TACR1 gene polymorphism and sex differences in postoperative nausea and vomiting.

We hypothesised that the genetic effect of single nucleotide polymorphisms in the TACR1 gene, which encodes NK1 receptors, could influence the sex difference in postoperative nausea and vomiting. Thirty-two selected single nucleotide polymorphisms were genotyped by the Sanger sequencing method in 200 patients who underwent lower abdominal surgery. The incidence and severity of postoperative nausea and vomiting were evaluated after surgery. The rs3755468-SNP showed significant association with the incidence and severity of postoperative nausea and vomiting (p = 0.016). The TT haplotype defined by two single nucleotide polymorphisms, including the rs3755468-SNP, was associated with reduced incidence and severity of postoperative nausea and vomiting in female patients (p = 0.03). The rs3755468-SNP is located within the predicted oestrogen response element and a DNase I hypersensitive site. The single nucleotide polymorphisms in the TACR1 gene are associated with sex differences in postoperative nausea and vomiting and may help to elucidate the mechanisms underlying these differences.

Improved non-invasive total haemoglobin measurements after in-vivo adjustment.

We hypothesised that an in-vivo adjustment method and/or a newer sensor would increase the accuracy of non-invasive and continuous haemoglobin monitoring (SpHb) measurements. Two sensors, the R1-25 and R2-25a (the newer version), were used with laboratory total haemoglobin concentration (tHb) values simultaneously recorded. In-vivo adjusted SpHb (AdHb) was calculated by a simple formula: AdHb = SpHb - (1(st) SpHb - 1(st) tHb). The correlation coefficients between SpHb (or AdHb) and tHb were compared: SpHb in both sensors correlated strongly with tHb (p < 0.0001). In-vivo adjustment improved the correlation coefficient between SpHb and tHb from 0.86 to 0.95 for the R1-25 and from 0.83 to 0.93 for the R2-25a. There was no difference between the R1-25 and R2-25a sensors. The in vivo adjustment method improved the accuracy of SpHb measurements in both sensors.

C3b receptor (CR1) on erythrocytes in various diseases.

Complement receptor for C3b (CR1) on erythrocytes was investigated in various diseases by immune adherence hemagglutination (IAHA) using aggregated human IgG. In normal controls, 21 out of 312 (6%) revealed defective CR1 reactivity, and there was no difference in the prevalence of defective CR1 reactivity between female (11/157, 7%) and male (10/155, 6%). Among diseases examined significantly high prevalence of defective reactivity of CR1 on erythrocytes was seen in systemic lupus erythematosus (SLE) (22/30, 73%) and malignancy of hematopoietic system, especially in acute myelogenous leukemia (AML)(6/11, 55%).

Molecular-genetic study of Duchenne and Becker muscular dystrophies: deletion analyses of 45 Japanese patients and segregation analyses in their families with RFLPs based on the data from normal Japanese females.

This study consisted of 1) molecular deletion analyses in patients with Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) using the entire cDNA for the DMD gene as hybridization probes, 2) RFLP analyses in a large number of Japanese normal women using 11 DMD-linked cloned DNAs as probes, and 3) segregation analyses with these RFLP data in 17 DMD families in which prenatal or carrier diagnosis was required. The deletion study showed that 18 (43%) of 42 male DMD patients had a deletion within the DMD gene, while no detectable deletion was found in 3 BMD patients. These deletions were preferentially observed at the 5' end of the DMD gene, while no deletion was found in the 3' portion of the gene. Of a total of 15 RFLPs detected with the 11 probes, one was a new RFLP (probe/enzyme: P20/MspI). In 6 RFLPs, the allele frequencies in the Japanese were statistically different from those in the Caucasian. Based on the RFLP data combined with the result of the deletion study, an estimated diagnostic rate for prenatal diagnosis and/or carrier detection in the Japanese DMD families was 63%. The real diagnostic rate obtained from the prenatal and carrier diagnoses, which were practically performed in 17 families, corresponded to the estimation. A protocol useful for the diagnosis in Japanese DMD families is presented.

Vascular endothelial cell injury and platelet embolism in Duchenne muscular dystrophy at the preclinical stage.

Blood vessels in muscle biopsy specimens from 4 Duchenne muscular dystrophy (DMD) patients (including 3 at the preclinical stage) were examined by electron microscopy and compared with those in non-diagnostic biopsy specimens from age-matched controls and cases of other childhood neuromuscular disorders. The most striking feature was the blister-like swelling of vascular endothelial cells in the biopsied muscle specimens from the 3 preclinical stage DMD patients, which was observed in 23-39% of the small blood vessels examined. Other noticeable features in the preclinical DMD patients were: (1) replication of the basement membrane, there being more than 3 layers in 30% of the capillaries; (2) many degenerating and regenerating capillaries; and (3) platelet adhesion and aggregation in small blood vessels including small arteries and veins. Morphometric analysis showed that the capillary and endothelial cell areas were much greater in the preclinical DMD patients than in the controls or the cases of the other neuromuscular disorders. These phenomena strongly suggest an as yet undetermined process in blood vessels in preclinical DMD.

