Phosphatidylserine-specific phospholipase A1 stimulates histamine release from rat peritoneal mast cells through production of 2-acyl-1-lysophosphatidylserine.

Lysophosphatidylserine (1-acyl-2-lyso-PS) has been shown to stimulate histamine release from rat peritoneal mast cells (RPMC) triggered by FcepsilonRI (high affinity receptor for IgE) cross-linking, although the precise mechanism of lyso-PS production has been obscure. In the present study we show that phosphatidylserine-specific phospholipase A(1), PS-PLA(1), stimulates histamine release from RPMC through production of 2-acyl-1-lyso-PS in the presence of FcepsilonRI cross-linker. The potency of 2-acyl-1-lyso-PS was almost equal to that of 1-acyl-2-lyso-PS. A catalytically inactive PS-PLA(1), in which an active serine residue (Ser(166)) was replaced with an alanine residue did not show such activity. sPLA(2)-IIA, another secretory PLA(2) that is capable of producing lyso-PS in vitro, was also a poor histamine inducer against RPMC. PS-PLA(1) significantly stimulated histamine release from crude RPMC, indicating that lyso-PS is mainly derived from cells other than mast cells. In agreement with this phenomenon, the enzyme stimulated the histamine release more efficiently when RPMC were mixed with apoptotic Jurkat cells. Under these conditions, lyso-PS with unsaturated fatty acid was released from the apoptotic cells treated with PS-PLA(1). Finally, heparin, which has affinity for PS-PLA(1), completely blocked the stimulatory effect of the enzyme. In conclusion, PS-PLA(1) may bind to heparan sulfate proteoglycan, efficiently hydrolyze PS appearing on plasma membranes of apoptotic cells, and stimulate mast cell activation mediated by 2-acyl-1-lyso-PS.

Lysophosphatidic acid (LPA) receptors of the EDG family are differentially activated by LPA species. Structure-activity relationship of cloned LPA receptors.

We examined the structure-activity relationship of cloned lysophosphatidic acid (LPA) receptors (endothelial cell differentiation gene (EDG) 2, EDG4, and EDG7) by measuring [Ca(2+)](i) in Sf9 insect cells expressing each receptor using LPA with various acyl chains bound at either the sn-1 or the sn-2 position of the glycerol backbone. For EDG7 the highest reactivity was observed with LPA with Delta9-unsaturated fatty acid (oleic (18:1), linoleic (18:2), and linolenic (18:3)) at sn-2 followed by 2-palmitoleoyl (16:1) and 2-arachidonoyl (20:4) LPA. In contrast, EDG2 and EDG4 showed broad ligand specificities, although EDG2 and EDG4 discriminated between 14:0 (myristoyl) and 16:0 (palmitoyl), and 12:0 (lauroyl) and 14:0 LPAs, respectively. EDG7 recognizes the cis double bond at the Delta9 position of octadecanoyl residues, since 2-elaidoyl (18:1, trans) and 2-petroselinoyl (18:1, cis-Delta12) LPA were poor ligands for EDG7. In conclusion, the present study demonstrates that each LPA receptor can be activated differentially by the LPA species.

[A case of spontaneous cervical carotid artery dissection managed by using primary stent placement].

The authors report the case of a 35-year-old male who underwent stenting for spontaneous cervical carotid dissection. He presented with sudden onset of hemicrania and left facial palsy followed by left hemiparesis and dysarthria. On admission, carotid angiography revealed postsinus tapering occlusion of the right internal carotid artery. Initially, he was managed with conservative treatment, but his neurological status deteriorated. Findings of brain CT, MRI and IMP-SPECT suggested hypoperfusion of the right cerebrum. A Palmaz stent, 39 mm in length, was successfully placed over the site of the dissection to restore normal patency through the dissected carotid artery. Following stent implantation, his neurological signs improved gradually but completely. Since the procedure, with oral administration of antiplatelet medication, he has suffered no cerebral ischemic events. Follow-up carotid angiography one year after stent implantation showed good patency of the stented segment. The present case emphasizes the usefulness of stenting for spontaneous cervical carotid dissection.

