen (60%) patients with secondary hypertension, including renovascular hypertension in 281, and 197 (37.2%) with pulseless extremities were found in the series. All the patients were treated with medical or surgical procedures. Surgical treatment is preferred if clinical status of the patient permits. The patients were followed for 1-29 years (average 7.8 years). There were 55 deaths (10.4%) in this series. Cerebral hemorrhage was found as a common cause of death. Five-year and ten-year survival rates were 93.1% and 91.1%, respectively.


Takayasu arteritis in India.

Takayasu arteritis in India. Takayasu arteritis is the commonest cause of renovascular hypertension in India. The clinical and radiological features, complications and course of 83 patients (51 females, 32 males) seen during the period from 1972-1990 are described in this study. The age of the patients ranged from 5 to 53 years with the mean +/- SD of 26.9 +/- 9.7. Hypertension (n = 50) and the related symptom of headache (n = 40), dyspnea (n = 24), and giddiness (n = 20) were common at presentation. Twelve patients were in congestive cardiac failure. The symptoms of activity with fever and arthralgia were present in only 16% contrary to reports from Japan and Mexico. Abnormal arterial pulses and bruit over abdominal (37%) or extra abdominal great arteries (25%) were useful clinical clues to suspect Takayasu arteritis. Rapid sequence intravenous urography was a sensitive screening procedure and predicted correctly the presence of renovascular disease in 80% of the patients. The diagnosis was confirmed on aortography in 72. In the rest, the clinical features and autopsy findings confirmed the same. The four patterns of the disease based on the anatomical extent of involvement were recognised. These were: type I (n = 8) with involvement of aortic arch and its branches, type II (n = 25) descending thoracic and abdominal aorta type III (n = 46) combination of I and II and type IV (n = 4) pulmonary artery in addition to any of the above.(ABSTRACT TRUNCATED AT 250 WORDS)


Takayasu arteritis in Japan.

Takayasu arteritis in Japan. Takayasu arteritis is a disease on which many investigations have been conducted to determine its causes, clinical features and treatment, since 1908 when it was reported by Dr. Takayasu. The first nationwide epidemiological survey on Takayasu arteritis was conducted over 3 years from 1973 through 1975 by a research group on "aortitis syndrome", a disease specified by the Ministry of Health and Welfare. Another nationwide survey on this disease over a period of 3 years was carried out from 1982 through 1984 by a research group focusing on "systemic vascular lesion", another disease specified by the Ministry of Health and Welfare. The present study reports on the combined results of these two nationwide epidemiological surveys.


Takayasu arteritis in Korea: clinical and angiographic features. Clinical and angiographic features of Takayasu arteritis were investigated in 129 Korean patients. This disease affects females more frequently than males, in a ratio of 6.6 to 1. Of the total number of patients, 51 were in the third decade, 27 in the fourth decade, and 23 in the second decade. Common clinical symptoms were headache (60%), exertional dyspnea (42%), dizziness (36%), and malaise or weakness (34%). Takayasu arteritis affected the abdominal aorta (46%) and descending thoracic aorta (37%) more frequently than the ascending aorta (1%) and aortic arch (2%) According to Ueno's classification based on aortographic findings, the 129 patients were divided into type I (37), type II (25), and type III (67). Among the 48 patients who had coronary angiography, 11 (23%) showed coronary arterial involvement. Because the clinical features are determined by the extent and severity of the specific artery involved in the occlusive phase of the disease, total aortography including coronary angiography is very important in the initial evaluation of Takayasu arteritis.


An overview on Takayasu arteritis.

An overview on Takayasu arteritis. Takayasu arteritis is a non-specific inflammatory disease of unknown etiology. It was first recognized as having a peculiar wreath-like arteriovenous anastomosis around the papillae of the retina by a Japanese ophthalmologist, Dr. M. Takayasu in 1908. A Japanese research committee reported more than 5,000 cases. For a supplement issue on Takayasu arteritis, this brief overview article has been written as an introduction to the disease.


Takayasu arteritis in Thailand.

Takayasu arteritis in Thailand. Takayasu arteritis is a disease of world-wide distribution. Geographic difference in sex incidence, anatomical distribution, and the type of lesion is observed. Hypertension is very common in the present series, as well as in combination with absent or deficit of peripheral pulses. These symptoms correlate well with arteriographic findings of brachiocephalic and renal artery obstructive lesion. While aneurysm and stenotic lesions have a predilection in the abdominal aorta, stenotic lesion of the thoracic aorta occurs more commonly than aneurysm. However, aneurysm of the aorta as well as of the brachiocephalic arteries is seen more frequently than in the reports, of others. The presence of "funnel-shape" resulting from renal artery obstructive lesion and dilatation or aneurysm of contiguous aorta was characteristic of Takayasu arteritis in our series. The material presented in this report reflects not only geographic variation but also the severe form of this disease. Total aortography, coronary arteriography and pulmonary arteriography, are of value not only for the diagnosis of Takayasu arteritis but also for demonstration of anatomical distribution, severity and type of lesion.


Hereditary factors of Takayasu arteritis. Takayasu arteritis is a chronic vasculitis characterized by a clinical pulseless condition and is predominant in young female patients. Its loci is found mainly in Asian countries, and its etiology is still unknown. Our experiences of cases of twin sisters with Takayasu arteritis led us to suppose that hereditary factors participate in the pathophysiology of this disease. Population and family-incidence studies employing HLA analysis in Japan have focused on an complotype Aw24-DW52-C4A2-C4BQ0-Dw12 which was in disequilibrium with Takayasu arteritis. Clinical features and clinical courses were found to be intimately related to this complotype. Recent studies on HLA typing in other countries have also suggested the important roles of hereditary factors in this morbid condition and international collaborative studies on these hereditary factors are now under way.


HLA-linked susceptibility and resistance to Takayasu arteritis. To investigate genetic factors involved in the pathogenesis of Takayasu arteritis, patients in the Japanese population were examined for HLA-A, -B, and -C alleles by serological typing and for HLA-DR, DQ, and DP alleles by DNA typing using polymerase chain reaction (PCR)/sequence-specific oligonucleotide probe (SSOP) analysis. The frequencies of HLA-Bw52, DRB1*1502, DRB5*0102, DQA1*0103, DQB1*0601, and DPB1*0901 alleles were significantly increased and the frequencies of HLA-Bw54, DRB1*0405, DRB4*0101, DQA1*0301, and DQB1*0401 alleles were significantly decreased. Strong linkage disequilibria among the increased alleles and among the decreased alleles were evident in the Japanese population. Therefore, the haplotype of HLA-B252-DRB1*1502-DRB5*0102-DQA1*0103-DQB1++ +*0601-DPA1*02-DPB1*0901 may confer susceptibility to Takayasu arteritis while another haplotype of HLA-Bw54-DRB1*0405-DRB4*0101-DQA1*0301-DQB1++ +*0401 may confer resistance to the disease. These observations clearly indicate that HLA-linked gene(s) are involved in the development of Takayasu arteritis.


HLA typing of Takayasu arteritis in Korea. Takayasu arteritis occurs with a strong predilection for women and particular geographic areas, and as related to the etiology of the disease, association of HLA antigens has been suggested. In the present study, the authors investigated the association of Takayasu arteritis with class I and class II HLA antigens in 59 Korean patients with this disease. Increased frequencies of HLA-Bw52 (chi 2 6.213, P < 0.02), Cw6 (chi 2 4.132, P < 0.05), DR7 (chi 2 4.506, P < 0.04), and DQw2 (chi 2 7.327, P < 0.01) were observed in the patient group as compared to the control group of healthy Koreans. In the Korean population, 2 risk factors in the HLA system for developing this disease appear to be (1) Bw52 and (2) DR7 and a probable haplotype of Cw6, B13, DR7, DQw2. Previous studies of the Japanese population revealed association of Bw52 and class II HLA antigens (DR2, Dw12), which are in linkage disequilibrium with Bw52. It is of interest that in the Korean population, class II antigens (DR7, DQw2), which are not linked to BW52, are associated with the disease. This finding suggests that the disease susceptibility gene of Takayasu arteritis is located between the HLA-B locus and HLA-DR, DQ loci.


Immunopathogenesis of Takayasu arteritis. Takayasu arteritis is a common cause of renovascular hypertension in India. Sensitization to infective agents, particularly mycobacterium tuberculosis or autoimmune disturbances have been incriminated in its pathogenesis. Twenty patients of Takayasu arteritis along with groups of normal individuals, patients of essential hypertension, autoimmune disorders, tuberculosis, and healthy tuberculin reactors were studied. Besides detailed immunological profiles including LE cell phenomenon, serum complement C3 levels, antibodies to single (SS) and double stranded (DS) DNA, rheumatoid factor, lymphocyte subsets, blast transformation to antigens including, phytohemagglutinin, PPD, pokeweed, and purified human aortal antigen (PHAA) were examined. Soluble protein from human aorta was fractionated into 9 peaks by DEASE-52 and Sephadex G-75 chromatography, and 25 micrograms of major protein fraction-II was used for blast transformation study. Blast transformation by PHAA was higher in Takayasu arteritis as compared to all other groups (P < 0.05). Blast transformation to PPD showed wide variation in all the groups, and was significantly higher only in tuberculin reactors as compared to controls. These observations support aortal sensitization to PHAA playing a role in pathogenesis of Takayasu arteritis and do not relate tuberculosis to Takayasu arteritis, at least immunologically. In addition, the ratio of CD-4 positive to CD-8 positive lymphocytes changing in favor of the former and the concomitant increase in B lymphocytes favor the presence of autoimmune disturbances in Takayasu arteritis.