PCR and immunoblot analyses of dystrophin in Becker muscular dystrophy.

The dystrophin gene was examined by PCR analysis in 30 Japanese patients with Becker muscular dystrophy (BMD). Fifteen PCR of these patients had exon deletion, generally, less than three exons. Muscle biopsies were also performed in 20 BMD patients (10 with sequence deletions, and 10 without detectable sequence deletions) in order to correlate PCR findings with immunoblot and immunostaining data. A patchy, heterogeneous membrane immunostaining pattern of reduced intensity was found, irrespective of the presence or absence of deletions. Immunoblotting studies demonstrated dystrophin of low molecular mass and quantity in BMD patients with deletion mutations, while a low quantity of dystrophin with an apparent wild type molecular mass was observed in nearly half the BMD patients without detectable deletions. However, these dystrophins were also found to have slightly abnormal molecular masses when the standard electrophoresis time was prolonged. This suggests that immunoblots and PCR data correlate well in patients with BMD. Additionally, it is suggested that immunoblot assays can detect abnormalities in dystrophin in the absence of detectable PCR deletions.

Vascular alterations in Fukuyama type congenital muscular dystrophy.

Blood vessels in muscle biopsy specimens from 6 Fukuyama type congenital muscular dystrophy (FCMD) patients were examined by electron microscopy and compared with ones in non-diagnostic biopsy specimens from age-matched controls and patients with childhood neuromuscular disorders. The most striking feature was the blister-like swelling of vascular endothelial cells in the biopsied muscle specimens from 5 of the 6 patients with FCMD. Morphometric analysis of capillaries in biopsied muscles showed the extremely greater capillary, endothelial and pericyte areas in the FCMD patients than in controls. These phenomena are quite similar to those found in Duchenne muscular dystrophy (DMD) at the preclinical stage and suggest an as yet undetermined process in blood vessels in FCMD as well as DMD. An immunohistochemical study involving dystrophin antibodies showed positive staining in FCMD.

Creatine kinase brain isoenzyme in infantile osteopetrosis.

A 14-month-old girl with infantile osteopetrosis had hematologic and neurologic complications with severe brain atrophy. Although serum contained high creatine kinase brain isoenzyme activity (CK-BB), CK-BB activity was not detected on repeated cerebrospinal fluid examinations. After frequent blood transfusions and steroid therapy, hematologic involvement improved gradually and disappeared finally at age 11 months; serum CK-BB tended to show a concomitant proportional increase in activity. A 111Indium chloride scan was performed at age 4 weeks when the patient had relatively low serum CK-BB activity. It indicated active extramedullary hematopoiesis in the liver and spleen. The second scan was performed at age 12 months when she had high serum CK-BB activity and indicated active medullary hematopoiesis in the cranium. The tests disclosed that the elevated serum CK-BB activity was the result of bone marrow serum leakage, and not leakage from brain tissue. This finding may be a good marker of medullary hematopoietic activity in patients with osteopetrosis. Meanwhile, biopsied sural nerve revealed storage of cellular debris, including myelin figures in the Schwann cells, which suggested increased degradation process in the cells or lysosomal enzyme deficiency.

Infantile osteopetrosis complicating neuronal ceroid lipofuscinosis.

An autopsy case of infantile osteopetrosis complicating neuronal ceroid-lipofuscinosis is reported. Autopsy revealed generalized sclerosis and thickening of cortical and spongy bones, formation of mineralized cartilagenous tissues, and narrowing of the marrow cavities associated with decreased hematopoietic cell components. Around the thickened bone trabecles, osteoclasts lacked a ruffled border and clear zone along the cell membrane facing the bone matrix surface. The brain was markedly atrophic with neuronal cell loss and focal gliosis, and the remaining neuronal cells accumulated brown granular pigments, which were confirmed histochemically and electron-microscopically to be ceroid and lipofuscin. In the cerebral medulla, the development of myelin sheaths was extremely poor. Also, the occurrence of Lex type glycolipids and GM3 and the apparent absence of cerebroside and cerebroside sulfate were proved by biochemical analysis, suggesting that the brain was still in a stage of embryonic development or still in the process of differentiation. Except for one suggestive case, this is the first case of complicating neuronal ceroid-lipofuscinosis in infantile osteopetrosis.