[A case of subarachnoid hemorrhage verified as cerebral vasospasm by using three-dimensional CT angiography (3 D-CTA): reference to comparison with conventional angiography].

The authors present a case of aneurysmal subarachnoid hemorrhage that were verified as cerebral vasospasm by using both three-dimensional CT angioraphy (3 D-CTA) and conventional angiography. A 45-year-old man was referred to our department 4th day after sudden onset of a severe headache. On admission, emergency 3 D-CTA showed the cerebral vasospasm involving M 1 segment. Conventional angiography performed at the same day of the left internal carotid artery confirmed the cerebral vasospasm of the same vessel as 3 D-CTA, and furthermore demonstrated the left middle cerebral artery (MCA) and anterior cerebral artery (ACA) genu aneurysms. The former was seen as a ruptured aneurysm from brain CT findings (Fisher group 3). On the 10th day after the onset, 3 D-CTA demonstrated the remaining severe cerebral vasospasm of the supraclinoid portion of left ICA and M 1 segment. Findings at the conventional angiography subsequently performed were concordant with those of 3 D-CTA. The patient was successfully treated with delayed surgical clipping for both aneurysms without the symptoms related to the cerebral vasospasm and discharged without neurological abnormality. We consider that 3 D-CTA shows promise as a minimally invasive method of evaluating the cerebral vasospasm and would take the place of the conventional angiography.

Molecular cloning and characterization of a novel human G-protein-coupled receptor, EDG7, for lysophosphatidic acid.

Lysophosphatidic acid (LPA), together with sphingosine 1-phosphate, is a bioactive lipid mediator that acts on G-protein-coupled receptors to evoke multiple cellular responses, including Ca(2+) mobilization, modulation of adenylyl cyclase, and mitogen-activated protein (MAP) kinase activation. In this study, we isolated a human cDNA encoding a novel G-protein-coupled receptor, designated EDG7, and characterized it as a cellular receptor for LPA. The amino acid sequence of the EDG7 protein is 53.7 and 48.8% identical to those of the human functional LPA receptors EDG2 and EDG4, respectively, previously identified. LPA (oleoyl) but not other lysophospholipids induced an increase in the [Ca(2+)](i) of EDG7-overexpressing Sf9 cells. Other LPA receptors, EDG4 but not EDG2, transduced the Ca(2+) response by LPA when expressed in Sf9 cells. LPAs with an unsaturated fatty acid but not with a saturated fatty acid induced an increase in the [Ca(2+)](i) of EDG7-expressing Sf9 cells, whereas LPAs with both saturated and unsaturated fatty acids elicited a Ca(2+) response in Sf9 cells expressing EDG4. In EDG7- or EDG4-expressing Sf9 cells, LPA stimulated forskolin-induced increase in intracellular cAMP levels, which was not observed in EDG2-expressing cells. In PC12 cells, EDG4 but not EDG2 or EDG7 mediated the activation of MAP kinase by LPA. Neither the EDG7- nor EDG4-transduced Ca(2+) response or cAMP accumulation was inhibited by pertussis toxin. In conclusion, the present study demonstrates that EDG7, a new member of the EDG family of G-protein-coupled receptors, is a specific LPA receptor that shows distinct properties from known cloned LPA receptors in ligand specificities, Ca(2+) response, modulation of adenylyl cyclase, and MAP kinase activation.

Effects of mepartricin, a polyene macrolide agent, on fecal excretion and serum concentration of estrogen and number of prostatic estrogen receptors in immature rats.

BACKGROUND: Mepartricin, an antifungal agent, was investigated for effects on fecal excretion and serum concentration of sex steroids and the number of sex steroid prostatic receptors in immature rats. METHODS: Mepartricin was orally administered at 2.5, 5, and 10 mg/kg once daily for 2 weeks. Fecal estrogen and testosterone excretions, serum estrogen, testosterone and luteinizing hormone concentrations, and numbers of prostatic estrogen and androgen receptors were assayed. Prostate weight was also monitored. RESULTS: Fecal estrogen excretion showed a dose-dependent increase, which was significant for the two higher dosages. Conversely, the serum estrogen concentration and prostatic estrogen receptors were significantly decreased. No significant changes in fecal testosterone excretion, serum testosterone and luteinizing hormone concentrations, and prostatic androgen receptors were observed. Prostate weight was significantly reduced at 5 mg/kg, but we did not observe dose-dependency. CONCLUSIONS: Mepartricin increases fecal excretion of estrogen by binding with it in the intestinal tract, which results in reducing the serum estrogen concentration and number of prostatic estrogen receptors.