Takayasu arteritis in Korean children: clinical report of seventy cases. Seventy cases of Takayasu arteritis in Korean children are reported. There were 57 females and 13 males (male-to-female ratio; 1:4.4). The youngest patient was a 3-year-old female. Family history was positive in one patient. The most common chief complaints on admission were dyspnea, headache, palpitation, and edema which were due to hypertension and congestive heart failure. Hypertension was seen in 65 out of 70 patients (92.8%). The abdominal aorta, thoracic aorta, and renal arteries were the most commonly involved sites in these children. Two patients had nephrotic syndrome. The frequency of positive tuberculin reaction was much higher in children with Takayasu arteritis compared with the general population, and the intensity of the reaction was also stronger. The majority of the patients required immediate medical treatment to control congestive heart failure due to hypertension at initial presentation. When ESR was elevated, corticosteroid was administered. Surgical treatment showed good results in six out of ten cases. Percutaneous intraluminal angioplasty was effective for lowering the blood pressure in six out of nine cases. In three cases, restenosis occurred and angioplasty was repeated in two cases.


Modulation of P-glycoprotein-mediated drug transport by alterations in lipid fluidity of rat liver canalicular membrane vesicles. P-glycoprotein (P-gp) is believed to function as an ATP-dependent efflux pump for natural product anti-cancer drugs in multidrug-resistant (MDR) tumor cells and in certain normal tissues. P-gp has been localized to the apical plasma membrane of the bile canaliculus where it has been shown to transport [3H]daunomycin. In this study, we investigated whether alterations in membrane lipid fluidity of canalicular membrane vesicles (CMV) could modulate the P-gp-mediated accumulation of [3H]daunomycin and [3H]vinblastine. Accumulation of both cytotoxic agents was stimulated by ATP, exhibited temperature dependence and osmotic sensitivity, and followed Michaelis-Menten kinetics. Alterations in CMV lipid fluidity were induced by the known fluidizers, 2-(2-methoxyethoxy)ethyl 8-(cis-2-n-octylcyclopropyl)octanoate (A2C) and benzyl alcohol, and were assessed by fluorescence polarization techniques using the fluorescent probe, 1,6-diphenyl-1,3,5-hexatriene (DPH). Both A2C (2.5-5.0 microM) and benzyl alcohol (10-20 mM) produced a dose-dependent increase in CMV lipid fluidity. Moreover, both fluidizers, at the above doses, significantly inhibited (p < 0.05) the ATP-dependent accumulation of [3H]daunomycin. [3H]Vinblastine accumulation was also inhibited by A2C (p < 0.05). Lower doses of A2C (0.6 microM) and benzyl alcohol (1 mM) failed to influence either lipid fluidity or P-gp-mediated drug accumulation. Kinetic analysis revealed that A2C (5.0 microM) noncompetitively inhibited [3H]daunomycin accumulation and uncompetitively inhibited [3H]vinblastine accumulation with apparent Ki values of approximately 1.5 and approximately 1.2 microM, respectively. Verapamil competitively inhibited P-gp-mediated accumulation of [3H]daunomycin but failed to alter the fluidity of CMV. Taken together, the present results demonstrate that while increases in membrane fluidity of CMV are not necessarily required to inhibit P-gp-mediated drug accumulation, they can inhibit these processes, at least in CMV. Alterations in the physical state of CMV, therefore, appear to be at least one important modulator of P-gp function.


Identification of glutamic acid 105 at the active site of Bacillus amyloliquefaciens 1,3-1,4-beta-D-glucan 4-glucanohydrolase using epoxide-based inhibitors. Bacillus amyloliquefaciens 1,3-1,4-beta-D-glucan 4-glucanohydrolase (EC 3.2.1.73) was modified by the mechanism-based, affinity-labeling reagent [14C](3,4)-epoxybutyl beta-D-cellobioside. Following partial inactivation a completely inactivated enzyme preparation containing 1.1 mol of covalently bound inhibitor/mol of protein was obtained by chromatography on a cellulosic matrix. The inactivated enzyme was digested with endoproteinase Glu-C and radioactive peptides purified by reversed-phase high performance liquid chromatography (HPLC). The affinity label was esterified exclusively to the gamma-carboxylate of Glu105 in the sequence Gly-Thr-Pro-Trp-Asp-Glu-Ile-Asp-Ile-Glu109. The sequence motif Glu-(Ile/Leu)-Asp-Ile is found in many glucanases and xylanases and may therefore serve to identify the catalytic nucleophile in beta-glycanases, which otherwise exhibit a low degree of sequence identity. The esterification of Glu105 by the affinity label abolished endoproteinase Glu-C-mediated hydrolysis of the Glu-Ile106 peptide bond. Identification of phenylthiohydantoin-Glu105 during automated sequence analysis was not possible unless the affinity label was liberated by prior base hydrolysis. These observations formed the basis for the development of a highly sensitive approach for the identification of catalytic carboxylates in polysaccharide hydrolases employing non-radioactive inhibitors, comparative HPLC mapping, electrospray mass spectrometry, and Edman degradation.


Identification of residues in the first cytoplasmic loop of P-glycoprotein involved in the function of chimeric human MDR1-MDR2 transporters. The human MDR1 gene encodes the multidrug transporter (P-glycoprotein), a multidrug efflux pump. The highly homologous MDR2 gene product does not appear to be a functional multidrug pump. We have constructed a chimeric protein in which the first intracytoplasmic loop and the third and fourth transmembrane domains of the MDR1 protein were replaced by the analogous region of MDR2. Substitution of the MDR2 sequences encompassing amino acid residues 140 to 229 resulted in 17 amino acid changes, 10 in the intracytoplasmic loop (amino acids 141-188) and 7 in the transmembrane regions. This chimeric protein was expressed on the surface of NIH 3T3 cells where it bound [3H]azidopine but did not confer drug resistance. When only 4 residues, 165, 166, 168, and 169, were changed back to MDR1 amino acids, a functional drug transporter was recovered. When residues 165, 166, 168, and 169 from MDR2 were substituted into a functional MDR1 cDNA, the resulting construction was not able to confer drug resistance. These results indicate that the major functional differences between MDR1 and MDR2 in this region of P-glycoprotein reside in a small segment of the first intracytoplasmic loop. We also independently analyzed the effect of replacing Asn183 of MDR1 with Ser which occurs in MDR2. Substitution of Ser at position 183 in combination with Val at position 185 in P-glycoprotein resulted in a relative increase in resistance to actinomycin D, vinblastine, and doxorubicin in transfected NIH 3T3 cells. These results emphasize the importance of the first intracytoplasmic loop in P-glycoprotein in determining function and relative drug specificity of the transporter.


An analysis of response prevention and flooding procedures in the treatment of adolescent obsessive compulsive disorder. Flooding and response prevention have been widely used in the treatment of adult obsessive compulsive disorder but have been overlooked in favor of less restrictive procedures when treating children. The present case investigates the utility of these procedures in an adolescent with severe compulsive handwashing. Treatments were introduced hierarchically to minimize subject distress; graded exposure decreased the frequency of handwashing, but flooding was required to eliminate the compulsion. Guidelines for the ethical use of flooding and exposure therapies with children are offered.


Patterns of glutamate, glycine, and GABA immunolabeling in four synaptic terminal classes in the lateral superior olive of the guinea pig. The goal of this study was to correlate synaptic ultrastructure with transmitter specificity and function in the lateral superior olive (LSO), a nucleus that is thought to play a major role in sound localization. This was accomplished by means of postembedding immunogold immunocytochemistry. Four classes of synaptic terminals were identified in the LSO. They were distinguishable from one another both morphologically and on the basis of their different patterns of immunolabeling for glutamate, glycine, and gamma-aminobutyric acid (GABA). The highest level of glutamate immunoreactivity was found in terminals that contained round vesicles (R) and formed synaptic contacts with asymmetric synaptic junctions. Round-vesicle terminals predominated on small caliber dendrites by a ratio of at least 2:1 over the other classes combined. The thinnest dendrites were typically contacted by R terminals only. The ratio of R terminals to the other types decreased as the caliber of the dendritic profiles they apposed increased so that on the soma, R terminals were outnumbered by at least 2:1 by the other types. Terminals containing flattened vesicles (F) exhibited intense immunoreactivity for both glycine and glutamate, although the glutamate immunolabeling was not as high as that in the R terminals. Flattened-vesicle terminals formed symmetric synaptic contacts with their targets and their distribution was the reverse of that described for R terminals; i.e., they were most abundant on LSO perikarya and fewest on small caliber dendrites. Two terminal types, both containing pleomorphic vesicles and forming symmetric synaptic junctions, were found in far fewer numbers. One group contained large pleomorphic vesicles (LP) and was immunoreactive for both glycine and GABA. The other group contained small pleomorphic vesicles (SP) along with a few dense-core vesicles and labeled for GABA only. The LP terminals were preferentially distributed on somata and large-caliber dendrites, while the SP terminals most often contacted smaller dendrites. Previous work suggests that a large percentage of the R terminals arise from spherical cells in the ipsilateral cochlear nucleus and are excitatory in action. This pathway may use glutamate as a transmitter. Many of the F terminals are thought to originate from the ipsilateral medial nucleus of the trapezoid body and appear to be the inhibitory (glycinergic) terminals from a pathway that originates from the contralateral ear. The origins and functions of LP and SP terminals are unknown, but a few possibilities are discussed along with the significance of cocontainment of neuroactive substances in specific terminal types.


Growth properties of larval and adult locust neurons in primary cell culture. We developed a cell culture system for thoracic neurons of fifth instar or adult locusts (Locusta migratoria) in order to obtain maximum visualization of cellular morphology and direct access to the neurons for electrophysiological analysis. The dissociated neurons regenerated new neurites in a serum-free defined culture medium, in which they remained viable for up to 3 weeks. Viability of the cells was confirmed by intracellular recordings demonstrating active membrane properties and action potentials. While the morphology of the cultured neurons is distinct from their in vivo counterparts, they retained some cellular surface properties and markers related to transmitter metabolism. Two factors influencing cellular morphology in vitro were identified in Locusta: 1) the presence of a primary neurite stump, and 2) membrane contacts between cells. Dissociated neurons of the locust species Schistocerca gregaria grown in a hemolymph-enriched medium showed a marked reduction in branching patterns and a tenfold increase in neurite length compared to neurons growing in a medium without hemolymph. This culture system could prove useful for identifying the action of hemolymph-derived growth factors.


Comorbidity of anxiety and depression in youth: treatment implications. Reviews the empirical literature on the comorbidity of anxiety and depressive disorders in youth, emphasizing prevalence of comorbidity, difficulties in assessment and measurement, familial factors, and developmental differences. The nature of anxiety and of depression in youth is examined (e.g., differentiating cognitive deficiencies from cognitive distortions), and treatment recommendations are presented from a cognitive-behavioral framework. Components of the treatment include affective education, enactive programming, addressing reinforcement difficulties, correcting cognitive distortions, and enhancing problem-solving skills. Peer and familial factors are discussed. Successful treatment of co-morbid children relies on a flexible application of these strategies with consideration of the developmental level and particular symptom constellation of the individual child.


Attention-deficit hyperactivity and conduct disorder: comorbidity and implications for treatment. The distinguishing and overlapping features of attention-deficit hyperactivity disorder (ADHD) and conduct disorder (CD) are discussed. Conclusions regarding comorbidity, treatment efficacy, and long-term outcome can be influenced by several factors, including diagnostic procedures and sample characteristics. The need to distinguish between referred and non-referred samples is particularly crucial when considering treatment and comorbidity issues. The efficacy of psychosocial and pharmacological treatments in ADHD and CD children is reviewed as are the few studies of psychostimulant medication in co-morbid youngsters. Suggestions regarding treatment planning and recommendations for treatment and research are described.


The effects of phenindamine tartrate on sleepiness and psychomotor performance. Phenindamine, an H1-receptor antagonist that was developed almost 50 years ago, has been associated with both drowsiness and insomnia. Since its central nervous system profile has not been well characterized, we used a series of psychomotor tests to conduct two studies. In the first, 12 subjects received single oral doses of phenindamine (25 mg), diphenhydramine (50 mg), terfenadine (60 mg), or placebo in a four-way crossover study. Psychomotor tests included choice reaction time (CRT), tracking, and hand steadiness (HS). In the second trial, 15 subjects received single oral doses of phenindamine (25 mg), pseudoephedrine (60 mg), phenindamine and pseudoephedrine, diphenhydramine (50 mg), or placebo in a five-way crossover study. Psychomotor tests included CRT, HS, and a task that divided attention between tracking and reaction time. Introspective drowsiness was measured in both trials with use of a visual analog scale (VAS) and the Stanford Sleepiness Scale (SSS). All assessments were made before and 1, 3, and 5 hours after drug administration. In the first trial, diphenhydramine produced significant impairment relative to placebo (p < 0.05) in CRT, tracking, and HS tasks and higher SSS and VAS scores, with peak effect noted at 3 hours. Phenindamine did not significantly differ from placebo or terfenadine. In the second trial, diphenhydramine produced significant impairment relative to placebo (p < 0.05) in CRT, divided attention, HS, and VAS, and SSS, also peaking at 3 hours. Stanford Sleepiness Scale scores after phenindamine were greater than placebo at 3 hours (p < 0.05) but significantly less than diphenhydramine (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Distribution of sympathetic neuroeffector junctions in the juxtaglomerular region of the rabbit kidney. Two structurally distinct types of sympathetic axon (Type I and Type II) have recently been identified in the renal cortex of the rat and the rabbit. This study describes the distribution and density of the neuroeffector junctions made by these two types of axon on the different tissues from the juxtaglomerular region of the rabbit renal cortex. Immunohistochemical studies showed that tyrosine hydroxylase-positive axons were located only in regions adjacent to the arteries and arterioles in the renal cortex. Ultrastructural studies of the juxtaglomerular region indicated that both types of axon formed junctions on vascular smooth muscle cells, epithelial cells of proximal tubules and renin-secreting granular epithelioid cells. The density of neuromuscular junctions (18 x 10(3)/mm2 of vessel surface) was more than twice as high on the afferent arteriole as on the efferent arteriole or proximal tubules immediately adjacent to the glomerular arterioles (both about 6 x 10(3)/mm2). The junction density on granular epithelioid cells was much lower (about 2 x 10(3)/mm2) and were rarely observed on the distal tubule. Afferent arterioles preferentially received junctions from Type I axons at a relatively high density (14.2 x 10(3)/mm2) whereas junctions formed by Type II axons were less selectively distributed and occurred at lower densities on all other tissues (range, 1-6.3 x 10(3)/mm2). Presynaptic membrane specialisations were identified only at junctions on arterioles and granular epithelioid cells and occurred more frequently at Type I than at Type II junctions. The data suggest that the predominant effect of the sympathetic innervation in the juxtaglomerular region of the renal cortex is on the afferent arteriole and that the two axon types within the kidney may have different functions.


IL-12 augments antigen-dependent proliferation of activated T lymphocytes. Ag-dependent T cell activation requires multiple transmembrane signals including activation of Ag-specific T cell receptor in combination with signals delivered through cytokine receptors. IL-12 is a heterodimeric cytokine involved in the regulation of NK and T lymphocyte responses. In examining the role of IL-12 in T cell activation, we found a direct relationship between Ag stimulation and IL-12-induced proliferation. Unlike IL-2, which induced proliferation of CTL either in the presence or absence of a CD3/TCR co-signal, IL-12 mediated proliferation of CTL only when the cells were recently co-stimulated with alloantigen or solid-phase anti-CD3 antibody. After culture in the absence of alloantigen or anti-CD3 for 7 to 14 days, these CTL lost the ability to proliferate to IL-12 alone. Under these conditions, however, IL-12 synergized with low-dose IL-2 to induce CTL proliferation. Restimulation with alloantigen or solid-phase anti-CD3 restored the ability of the CTL to proliferate to IL-12 alone. Not all Ag signals resulted in IL-2 independent proliferation to IL-12. For example, CTL with specificity for influenza matrix peptide proliferated best when co-cultured with peptide Ag presented on self MHC and a combination of IL-2 and IL-12. This evidence suggests that IL-12 may be useful in expanding an Ag-specific T cell population, as the culture of CTL with IL-12 and low-dose IL-2 leads to proliferation only in response to an Ag co-signal.


Costimulation of T cell receptor/CD3-mediated activation of resting human CD4+ T cells by leukocyte function-associated antigen-1 ligand intercellular cell adhesion molecule-1 involves prolonged inositol phospholipid hydrolysis and sustained increase of intracellular Ca2+ levels. Activation of resting human CD4+ T cells mediated by mAb ligation of the TCR/CD3 complex requires costimulatory signals to result in proliferation; these can be provided by intercellular cell adhesion molecule-1 (ICAM-1, CD54) a natural ligand of leukocyte function-associated Ag-1 (LFA-1, CD11a/CD18). We analyzed early signaling events involved in T cell activation to determine the contribution by the LFA-1/ICAM-1 interaction. We studied in detail the hydrolysis of phosphatidylinositol(4,5)bisphosphate and intracellular levels of free Ca2+ during stimulation with beads coated with the CD3 mAb OKT3 alone or in combination with purified ICAM-1 protein. Our investigations show no response to LFA-1/ICAM-1 alone, but that costimulation by LFA-1/CAM-1 interaction induces prolonged inositol phospholipid hydrolysis (up to 4 h), resulting in generation of both inositol(1,4,5)phosphate3 and inositol(1,3,4,5)phosphate4 and their derivatives. Based on studies with cycloheximide, this costimulatory effect of prolonged inositol phospholipid hydrolysis appears dependent in part on de novo protein synthesis. A sustained increase in intracellular levels of free Ca2+ level is also observed after LFA-1/ICAM-1 costimulation, which is at least partly dependent on extracellular sources of Ca2+. Kinetic studies indicate that costimulation requires a minimal period of 4 h of LFA-1/ICAM-1 interaction to provide maximal costimulation for OKT3-mediated T cell proliferation. Thus, the necessary costimulation required for OKT3-mediated proliferation in this model system may be provided by an extended LFA-1/ICAM-1 interaction that in combination with OKT3 mAb leads to signal-transducing events, resulting in prolonged phospholipase C activation and phosphatidylinositol(4,5)bisphosphate hydrolysis, and a sustained increase in intracellular levels of free Ca2+.