Malignant hypertension associated with obstructive hydrocephalus--case report.

A 36-year-old male presented with headache, vomiting, and gait disturbance. Examination found marked anemia, renal failure, markedly choked disks, and hypertensive encephalopathy. Magnetic resonance imaging demonstrated diffuse swelling of the brainstem and cerebellum, and obstructive hydrocephalus. Treatment with steroid, glycerol, and antihypertensive drugs resulted in a slow decrease in the brain swelling and cerebral edema. However, hydrocephalus and intracranial hypertension persisted, requiring a shunt operation. Hypertensive encephalopathy is usually improved by the treatment of hypertension, but shunt operation may be required to treat exacerbated intracranial pressure associated with obstructive hydrocephalus.

Comparative in-vitro and in-vivo activity of AM-1155 against anaerobic bacteria.

The in-vitro activity of AM-1155, a 6-fluoro-8-methoxy quinolone, was compared with those of temafloxacin, sparfloxacin, tosufloxacin, ciprofloxacin, ofloxacin and cefmetazole, a cephamycin, against a variety of anaerobic bacteria. Although AM-1155 demonstrated only modest activity against the Bacteroides fragilis group and Prevotella bivia (MIC90s > or =3.13 mg/mL), 76% of the B. fragilis strains tested were inhibited at AM-1155 concentrations of 0.78 mg/L. AM-1155 was highly active against Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium spp., Clostridium perfringens and Mobiluncus spp. (MIC90s < or =0.39 mg/L). An in-vivo study using a mixed infection with AM-1155- and tosufloxacin-susceptible B. fragilis and Escherichia coli strains in rat granuloma pouch was performed. AM-1155 was effective against both organisms whereas tosufloxacin was effective only against E. coli. These results correlated well to the higher pouch levels of AM-1155 than those of tosufloxacin. Clostridium difficile overgrowth was found in the caecum of mice treated with ampicillin both 1 and 7 days after 5 days dosing, but not in AM-1155-treated mice. These results suggest that the clinical efficacy of AM-1155 against infections involving most anaerobic bacteria except for the B. fragilis group and P. bivia should be evaluated further.

Posttraumatic ventriculomegaly: hydrocephalus or atrophy? A new approach for diagnosis using CSF dynamics.

Cerebrospinal fluid (CSF) dynamics were correlated to the changes in ventricular size during the first 3 months posttrauma in patients with severe head injury (Glasgow Coma Scale score < or = 8, 75 patients) to distinguish between atrophy and hydrocephalus as the two possible causes of posttraumatic ventriculomegaly. Using the bolus injection technique, the baseline intracranial pressure (ICP), pressure volume index, and resistance for CSF absorption (R0) provided a three-dimensional profile of CSF dynamics that was correlated with ventricular size and Glasgow Outcome Scale (GOS) score at 3, 6, and 12 months posttrauma. Patients were separated into five different groups based on changes in ventricular size, presence of atrophy, and CSF dynamics. Group 1 (normal group, 41.3%) demonstrated normal ventricular size and normal ICP. Group 2 (benign intracranial hypertension group, 14.7%) showed normal ventricular size and elevated ICP. Group 3 (atrophy group, 24%) displayed ventriculomegaly, normal ICP, and normal R0. Group 4 (normal-pressure hydrocephalus group, 9.3%) had ventriculomegaly, normal ICP, and high R0. Group 5 (high-pressure hydrocephalus group, 10.7%) showed ventriculomegaly and elevated ICP with or without high R0. The GOS score in the nonhydrocephalic groups (Groups 1, 2, and 3) was better than in the hydrocephalic groups (Groups 4 and 5). It is concluded from these results that 44% of head injury survivors may develop posttraumatic ventriculomegaly. Posttraumatic hydrocephalus, as identified by abnormal CSF dynamics, was diagnosed in 20% of survivors and their outcome was significantly worse. This study demonstrates the importance of using CSF dynamics as an aid in diagnosis of posttraumatic hydrocephalus and identifying those patients who may benefit from shunt placement.