Lipopolysaccharide increases glucocorticoid receptor expression in murine macrophages. A possible mechanism for glucocorticoid-mediated suppression of endotoxicity. In a previous study we demonstrated that IFN-gamma induced an increase in the number of glucocorticoid receptors (GR) in murine macrophages. To examine further the environmental signals involved in regulation of macrophage GR availability, we asked whether another classical macrophage-activating factor, LPS, would induce an increase in GR number in the macrophage cell line, RAW 264.7, and in primary macrophages from C3H mice. We report that treatment of RAW 264.7 cells and peritoneal exudate macrophages from C3H/OuJ mice with protein-free, phenol water-extracted LPS (PW-LPS) induced an increase in the number of GR. A significant increase in GR number was observed as early as 4 h after PW-LPS treatment, was maximal at 12 h, and remained heightened through 48 h. Optimal induction of the GR by PW-LPS was observed when murine macrophages were treated with 10 ng/ml of PW-LPS. The LPS-induced increase in macrophage GR number could be inhibited by polymyxin B. Macrophages obtained from the LPS hyporesponsive C3H/HeJ strain did not respond to PW-LPS, but did respond to protein-rich, butanol-extracted LPS with a modest increase in GR number after treatment with 2 micrograms/ml. Moreover, taxol, an antineoplastic agent with LPS mimetic activity, also increased GR number in murine macrophages. These results suggest that LPS is not only an important macrophage-activating signal, but may also be important in sensitizing the cell for negative regulatory events such as feedback inhibition by glucocorticoids.


Differential expression of VLA-alpha 4 and VLA-beta 1 discriminates multiple subsets of CD4+CD45R0+ "memory" T cells. Given the importance of adhesion in T cell development, we have undertaken systematic flow cytometric analysis of CD4 T cells to determine relationships between the developmentally regulated marker CD45R0 and adhesion receptors (five VLA integrin chains). The most important findings are that: 1) expression of alpha 3, alpha 5, and alpha 6 are closely coregulated with beta 1 on CD4 cells, while regulation of VLA-alpha 4 is quite discordant. 2) CD45R0- cells, generally understood to be naive cells, have low homogeneous expression of VLA-alpha 3, VLA-alpha 4, VLA-alpha 5, VLA-alpha 6, and beta 1 integrin chains; studies of cord blood CD4 cells confirm the low homogeneous expression of alpha 4 and beta 1 on naive cells. 3) In marked contrast, CD45R0+ cells, generally understood to be memory cells, show not only an overall increase in expression of these integrins (relative to CD45R0- cells) but also heterogeneity. Dramatic heterogeneity is revealed when the markers VLA-alpha 4 and beta 1 are analyzed together. Many CD45R0+ cells show increased levels of both VLA-alpha 4 and VLA-beta 1; however, some have increased levels principally of either VLA-beta 1 or VLA-alpha 4. We hypothesize that T cells becoming memory cells in different microenvironments specialize their integrin phenotype, thereby acquiring distinctive functional and homing capacities; in this process, VLA-4 (CD49d) appears to play a unique role.


Characterization of Dobrava virus: a Hantavirus from Slovenia, Yugoslavia. Small mammals were collected in natural foci of hemorrhagic fever with renal syndrome (HFRS) in Slovenia, Yugoslavia, and a hantavirus was isolated from the lungs of an Apodemus flavicol lis captured in Dobrava village. This new isolate, Dobrava virus, was compared with representative strains of the Hantavirus genus by serological and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. It was found by cross immunofluorescent and enzyme-linked immunosorbent assays that antigenic properties of Dobrava virus were different from those of other hantaviruses. The RNA of this virus was successfully amplified with hantavirus genus reactive primer sets by reverse transcriptase polymerase chain reaction (RT-PCR); however, PCR-RFLP analysis of the amplified product was shown to be unique among those of the known hantaviruses, further indicating that Dobrava virus represents a new hantavirus serotype.


The paired domain-containing nuclear factor pax[b] is expressed in specific commissural interneurons in zebrafish embryos. The zebrafish paired box (Pax) genes are expressed in the early neural tube and are thought to be transcription factors that regulate the differentiation of cells in the central nervous system (CNS). The protein product of one of these Pax genes, pax[b], is detectable as a nuclear antigen in all the regions of the embryo that transcribe the gene including the posterior midbrain, the nephritic primordium, the Wolffian duct, the optic stalk, and, in specific neurons, in the hindbrain and spinal cord. The timing and pattern of axonal outgrowth by the early pax[b]-positive neurons suggest that they are the commissural secondary ascending (CoSA) interneurons in the spinal cord; the primary commissural interneurons (MiD2c and MiD3c) in hindbrain rhombomeres mi2 and mi3; and a previously unclassified set of commissural interneurons that we termed the commissural caudalrhombomere ascending (CoCaA) interneurons in the caudal hindbrain. In contrast, the Mauthner interneurons do not express pax[b] early in development. Shortly after the appearance of the first pax[b]-positive interneurons, additional nuclei adjacent to the first pax[b]-positive interneurons become pax[b] positive. This pattern of expression suggests that the pax[b] protein may be involved in determining the identity of specific commissural interneurons.


Delay in fixation does not affect the immunoreactivity of proliferating cell nuclear antigen (PCNA). The effect of delayed fixation on the immunoreactivity of proliferating cell nuclear antigen (PCNA) was investigated using eight breast carcinomas. Topologically shuffled samples of each tumour were immersed in fixative at times of 0.5, 1, 2, 4, 6, 18, and 24 h after surgical removal. In addition to a PCNA index (percentage of positive cells per 1200 tumour cells), a semi-quantitative PCNA grading system was used, based on estimates of more than or less than 50 per cent of positive tumour cells at each time interval. The PCNA index of six tumours increased by a mean of 10 per cent with a fixation delay of 24 h. The PCNA grade of all eight tumours showed no change with delayed fixation.


Takayasu's arteritis: anatomic change before and after steroid therapy evaluated by angiography and echo-Doppler color-flow. A 36-year-old Italian woman with active Takayasu's disease presented a bilateral occlusions of subclavian artery and stenosis of bilateral common carotid arteries: the maximal diameter stenosis, measured with echo-Doppler color-flow (EDCF) in the longitudinal section was of 43.5 +/- 2.4% on the right and 61 +/- 1.4% on the left. Prednisolone was administered for 30 months at doses from 25 to 6 mg daily (12.5 mg every two days). During steroid therapy we could monitor by EDCF the anatomic change of the involved vessel and a final decrease in carotid wall thickening of 19.8% on the left and 14.0% on the right side. This work demonstrates for the first time that duplex sonography may be an useful tool to asses possible anatomic changes in the carotid lesions of Takayasu's arteritis in response to steroid therapy.


Phenotypic and functional analysis of peripheral blood lymphocytes in oral lichen planus. To assess cellular immunity in oral lichen planus (OLP), peripheral blood mononuclear cells (PBMC) were obtained from 19 OLP patients and 30 control subjects. The proportions of circulating CD45RA+ and CD29+ lymphocyte subsets were determined. The proliferative activity of PBMC to the non-specific plant mitogens phytohemagglutinin (PHA) and concanavalin A (Con A) was examined together with the spontaneous proliferative response and the response in the autologous mixed lymphocyte reaction (AMLR). In the OLP group, the proportion of CD4+ CD45RA+ T lymphocytes was significantly less than control subjects and the proportion of CD4+ CD29+ T lymphocytes was increased significantly. The proliferative response to PHA was similar in OLP and controls subjects. Con A-stimulated PBMC proliferation was decreased significantly in the OLP group. Spontaneous PBMC proliferation in patients with non-reticular lesions was significantly less than control subjects. Despite a mildly depressed response in the AMLR in OLP patients, this result was not statistically significant. Results of the phenotypic analysis of peripheral blood lymphocytes indicate a decreased proportion of naive T cells and an increased proportion of primed memory T cells, although the antigen specificity of these memory cells remains to be determined. Results of the functional assays would seem to reflect this phenotypic shift, and as T cells responding to Con A stimulation and in the AMLR possess suppressor-inducer activity, these results may also suggest an association between OLP and defective innate T cell-mediated suppressor circuits.


Rare expression of the c-erbB-2 oncoprotein in salivary gland tumors: an immunohistochemical study. An immunohistochemical study of c-erbB-2 oncoprotein expression was carried out on 201 cases of primary salivary gland tumors, using a polyclonal antibody, raised to the intracytoplasmic domain of the c-erbB-2 oncogene product. An intense membrane reactivity was observed in one case of sialocarcinoma transformed from pleomorphic adenoma (n = 8) and one case of mucoepidermoid carcinoma (n = 22). A comparative histopathologic evaluation of c-erbB-2 positive tumors showed marked variation in cell size, nuclear pleomorphism, multinucleation, a high mitotic rate and increased lymphoid cell infiltration and an aggressive clinical course with poor survival. The results indicate that c-erbB-2 oncoprotein is rarely expressed in malignant salivary gland tumors. However, the overexpression appears to have a distinct histopathologic feature, but a larger study incorporating histopathology and clinical data would be necessary to correlate the significance of c-erbB-2 oncogene product in salivary malignant tumors.


Scanning electron microscopy of eggs of Mansonia bonneae (Diptera: Culicidae). The egg of Mansonia bonneae (Edwards) is described by means of scanning electron microscopy (SEM) and compared with other sympatric species reported recently in previous studies. Ma. bonneae differed from Ma. uniformis (Theobald), Ma. indiana (Edwards), Ma. annulifera (Theobald), and Ma. annulata (Leicester) in that it has six variably sized dog-tooth spines surrounding the raised central micropyle, and only a single nipplelike stud at the anterior one-third of the egg. A key for identification of the eggs of the five sympatric Mansonia species under SEM is presented.


Identification of human blood in mosquitoes (Diptera: Culicidae) using nonradioactive DNA dot blot hybridization. A dot blot hybridization procedure was developed to detect human blood meals in engorged mosquitoes. A biotinylated DNA probe allowed the detection of 10-100 ng of human DNA, discriminated well between human and nonhuman sources of blood, and cross-reacted only with monkey DNA. Results showed that this method was a specific and sensitive technique for the identification of blood meals up to 100 h after ingestion. The nonisotopic label offers easy handling without the problems inherent in the use of radioisotopes, and it can be adapted for use in routine field tests.


Liposome-mediated modulation of multidrug resistance in human HL-60 leukemia cells. Multidrug resistance (MDR) is a major obstacle in cancer treatment. Resistance of cultured tumor cells to major classes of cytotoxic drugs is frequently due to expression of a plasma membrane P-glycoprotein encoded by MDR genes. We have demonstrated that liposome-encapsulated doxorubicin is more toxic than the free drug and that it modulates MDR in Chinese hamster LZ cells and human colon cancer cells. To investigate further the association between expression of P-glycoprotein and modulation of MDR by liposome-encapsulated doxorubicin, we studied vincristine-resistant HL-60/VCR leukemia cells, which express P-glycoprotein, and doxorubicin-resistant HL-60/ADR leukemia cells, which do not. Cells were exposed to various concentrations of free doxorubicin and liposome-encapsulated doxorubicin. The cellular content of doxorubicin was determined by fluorescence analysis, and cytotoxicity was determined by cell growth inhibition. Photoaffinity-labeling studies of P-glycoprotein binding were performed on HL-60/VCR and HL-60/ADR cells and KB-GSV2 cells transfected with the MDR1 gene (also known as PGY1). The concentrations that caused 50% inhibition of growth (IC50) for free doxorubicin in HL-60, HL-60/ADR, and HL-60/VCR cells were 30 nM, 9 microM, and 0.9 microM, respectively. The values for liposome-encapsulated doxorubicin in parental HL-60 cells and HL-60/ADR cells were 20 nM and 9 microM, respectively, indicating little or no sensitization. In contrast, HL-60/VCR cells were fivefold more sensitive to liposome-encapsulated doxorubicin than to free doxorubicin, and IC50 was reduced to 0.17 microM. In HL-60 cells exposed to liposome-encapsulated doxorubicin, intracellular doxorubicin accumulation was less than that seen with free drug. In contrast, in HL-60/VCR cells, accumulation was twofold to threefold higher than that with free doxorubicin. Liposome-encapsulated doxorubicin completely inhibited the photoaffinity labeling of P-glycoprotein by azidopine in membrane vesicles of HL-60/VCR cells, with a potency comparable to that of azidopine, suggesting that circumvention of MDR by liposomes is related to their specific interaction with P-glycoprotein. The studies with KB-GSV2 cells indicated that blank liposomes can directly inhibit photoaffinity labeling of P-glycoprotein. These results demonstrate the effectiveness of liposome-encapsulated doxorubicin in overcoming resistance in the multidrug-resistant phenotype of HL-60/VCR cells by direct interaction with P-glycoprotein. Furthermore, they indicate that liposome-encapsulated doxorubicin may be an effective treatment for human cancers.


Cerebral ammonia levels and enzyme changes during Plasmodium yoelii infection in mice. Ammonia, lactate and glutamate levels and the activities of glutamine synthetase (GS), glutamate dehydrogenase (GDH), glutaminase (GLN), aspartate transaminase (AST), phosphofructokinase (PFK) and monoamine oxidase (MAO) were compared in the brain tissue of normal and P. yoelii infected mice. The brain lactate increased by 96% at peak parasitaemia. Cerebral ammonia also exhibited an increase in infected mice which was parasitaemia dependent, while glutamate remained almost unchanged. The brain glutamine synthetase registered an increase of 35% (P < 0.001) in post-mitochondrial fractions, this effect being perceptible even at low parasitaemia, but attained constancy at parasitaemia levels higher than 20%. The activity of monoamine oxidase and phosphofructokinase increased by 105% (P < 0.02) and 41% (P < 0.05) respectively while glutamate dehydrogenase decreased by 15% (P < 0.001). Glutaminase and aspartate transaminase were not significantly influenced by infection (tested only at high parasitaemia levels). It has been postulated that cerebral hypoxia and aberrations in ammonia metabolism may both contribute towards malaria induced cerebral complications.


[Clinical significance of posttetanic count (PTC) during onset and spontaneous offset of neuromuscular blockade induced by vecuronium]. To compare the clinical difference of posttetanic count (PTC) during onset and spontaneous offset, the changes of PTC during an intense neuromuscular blockade induced by vecuronium (0.08 mg.kg-1, i.v.) were measured using a neuromuscular transmission analyzer in 64 adult patients anesthetized with nitrous oxide and halothane. Furthermore, intubation score was evaluated when zero PTC was obtained. The obvious movement of diaphragm associated with endotracheal intubation was observed in 9 out of 20 patients (45%) even when it was performed after obtaining zero PTC. The difference of PTC responses between the onset and the offset was found. The difference was that, during the onset, a tetanic response was observed obviously and the height of the posttetanic single twitch was low, and during offset, even when no response was observed, the height of a posttetanic single twitch response was high with the same PTC. These facts indicate that the same PTC has different clinical significance during onset and offset.


[Changes in mental workload and fatigue during performance of a mental task. 1. An experiment in 8-h self-paced transcribing task]. The changes in mental workload and fatigue during a one-day transcribing task were examined by determining some subjective and physiological measures which reflect mental activity. With an interval of one week between the three test days, 12 male students rested and performed self-paced transcribing task with moderate and maximum effort for 8 h each. The subjects transcribed more characters in the task with maximum effort than with moderate effort. In both the morning and afternoon, adrenaline excretion increased and heart rate decreased with the lapse of time. In linear proportion to the total working hours, occipital midline beta-2 amplitude, subjective rating of tiredness and subjective symptoms of fatigue rose, but critical flicker values fell slightly in the task with maximum effort. It was inferred from these results that with lapse of working hours intellectual activity and feeling of fatigue increase and the level of arousal slightly declines. Hence, it was considered that mental workload becomes heavier and subjective feeling of fatigue increases as the working hours become longer.


Protective effect of minaprine against the abnormal changes of 2-deoxyglucose uptake by rat hippocampal slices induced by hypoxia/hypoglycemia. Effect of minaprine on hypoxia- or hypoxia/hypoglycemia (ischemia)-induced impairment of 2-deoxyglucose (2DG) uptake by rat hippocampal slices was evaluated. Since minaprine was found to possess both a stimulating effect on acetylcholine release and a blocking effect on 5-HT2 receptors, the improving effect of minaprine on impaired 2DG uptake was compared to the findings obtained with oxotremorine, ketanserin and pentobarbital. Hippocampal slices were exposed to 20-min ischemia, and then these slices were returned to oxygenated and glucose-containing buffer for 6 hr. Ischemia reduced 30 mM KCl-induced 2DG uptake by the hippocampus. Pretreatment with minaprine, oxotremorine, pentobarbital and ketanserin attenuated the ischemia-induced decline of 2DG uptake. In addition, minaprine, oxotremorine and pentobarbital relatively recovered the increase of 2DG uptake in the hippocampal slices under hypoxia for 45 min. The present results suggest that minaprine exerts a neuroprotective action against ischemia-induced deficit of energy metabolism in vitro.


[Serum adenosine deaminase in human immunodeficiency virus infection. Its relationship with CD4+ lymphocytes and beta 2-microglobulin]. The activity of the deaminase adenosine enzyme (ADA) has principally been related with the functionalism and replication of the T lymphocytes. Its serum behavior and possible clinical use in infection by the human immunodeficiency virus type 1 (HIV-1) was studied. A multicenter study in which the serum values of ADA were examined and compared with those of two reference markers (CD4+ lymphocytes and beta 2-microglobulin) in 35 presumably healthy donors used as controls, in 60 intravenous drug users (IVDU) seronegative for HIV-1, in 69 HIV-1 asymptomatic seropositive intravenous drug users (HIV-1+) and in 48 patients with AIDS. The serum values of ADA were as follows: control group 10.9 +/- 4.2 U/I; IVDU group 17.6 +/- 7.4 U/I; asymptomatic HIV-1+ group 32.7 +/- 10.2 U/I, AIDS group 46.2 +/- 18.2 U/I. Differences between the different groups were statistically significant in themselves and in relation to the control group. A negative correlation was observed (r = 0.47, p < 0.01) with the number of CD4+ lymphocytes and a positive correlation was found with respect to beta 2-microglobulin (r = 0.76, p < 0.001). The values of serum ADA activity in patients with AIDS and tuberculosis (47.4 +/- 17.2 U/I) were not significantly higher (p < 0.05) to those of patients with AIDS without this second infection (45.9 +/- 19.3 U/I). Serum deaminase adenosine may be a useful evolutive marker for human immunodeficiency virus type 1 given that its activity increases significantly in infected patients in agreement with the grade of immunodeficiency and its values correlate well with those of reference markers (CD4+ lymphocytes and beta 2-microglobulin).