Evaluation of hydrocephalic periventricular radiolucency by dynamic computed tomography and xenon-computed tomography.

OBJECTIVE: A common finding of computed tomography in a case of normal-pressure hydrocephalus (NPH) is periventricular radiolucency (PVL). We analyzed PVL for patients with hydrocephalus, using dynamic computed tomographic and xenon-computed tomographic techniques to differentiate NPH from similar diseases. METHODS: Dynamic computed tomography was evaluated as a method of diagnosing NPH in 14 patients with computed tomographic findings of both PVL and ventricular dilatation. Of the 14 patients, varying degrees of clinical improvement after shunt surgery were observed in 10 (shunt-effective group) but not in the remaining 4 (shunt-ineffective group). The difference in arrival time between PVL and thalamus, the difference in peak time between PVL and anterior cerebral artery, and cerebral blood flow in PVL by xenon-computed tomographic study were analyzed. RESULTS: The difference in arrival time between PVL and thalamus was significantly longer in the effective group than among the remaining patients. There was no significant difference in PVL/cerebral blood flow and the difference in peak time between PVL and the anterior cerebral artery between the two groups. CONCLUSION: Dynamic computed tomographic analysis of the difference in arrival time between PVL and thalamus is useful for diagnosing NPH and predicting response to shunting.

A case of central pontine myelinolysis after surgical removal of a pituitary tumor.

CASE REPORT: We have experienced a case in which surgical removal of a pituitary tumor from a male patient was followed by the occurrence of hyponatremia, which in turn was later associated with central pontine myelinolysis (CPM). A 4 X 3 X 3 cm pituitary tumor with hypothalamic extension was removed via a transsphenoidal approach. The post-operative course was uneventful until severe hyponatremia developed. To avoid drastic correction of electrolyte levels, reestablishment of normal serum levels was spread over 1 week. Following this, however, various neurologic symptoms such as pseudobulbar palsy, quadriplegia, extrapyramidal symptoms, and mental symptoms appeared. The case was diagnosed as CPM and extrapontine myelinolysis (EPM) on the basis of the clinical course and symptoms, and high-dosage steroid therapy was commenced. RESULTS: There was consequent gradual improvement in symptoms. One month later, MRI revealed typical demyelination lesions in the central pons and striatum. Abnormal electrolyte conditions easily occur in pituitary tumors associated with hypothalamic extension in an altered hormone environment. It is known that CPM and EPM result from drastic correction of hyponatremia. CONCLUSIONS: The frequent measurement of electrolytes and cautious correction of sodium imbalance are important for the prevention of CPM and EPM in the postoperative management of patients who undergo surgery for a pituitary tumor and whose high-dosage steroid therapy are effective.

[In vitro activities of sulopenem, a new parenteral penem, against anaerobes].

In vitro activities of sulopenem, a novel parenteral penem, was compared with those of imipenem, flomoxef, cefuzonam, cefoperazone and sulbactam/ampicillin against 66 reference strains (19 genera, 61 species) and 392 recent clinical isolates of anaerobic bacteria and fastidious aerobic bacteria. Sulopenem had a very broad spectrum against anaerobic bacteria. In general, this compound was active against anaerobic reference strains with MICs of < or = 0.78 micrograms/ml, while being the least active against Bifidobacterium spp. and less active than imipenem against Lactobacillus spp. Sulopenem was more active against Bacteroides fragilis isolates than imipenem and had the highest activities against Bacteroides thetaiotaomicron, Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium spp. and Peptostreptococcus spp. among the antibiotics tested. Sulopenem was not hydrolyzed by oxyiminocephalosporinase type 1 produced by B. fragilis GAI-0558, GAI-7955 and GAI-10150 and its stability was comparable to imipenem. Its susceptibilities to hydrolysis by a metallo-beta-lactamase from B. fragilis GAI-30144 was less than imipenem. Sulopenem (120 mg/kg, 3 times a day for 4 days) was as effective as imipenem/cilastatin against a mixed intraabdominal mice infection due to E. coli and B. fragilis. Sulopenem (20 mg/kg twice a day for 5 days) did not induce an overgrowth of Clostridium difficile in the caecum of mice.