Application of magnetic resonance imaging to the measurement of neurodegeneration in rat brain: MRI data correlate strongly with histology and enzymatic analysis. Focal brain ischemia was induced by middle cerebral artery occlusion in the rat. The volume of cerebral damage was determined 2 days later by MRI in vivo and in the same animals histologically. The edema volume as measured by MRI and the histologically determined infarction was highly correlated. As a consequence, the neuroprotective effect of the N-methyl-D-aspartate (NMDA) receptor antagonists CGP 40116 and MK 801 were similar with both methods. Excitotoxic neurodegeneration in the rat striatum was induced by direct injection of quinolinic acid. The degree of damage was evaluated in vivo 1 day later by quantitative MRI, and 7 days later by measuring the activities of neuronal marker enzymes choline acetyltransferase and glutamic acid decarboxylase. Striatal damage assessed using the three approaches was highly correlated. Cerebroprotective efficacy of the NMDA receptor antagonist CGP 40116 was indistinguishable based on all methods. MRI was more reproducible than the enzymatic methods and was faster and simpler than histologic examination for routine analysis of excitotoxic damage and cerebroprotection in vivo in a pharmaceutical research environment.


In vivo identification and monitoring of changes in rat brain glucose by two-dimensional shift-correlated 1H NMR spectroscopy. Intracerebral glucose resonance was directly detected and resolved in vivo by two-dimensional shift-correlated (COSY) 1H NMR spectroscopy in anesthetized rats (n = 4). The relative changes in brain glucose concentration were measured by volume integration of the alpha-D-glucose cross peak in the 2D COSY spectra. This report demonstrates the possibility of monitoring the variations in cerebral glucose following iv injection of glucose.


Glutamate production in islets of Langerhans: properties of phosphate-activated glutaminase. Homogenates of rat pancreas, pancreatic islets, and HIT-T15 cells (a clonal line derived from B cells) catalyzed the breakdown of glutamine to glutamate. This activity was markedly stimulated by the addition of orthophosphate and was much greater in homogenates from islets and the B-cell-derived clonal cell line than in those from whole pancreas. Islet glutaminase was half-maximally stimulated with 40 mmol/L phosphate. Kinetic analyses of the rates of glutamine hydrolysis showed that the Vmax for the reaction increased with the increase in phosphate concentration, whereas the Km for glutamine (2.6 +/- 0.2 mmol/L) was unaltered. The pH optimum for enzyme activity was 8.0 to 8.5 at all phosphate concentrations studied. Glutamine breakdown was enhanced by adenosine triphosphate ([ATP] approximately 100% at 10 mmol/L) and citrate (approximately 30% at 10 mmol/L), but it was unaffected by malate, 2-oxoglutarate, lactate, and ammonia. Glutamate significantly inhibited glutamine hydrolysis. Freshly isolated islets had a low content of both glutamate and glutamine. After culturing for 1 hour in an amino acid-containing medium, the concentrations of glutamine and glutamate increased. Subsequent perifusion without amino acids caused a loss of glutamine and a concomitant increase in glutamate level. Perifusion with 1 mmol/L glutamine led to an increase in both internal glutamine and glutamate. The addition to the perifusion medium of either 10 mmol/L glutamine, 10 mmol/L orthophosphate, or both substantially enhanced insulin release evoked by 10 mmol/L leucine.(ABSTRACT TRUNCATED AT 250 WORDS)


[Mucosa-associated immune system in HIV-1 infection. T-cell subpopulations compared in different segments of the gastrointestinal tract]. T-lymphocyte subsets in duodenal, ileal, coecal, colonal, sigmoidal, and rectal lamina propria were determined in order to compare the distribution of CD2+, CD4+, and CD8+ cell subsets as well as the CD4/CD8 ratio in six different compartments of the gastrointestinal tract. In both HIV1-infected and not HIV1-infected patients, no differences in the distribution of T-lymphocyte subsets could be noted between the compartments examined. The HIV1-infected patients showed a decrease of CD4+ helper cells and CD4/CD8 ratio and an increase of CD8+ suppressor/cytotoxic cells in all examined segments. In terms of lamina propria T-lymphocyte subsets, the intestinal immune system seems to react to HIV1-infection homogenously.


Ammonia regulation of phosphate-activated glutaminase displays regional variation and impairment in the brain of aged rats. The regulation of PAG by ammonia in whole brain (Sprague-Dawley) and regional (Fischer-344) synaptosomal preparations from adult and aged animals was assessed. Whole brain synaptosomal preparations from both age groups displayed a significant decrease in PAG activity with increasing ammonium chloride concentrations, however, the aged rats exhibited a significant attenuation in ammonia-induced PAG inhibition. PAG activity measured in synaptosomes prepared from the striatum (STR), temporal cortex (TCX) and hippocampus (HIPP) was also inhibited by ammonium chloride. The STR showed the greatest degree of ammonia-induced PAG inhibition (55%) followed by the HIPP (30-35%) and the TCX (25-30%). This reduction in PAG activity was significantly attenuated in STR from aged rats at ammonium chloride concentrations greater than 50 microM and in the TCX, PAG activity was significantly attenuated in the aged rats at ammonia concentrations of 0.5 and 1.0 mM. Ammonia regulation of PAG activity in the HIPP appeared to be unaffected by age. Ammonium chloride concentrations up to 5 mM had no effect on GLU release from cortical slices, although GLN efflux was significantly enhanced. These findings suggest that isozymes of PAG may exist in different brain regions based on their differential sensitivity to ammonia. The attenuation of ammonia-induced PAG inhibition seen in aged rats may have deleterious effects in the aged brain.


Guanine nucleotide- and muscarinic agonist-dependent phosphoinositide metabolism in synaptoneurosomes from cerebral cortex of immature rats. Guanine nucleotide-, neurotransmitter-, and fluoride-stimulated accumulation of [3H]inositol phosphates ([3H]InsPs) was measured in [3H]inositol-labeled synaptoneurosomes from cerebral cortex of immature (7-day-old) and adult rats, in order to clarify the role of GTP-binding proteins (G-proteins) in modulating phosphoinositide (PtdIns) metabolism during brain development. GTP(S) [Guanosine 5'-O-(3-thio)triphosphate] time- and concentration-dependently stimulated PtdIns hydrolysis. Its effect was potentiated by full (carbachol, metacholine) and partial (oxotremorine) cholinergic agonists through activation of muscarinic receptors. The presence of deoxycholate was required to demonstrate agonist potentiation of the guanine nucleotide effect. The response to GTP(S) was higher in adult than in immature rats, while the effect of cholinergic agonists was similar at the two ages examined. At both ages, histamine potentiated the effect of GTP(S), while norepinephrine was ineffective. At both ages, guanosine 5'-O-(2-thio)diphosphate [GDP(S)] and pertussis toxin significantly decreased GTP(S)-induced [3H]InsPs formation. The phorbol ester phorbol 12-myristate 13-acetate (PMA), on the other hand, did not inhibit the guanine nucleotide response in synaptoneurosomes from immature rats. NaF mimicked the action of GTP(S) in stimulating PtdIns hydrolysis. Its effect was not affected by carbachol and was highly synergistic with that of AlCl3, according to the concept that fluoroaluminate (AlF4-) is the active stimulatory species. No quantitative differences were found in the response to these salts between immature and adult animals. These results provide evidence that, in both the immature and adult rat brain, neuroreceptor activation is coupled to PtdIns hydrolysis through modulatory G-proteins.


Effect of magnesium on calcium influx activated by glutamate and its agonists in cultured cerebellar granule cells. The effects of Mg2+ on the glutamate-, kainate-, N-methyl-D-aspartate- and quisqualate-induced influx of 45Ca2+ were studied in cultured cerebellar granule cells. The N-methyl-D-aspartate- and quisqualate-evoked influx was totally and the kainate- and glutamate-evoked influx partially blocked in 1.3 mM extracellular Mg2+. The increase in influx induced by kainate, quisqualate and glutamate was maximal at 0.1 mM Mg2+, whereas N-methyl-D-aspartate was most effective in totally Mg(2+)-free media. D-2-Amino-5-phosphonovalerate blocked partially and phencyclidine completely the enhancement of Ca2+ influx by 1 mM quisqualate in 0.1-mM Mg2+ medium. The effect of 10 microM quisqualate was also significantly inhibited by antagonists specific for different glutamate receptor subtypes, including N-methyl-D-aspartate, (RS) alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate and metabotropic receptors. This evidences a heterogeneous action of quisqualate, mediated by different glutamate receptor subtypes in 0.1 mM Mg2+ medium. The efficacy of quisqualate in inducing influx of Ca2+ and the selectivity of antagonists for different receptors are also modified by extracellular Mg2+.


Transglutaminase activity in primary and subcultured rat astroglial cells. Transglutaminases, calcium-dependent thiol enzymes, may be involved in cellular growth control and differentiation, having an intracellular regulatory role in some post-translational modifications found in various classes of proteins. In order to elucidate the involvement of this class of enzymes in cellular differentiation processes, we have assayed transglutaminase activity in primary and subcultured rat glial cells. Reduced activity was found from 3rd to 5th passage. In the 5th passage the activity was some 50% of that found in the primary cultures and was not restored by addition of 10 microM retinoic acid. The decrease of TGase activity, observed during serial passages, could represent an early metabolic alteration related to cell dedifferentiation and loss of growth control. In fact, the subcultured cells may have undergone a "disarranged" state, as confirmed by a decrease in GFAP-stained cells and glutamine synthetase activity, respectively, immunocytochemical and biochemical markers of astroglial cells.


NMDA receptor modification in the fetal guinea pig brain during hypoxia. The effect of maternal hypoxia on the modification of the fetal brain cell membrane N-methyl-D-aspartate (NMDA) receptor and its modulatory sites was investigated. Experiments were conducted in pregnant guinea pigs of 60 days of gestation. Guinea pig fetuses were exposed to maternal hypoxia (FiO2 = 7%) for 60 minutes. Tissue hypoxia in the fetal brain was documented biochemically by decreased levels of ATP and phosphocreatine (91.3% and 88.6% lower than normoxia, respectively). MK-801 binding characteristics (Bmax = number of receptors, Kd = affinity of receptor) were used as an index of NMDA receptor modification. P2 membrane fraction was prepared from the cortex of normoxic and hypoxic fetal brain and washed thoroughly before carrying out the binding assay. In hypoxic brains, Bmax decreased from the normoxic control level 0.79 +/- 0.03 pmol/mg protein to 0.58 +/- 0.03 pmol/mg protein (P < 0.005) and Kd value decreased (increased affinity) from 8.54 +/- 0.27 nM to 4.01 +/- 0.23 nM (P < 0.005) respectively. The MK-801 binding in the absence of added glutamate and glycine in hypoxic brain was 100% higher as compared to controls, indicating an increased sensitivity of the NMDA receptor to activation. The spermine dependent maximum activation of the NMDA receptor increased to 44% in the hypoxic animals as compared to 25% in controls. The Mg2+ response of the NMDA receptor was not affected by hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS)


Modification of modulatory sites of NMDA receptor in the fetal guinea pig brain during development. Ontogeny of the NMDA receptor and modification of its modulatory sites in the developing fetus brain was determined. MK-801 binding characteristics in the presence of glycine, glutamate, Mg2+ and spermine were determined and used as an index of NMDA receptor modification. Experiments were performed on guinea pig fetuses at 30, 45, 50, 55, and 60 days (term = 63 days) of gestation. The Bmax value increased approximately three-fold from 30 days to 60 days of gestation. The Kd value decreased during the 45-50 day period and then increased toward the end of gestation. The Bmax value reached its maximum level by 55 days of gestation, indicating the presence of a maximum number of NMDA receptors by this age, while the apparent affinity of the receptor showed its peak at 45-50 days of gestation, indicating a potential role for NMDA receptor during the proliferation period of brain development in the guinea pig fetus. The activation of NMDA receptor in the presence of glutamate (10 microM) and glycine (10 microM), as measured by MK-801 binding, was absent at 30 days gestation, with the earliest observation occurring at 35 days gestation. The spermine dependent activation decreased with gestational age. Mg2+ ions increased MK-801 binding in the range of 1-20 microM concentration. Sensitivity to Mg2+ dependent activation increased with the gestational age (from 10 microM Mg2+ at 45 days to 2.5 microM at 55 and 60 days).(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of alpha 2- and beta-adrenoceptor agonists on growth hormone secretion following lesion of the noradrenergic system of the rat. The aim of the present investigation was to lesion the noradrenergic system and to measure the effect on growth hormone (GH) secretion following peripheral administration of alpha 2- and beta-adrenoceptor agonists. Direct injection of these agonists into the paraventricular nucleus of the hypothalamus (PVN) and its effect on GH secretion were also investigated. Systemic administration of N-2-chloroethyl-N-ethyl-2-bromobenzylamine (DSP4, 60 mg/kg, injected i.p. 10 days prior to experimentation) significantly decreased the noradrenaline (NA) content of the hippocampus, frontal cortex and hypothalamus but had no effect on the dopamine (DA) or serotonin (5-HT) content of these areas. Bilateral injection of 6-hydroxydopamine (6-OHDA, 10 micrograms/microliters, 14 days prior to experimentation) into the medial forebrain bundle (MFB) caused a greater reduction of NA and also decreased the DA and 5-HT content of the hypothalamus. Analysis of the PVN of the hypothalami of rats following 6-OHDA lesion of the MFB showed significantly decreased NA and 5-HT content. Neither DSP4 treatment nor 6-OHDA lesion of the MFB affected the clonidine (250 micrograms/kg, i.p.) induced stimulation of GH secretion. Injection of isoproterenol (1 mg/kg, i.p.) had varying effects on GH secretion. It stimulated GH release in control rats but not in DSP4 or MFB lesioned rats. Direct injection of clonidine (0.1 microgram/microliters) into the PVN significantly stimulated GH secretion, whereas injection of isoproterenol (2.5 micrograms/microliters) into the PVN did not affect GH levels when compared to controls.(ABSTRACT TRUNCATED AT 250 WORDS)


Glutamate-like immunoreactivity in chick cerebellum and optic tectum. Glutamate was coupled via glutaraldehyde to bovine serum albumin. The conjugate was used for raising specific anti-glutamate antibodies. The purified antibody was used for immunostaining of chick cerebellum and optic tectum. Staining was intense in the molecular layer and in cell bodies of the granule cell layer. In the optic tectum a diffuse staining was detected in the superficial layers of stratum griseum fibrosum superficiale and in cell bodies especially in the layers a and e. Large cell bodies located in the stratum griseum centrale were also stained.


A syndrome of autosomal dominant alternating hemiplegia: clinical presentation mimicking intractable epilepsy; chromosomal studies; and physiologic investigations. We report the familial occurrence and apparent autosomal dominant inheritance of alternating hemiplegia of childhood. The proband, a 9-year-old boy, presented with developmental retardation, rare tonic-clonic seizures, and frequent episodes of flaccid alternating hemiplegia that had been presumed to represent postictal paralysis. The hemiplegia spells, which started in his first year, did not respond to multiple antiepileptics. Between attacks, there was choreoathetosis and dystonic posturing. Father, brother, paternal uncle, and paternal grandmother had similar histories of alternating hemiplegia. Investigations included negative CT, metabolic, and coagulation studies. EEG and SPECT 99mTc exametazime scanning failed to reveal any significant slowing or any major changes in cortical perfusion during hemiplegia as compared with nonhemiplegic periods. The karyotype revealed a balanced reciprocal translocation, 46,XY,t(3;9)(p26;q34) in the patient, in all the affected living relatives, and in one apparently unaffected sibling. The asymptomatic mother had a normal karyotype. Analysis of DNA markers was consistent with the karyotype results. Both affected siblings were treated with and responded to flunarizine therapy, with a greater than 70% decrease in attack frequency. Documented flunarizine trough serum concentrations were 28.9 ng/ml in the proband and 6.6 ng/ml in his brother.


[Comparative study of 24-hour continuous gastric pH-measurements in healthy people and in patients with pancreatic neoplasms treated with duodeno-pancreatic head resection]. The Authors compare pH measurements taken over a 24-h period to monitor, using an original method, a sample group of 11 healthy subject and a series of patients suffering from gastroenteropancreatic diseases. The data obtained confirm the utility of the functional study of gastrointestinal secretion.


[Somatostatin in the treatment of lymphorrhea after lateral neck dissection]. A new pharmacological effect of somatostatin has been verified in the treatment of lymphorrea due to a thoracic duct injury, produced during left lateral neck dissection. The drug (stilamin 3 mcg/kg/h in continuous venous infusion) allowed in 3 case a strongly decrease of the lymphatic loss within 24 hours and the complete depletion within 6 days. Further studies are required to clarify the dynamic effects of the drug in this complication.


[Therapy of arterial hypertension in diabetic patients. When to start the treatment and which drugs to use?]. In this review, the Authors analyzes the more recent data in the Literature about the major cardiovascular risks in diabetic patients affected by systemic arterial hypertension. The more effective pharmacological and not pharmacological treatment is suggested. Particularly the advantages and more important side-effects of antihypertensive drugs are emphasized.


[Empty sella syndrome (ESS) associated with primary hyperparathyroidism. A clinical case and review of the literature]. The report describes a patient with coexisting primary hyperparathyroidism due to adenoma of the left superior lobe and the primary empty sella syndrome (ESS). Pathogenetic mechanisms, clinical pictures, associated illnesses and similarity between these diseases are discussed. A complete diagnostic procedure and follow-up is necessary even if the illnesses are oligosymptomatic. In fact this association may be part of multiple endocrine neoplasia (MEN) or other endocrine diseases.


Simultaneous measurement by HPLC of the excitatory amino acid transmitter candidates homocysteate and homocysteine sulphinate supports a predominant astrocytic localisation. Primary cultures of mouse cerebral cortex neurons, cerebellar granule cells and cortical astrocytes were maintained in vitro for respectively 8-10, 7-10 and 21-24 days. Following these times, amino acids were extracted from the cells by use of ice-cold 70% (v/v) ethanol and the extracts lyophilised. The lyophilised extracts when resuspended were subjected to reverse-phase high performance liquid chromatographic (HPLC) analysis for detection of free amino acids. Samples of cell culture growth medium and water blanks were treated in a similar manner. Identification of L-homocysteate (HCA) and L-homocysteine sulphinate (HCSA) was undertaken by matching retention times with regard to external standards and by 'spiking' cell extracts with authentic compounds. On this basis, HCA and HCSA were consistently detectable in astrocytes at levels of, respectively, 72.3 +/- 33.7 pmol/mg protein (n = 24) and 49.4 +/- 28.7 pmol/mg protein (n = 24). However, in neurons, a peak corresponding to HCSA could not be detected above the background noise, while the area of the peak corresponding to HCA was always greater than, but not significantly different from, that of the background noise present in water blanks. HCA and HCSA were not detectable in the serum used for preparation of the cell culture growth medium. Taken together, these findings indicate a predominant localisation of HCA and HCSA in astrocytes which, at least in culture, appear to possess the metabolic machinery necessary for synthesising and storing these amino acids without any neuronal influence.