Effects of 3-[N-(2-chlorobenzyl)amino]-6-(1H-imidazol-1-yl) pyridazine dihydrochloride on various aromatase enzyme systems and experimental breast cancer.

The effects of 3-[N-(2-chlorobenzyl)amino]-6-(1H-imidazol-1-yl)pyridazine dihydrochloride (CAS 124070-28-3, MFT-279) on various aromatase enzyme systems and experimental breast cancer were studied. MFT-279 inhibited the aromatase enzyme in vitro with an IC50 value of 2.39 nmol/l. On the other hand, MFT-279 had no effect on cytochrome P-450 dependent reactions of steroid biosynthesis. In pregnant mares' serum gonadotropin (PMSG)-treated female rats, the elevation of ovarian aromatase activity was significantly suppressed by the oral treatment with MFT-279 at 10 and 20 mg/kg. When MFT-279 (20 mg/kg) was orally given to 9,10-dimethyl-1,2-benzanthracene (DMBA)-treated female rats once a day for 28 days, regression of tumors was observed. These results suggest that MFT-279 may be useful for the endocrine therapy of hormone dependent mammary carcinoma.

[Detection of Bacteroides fragilis enterotoxin from extraintestinal clinical isolates of B. fragilis group organisms].

A total of 300 extraintestinal clinical isolates of Bacteroides fragilis group organisms were assayed for enterotoxin using HT29/C1 cells. Of the 179 strains of B. fragilis, 36 (20.0%) were positive for cell culture assay, a positivity which was neutralized by anti-B. fragilis enterotoxin serum; all 36 strains were confirmed to be enterotoxigenic B. fragilis (ETBF). Among B. fragilis isolated from blood, 18 (31%) of 59 strains were ETBF while 18 (15.0%) of 120 strains isolated from non-blood specimens were ETBF (p < 0.05). None of the 121 strains of the B. fragilis group other than B. fragilis were positive for cell culture assay. These results demonstrated that isolation of ETBF from extraintestinal clinical specimens is not uncommon and that B. fragilis enterotoxin, a metalloprotease, may play a role as a pathogenic factor in blood stream infection and sepsis.

Evaluation of the acetazolamide test. Vasoreactivity and cerebral blood volume.

BACKGROUND AND PURPOSE: We evaluated the potential usefulness of the acetazolamide test by investigating whether acetazolamide vasoreactivity reflected the change in resting cerebral blood volume caused by compensatory vasodilation due to a decline in cerebral perfusion pressure. METHODS: We measured resting and acetazolamide-activated cerebral blood flow with a stable xenon-enhanced CT system and resting cerebral blood volume with the subtraction technique using contrast-enhanced CT in 30 patients with various diseases. These parameters were measured in the anterior, middle, and posterior cerebral arterial territories of both hemispheres separately. We evaluated the statistical relationships between resting cerebral blood volume and vasoreactivity in these three territories, and the significance of the correlations was tested by ANOVA/ANCOVA to adjust for the double entries. RESULTS: Significant negative linear relationships were demonstrated between the resting cerebral blood volume and the change in cerebral blood flow, expressed as a percentage induced by acetazolamide activation, for the anterior (r = -.607, P = .0004), middle (r = -.551, P = .0015), and posterior (r = -.523, P = .0078) cerebral arterial territories and between the resting cerebral blood volume and the increase in cerebral blood flow (absolute values) for the anterior (r = -.512, P = .0164) and middle (r = -.523, P = .0001) but not the posterior (r = -.571, P = .0563) cerebral arterial territories. CONCLUSIONS: The acetazolamide test appears to be useful for the investigation of compensatory vasodilation: the vasoreactivity can be calculated as the increased cerebral blood flow expressed as a percentage or an absolute value, which both reflect cerebral blood volume directly.

Antibiotic susceptibility profiles of Bacteroides fragilis and Bacteroides thetaiotaomicron in Japan from 1990 to 1992.