Sulmazole effects on PGE2 and D-Ala2-Met-enkephalinamide modulation of cyclic AMP synthesis and neurotransmitter release in a sympathetic ganglion. In the guinea-pig superior cervical ganglion, the Gi blocking agent sulmazole enhanced the basal and prostaglandin E2-induced stimulation of cyclic AMP synthesis but had no effect on the prostaglandin-dependent inhibition of acetylcholine release. On the contrary sulmazole counteracted the inhibitory effect of D-Ala2-Met-enkephalinamide both on cyclic AMP formation and acetylcholine release. Moreover sulmazole eliminated the supra-additive effect of the combination of prostaglandin + opiate on cyclic AMP synthesis. The presence of a Gi-coupled opiate receptor at the pre-and postsynaptic levels is suggested.


Effects of oxiracetam on neurotransmitter release from rat hippocampus slices and synaptosomes. The effects of the nootropic drug oxiracetam on the K(+)-evoked overflow of [3H]D-aspartic acid ([3H]D-ASP), [3H]acetylcholine ([3H]ACh), [3H] gamma-aminobutyric acid ([3H]GABA), [3H]noradrenaline ([3H]NA) and [3H]5-hydroxytryptamine ([3H]5-HT) have been studied in superfused rat hippocampal slices. The overflow of [3H]D-ASP was enhanced by low concentrations of oxiracetam (0.01-1 microM) but not by high concentrations (10-100 microM) which showed some tendency to inhibit it. Similarly, low concentrations of oxiracetam increased, although less effectively, the depolarization-evoked overflow of [3H]ACh, whereas higher concentrations were without effect. At the concentrations active on [3H]D-ASP and [3H]ACh overflow oxiracetam did not affect that of [3H]GABA, [3H]NA or [3H]5-HT. The oxiracetam effects present in slices could not be observed in hippocampal synaptosomes. Thus oxiracetam may selectively increase the release of glutamate and acetylcholine in hippocampus by a mechanism which appears not to be sited in the releasing nerve terminals.


Homocysteate and homocysteine sulfinate, excitatory transmitter candidates present in rat astroglial cultures. The presence of homocysteate and homocysteine sulfinate was demonstrated in extracts prepared from cultures of rat cortical and cerebellar astrocytes as well as from C6 glioblastoma cells by o-phthalaldehyde derivatization and subsequent HPLC analysis. Homocysteate-like immunoreactivity was found in cultured cortical astrocytes by postembedding immunocytochemistry at the level of light microscopy. These findings support the notion of a glial localization of the excitatory transmitter candidate homocysteate.


Co-localization of Met-enkephalin and somatostatin in the spinal cord of the rat. A double-labelling immunofluorescence study of rat spinal dorsal horn was carried out with antisera to Met-enkephalin and somatostatin. Varicosities in laminae I and II were frequently immunoreactive with both antisera, and in addition some neuronal cell bodies in lamina II possessed both types of immunoreactivity. These findings suggest that enkephalin and somatostatin coexist in some axons within the rat superficial dorsal horn and that at least some of these axons are derived from local neurones.


Cholecystokinin- and dopamine-containing mesencephalic neurons provide distinct projections to monkey prefrontal cortex. Retrograde transport and immunohistochemical techniques were utilized to determine if cholecystokinin (CCK)-containing neurons of the primate ventral mesencephalon project to prefrontal cortex, and to examine what relation the CCK innervation of prefrontal cortex bears to the dopaminergic projection to this region. Following injections of Fast blue into monkey prefrontal cortex, retrogradely labeled, CCK-positive neurons were observed predominantly in rostromedial portions of the ventral mesencephalon; these CCK-containing projection neurons were not immunoreactive for tyrosine hydroxylase. Furthermore, dual-labeling studies in the prefrontal cortex revealed that CCK and tyrosine hydroxylase were present in separate populations of axons. These results demonstrate that the CCK innervation of monkey prefrontal cortex arises from both intrinsic and extrinsic sources; in contrast to the rat, the extrinsic CCK innervation of monkey prefrontal cortex is distinct from the dopaminergic mesocortical projection.


Immune control of murine coccidiosis: CD4+ and CD8+ T lymphocytes contribute differentially in resistance to primary and secondary infections. The effect of treatment with monoclonal antibodies (Mabs) which deplete CD4+ or CD8+ T lymphocytes, on infections with Eimeria spp. was examined in NIH mice. Treatment with anti-CD4 Mab increased susceptibility to primary infections with E. vermiformis or E. pragensis and reduced the subsequent resistance of the mice to homologous challenge. Similar treatment of immune mice did not affect their resistance to re-infection but this was reduced in mice depleted of CD8+ T lymphocytes. In mice immunized with E. vermiformis the effect of CD8(+)-depletion was very slight, apparent only as the presence of small numbers of oocysts in the faeces of some mice; in mice immunized with E. pragensis there was a small, though significant, increase in oocyst production, compared with controls and anti-CD4-treated groups. These results confirm the importance of mechanisms involving the function of CD4+ T lymphocytes in the control of primary infections with Eimeria spp. and indicate that CD8+ cells play some part in the expression of resistance to reinfection. They also show that a major part of this resistance was not affected by either of the treatments given.


Dynamic interaction between CD4+ T cells and parasitic helminths: mathematical models of heterogeneity in outcome. Potential mechanisms of immunoregulation have been investigated for the capacity to generate heterogeneity in the outcome of infection with helminth parasites. We have developed a mathematical model of the interaction between T cell and parasite populations, based on the assumption that activation of a Th1 CD4+ T cell response is required for host resistance. Antigen dose-dependent inhibition of Th1 cell proliferation generates heterogeneity in the outcome of host response to infection, with relatively low levels of exposure inducing resistance, and high levels of exposure associated with host susceptibility. Heterogeneity is additionally predicted in the duration of infection before individuals of the resistant class clear infection, with infection becoming more prolonged as the level of exposure rises. Similar categories of response are predicted if an alternative regulatory mechanism, that of interferon gamma-regulated control of Th1 cell differentiation, is substituted into the model. However, the relationship between level of exposure and duration of infection is reversed. Results are discussed in the context of how these simple models of parasite-immune system interactions might be used to make predictions concerning specific examples of parasitic infection.


Age-related standards for T lymphocyte subsets based on uninfected children born to human immunodeficiency virus 1-infected women. The European Collaborative Study. The T lymphocyte subsets in the peripheral blood of 459 uninfected children born to white human immunodeficiency virus 1-infected women included in the European Collaborative Study were measured at regular intervals from birth. More than 2400 observations were used to create smooth age-related reference ranges for CD4 and CD8 counts and percentages, CD4:CD8 ratio and absolute lymphocyte count. Standards are presented for children up to 4 years of age. CD4, CD8 and absolute lymphocyte count rose after birth, peaked at around 6 to 9 months of age and then declined toward adult values. CD4 percentage and CD4:CD8 ratio declined steadily from birth onwards. Centile lines for CD4 count and CD4:CD8 ratio converged markedly with age. For the CD4 values, only 3 to 5% of the variation was attributable to differences between the 10 participating centers. These standards allow T lymphocyte abnormalities to be used more effectively as markers for disease progression and assist in the clinical follow up of human immunodeficiency virus 1-infected children. They also provide a basis for initiating antiretroviral treatment or antimicrobial prophylaxis.


Activation of alpha 1-adrenoceptors modulates the inwardly rectifying potassium currents of mammalian atrial myocytes. The selective alpha 1-adrenergic agonist methoxamine (10(-4)-10(-3) M), in the presence of propranolol (10(-6) M), can reduce both the inwardly rectifying K+ background current (IK1) and the muscarinic cholinergic receptor-activated K+ current (IK,ACh) in rabbit atrial myocytes resulting in action potential prolongation during the final phase of repolarization and a depolarization of the resting membrane potential. The reduction of these K+ currents(s) by alpha 1-adrenoceptor stimulation was insensitive to pre-treatment of atrial myocytes with pertussis toxin (0.15-0.5 micrograms/ml) and was irreversible following intracellular dialysis with the non-hydrolysable guanosine triphosphate (GTP) analogue, Gpp(NH)p (1-5 x 10(-3) M). Neither the protein kinase C (PKC) inhibitors, 1((5-isoquinolinesulphonyl)-2-methylpiperoxine (H-7) (5 x 10(-5) M) and staurosporine (1 x 10(-7) M), nor "downregulation" of PKC by prolonged phorbol ester exposure (5 x 10(-7) M, for 7-8 h) had an effect on the alpha 1-adrenergic modulation of this K+ current. Under cell-attached patch-clamp conditions, bath application of methoxamine reversibly decreased acetylcholine-induced single-channel activity, thus confirming the observed reduction of the ACh-induced current under whole-cell voltage clamp. These results demonstrate that the alpha 1-adrenoceptor, once activated, can reduce current through two different inwardly rectifying K+ channels in rabbit atrial myocytes. These current changes are mediated via a pertussis toxin-insensitive GTP-binding protein, and do not appear to involve the activation of PKC.