The antimicrobial susceptibility of clinical isolates of Bacteroides fragilis and Bacteroides thetaiotaomicron collected from December 1990 through November 1992 was determined by the agar dilution technique. Metronidazole, imipenem, and cefoperazone/sulbactam remained highly active against both organisms. Rates of resistance to those agents were 0, 2%, and 0.9% in B. fragilis and 0, 0.9%, and 3% in B. thetaiotaomicron, respectively. Cefoxitin and other cephamycins were active against B. fragilis; rates of resistance to these agents did not tend to increase. With the inclusion of these data, the variation of rates of resistance to several agents was summarized for each year from 1987 to 1992. Rates of resistance to imipenem decreased in 1991 and 1992 among isolates of B. fragilis (2.3% in 1991, 1.8% in 1992) and B. thetaiotaomicron (2.4% in 1991, 0 in 1992). Rates of resistance to cefoxitin in B. thetaiotaomicron varied from 10% to 38% during these 6 years, though the distributional peak of MIC values did not change. The rate of resistance to ofloxacin in B. fragilis increased from 42% in 1989 to 81% in 1992. The rate of resistance to ampicillin in B. thetaiotaomicron was 68% in 1992--approximately 30% lower than before. Mostly, however, the rates of resistance to the antimicrobial agents examined did not change significantly.

Sphingolipid composition in Bacteroides species.

Sphingolipid profiles of strains from species of genus Bacteroides, and representative strains from Prevotella and Porphyromonas, were analyzed by thin-layer chromatography and infrared spectrophotometry. Two major types of phosphosphingolipid, ceramide phosphorylethanolamine and ceramide phosphorylglycerol, were detected in B. fragilis, B. ovatus, B. uniformis, B. caccae, B. eggerthii, B. thetaiotaomicron, and B. stercoris, but not in B. merdae, B. distasonis, and B. vulgatus. Strains from the genera Prevotella and Porphyromonas also contained these two sphingolipids. These sphingolipid profiles were conserved within the species tested, and may be useful for differentiation and recognition of relationships within the genera Bacteroides, Prevotella and Porphyromonas.

Cerebrovascular carbon dioxide reactivity assessed by intracranial pressure dynamics in severely head injured patients.

Appropriate management of intracranial pressure (ICP) in severely head injured patients depends in part on the cerebral vessel reactivity to PCO2; loss of CO2 reactivity has been associated with poor outcome. This study describes a new method for evaluating vascular reactivity in head-injured patients by determining the sensitivity of ICP change to alterations in PCO2. This method was combined with measurements of the pressure volume index (PVI), which allowed calculation of blood volume change necessary to alter ICP. The objective of this study was to investigate the ICP response and the blood volume change corresponding to alterations in PCO2 and to examine the correlation of responsivity and outcome as measured on the Glasgow Outcome Scale. The PVI and ICP at different end-tidal PCO2 levels produced by mild hypo- and hyperventilation were obtained in 49 patients with Glasgow Coma Scale scores of less than 8 and over a wide range of PCO2 (25 to 40 mm Hg) in eight patients. Given the assumption that the PVI remained constant during alteration of PaCO2, the estimated blood volume change per torr change of PCO2 was calculated by the following equation: BVR = PVI x delta log(ICP)/delta PCO2, where BVR = blood volume reactivity. The data in this study showed that PVI remained stable with changes in PCO2, thus validating the assumption used in the blood volume estimates. Moreover, the response of ICP to PCO2 alterations followed an exponential curve that could be described in terms of the responsivity indices to capnic stimuli. It was found that responsivity to hypocapnia was reduced by 50% compared to responsivity to hypercapnia measured within 24 hours of injury (p < 0.01). The sensitivity of ICP to estimated blood volume changes in patients with a PVI of less than 15 ml was extremely high with only 4 ml of blood required to raise ICP by 10 mm Hg. The authors conclude from these data that, following traumatic injury, the resistance vessels are in a state of persistent vasoconstriction, possibly due to vasospasm or compression. Furthermore, BVR correlates with outcome on the Glasgow Coma Scale, indicating that assessment of cerebrovascular response within the first 24 hours of injury may be of prognostic value.