Selective IgM deficiency in a patient with Hashimoto's disease.

A 70-year-old man with Hashimoto's disease had selective IgM deficiency, while other immunoglobulin levels were normal. In vitro mixing experiments were carried out in which B cells and T cells from the patient and from a healthy control donor were co-cultured in the presence of pokeweed mitogen, in order to investigate the etiology of IgM hypoproduction. The results indicated that the patient had B-cell dysfunction, involving the impairment of B-cell differentiation. In addition, both IgG of the healthy control donor and thyroid hormone may play important roles in the pathogenesis of this case.

Immunomodulators and the complement system.

The possible role of immunomodulators in the host-defense mechanism against neoplasm is discussed from the standpoint of the complement system. Serum complement is activated by the majority of immunomodulators in vitro via either the classical or the alternative pathway, and this activation is sometimes observed following systemic administration of immunomodulators. Besides activating the complement, systemic administration of immunomodulators elevates serum complement levels, and in some cases binds complements to tumor cells. Activated serum complement by an immunomodulator induced an accumulation of PMNs in ascites with tumor cell destruction when intraperitoneally injected, indicating that complement-derived chemotactic factors C3a and C5a generated by an immunomodulator had an antitumor effect. This evidence strongly supports the concept that complement system plays an important role in the host-defense mechanism against neoplasm.

[Strategy for molecular diagnosis of Duchenne muscular dystrophy: alleles frequency in RFLPs detected with X-linked probes in normal Japanese].

Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder. Recently it has been suggested that the protein named "dystrophin" is not produced in the DMD muscle because of their mutant of defective DMD gene. Recent molecular studies using subclones from the DMD-cDNA as probes revealed a DNA deletion in about a half of the DMD patients. Since not all patients showed deletion, segregation analysis with restriction fragment length polymorphisms (RFLPs) is still necessary and important for diagnosis not only patients but female carriers as well. Therefore, we tried to detect RFLPs with the use of many probes localized within and/or flanked with the DMD gene and made a protocol for the diagnosis with our RFLP data. Among 50 normal Japanese females, the frequencies of rare alleles which identified with enzyme/probe combinations in each were as follows (RFLPs practically available for diagnosis are underlined): 0.38 for p782/EcoRI; 0.14 for p99-6/Pst I: 0.45 for pJ66-HI/Pst I: 0.16 for pERT87-30/Bgl II; 0.44 for 87-15/Xmn I: 0.07 for 87-15/Taq I; 0.33 for 87-8/Taq I; 0.44 for 87-1/Xmn I; 0.45 for 87-1/Mva I; 0.43 for Xj-1.1/Taq I; 0.34 for 754-11/EcoR I; 0.41 for 11.28/Taq I. Some of our results were different from the Caucasian both on the allele frequencies and the restriction fragment length. We proposed a diagnostic protocol of DMD and it's carriers in Japan. Firstly, segregation analysis should be performed with five different combinations of double probe/enzyme, such as Yj-1,.1/L1.28/Taq I, ERT87-1/87-15/Xmn I, 87-1/Mva I, J66-HI/99-6/Pst I, and 782/754-11/EcoR I.(ABSTRACT TRUNCATED AT 250 WORDS)

[A case of acute exacerbation of idiopathic interstitial pneumonia after pneumonectomy].

A 52-year-old male was scheduled for right upper lobectomy due to lung cancer. The patient was diagnosed as having stable idiopathic interstitial pneumonia (IIP) based on preoperative lung biopsy, physical examination and laboratory data. He received prophylactic steroids to prevent an acute exacerbation of IIP. However, he fell into severe hypoxia 5 days after operation, and died of respiratory failure 43 days after operation. It must be born in mind that the patients with IIP may suffer exacerbation even if their lungs have been diagnosed as in stable stage. Prophylactic use of steroids may cause a deterioration in IIP due to delayed healing and infection.

[Cesarean section using a temporary IVC filter in a pregnant woman with several histories of deep vein thrombosis].

A 31-year-old pregnant woman with a history of several episodes of deep vein thrombosis was scheduled for Caesarian section. The patient had an increased risk of intraoperative pulmonary thromboembolism due to thrombi isolated from deep veins. This is because a thrombus can be formed easily in deep veins by the influence of hypercoagulability in the third trimester and by the disturbance of venous return due to the pressure of the enlarged pregnant uterus. A temporary IVC filter (Filtre Thery, France) was inserted preoperatively. The filter basket was set in the IVC cephalad from the left renal vein and caudad from the hepatic vein. The operation was completed without any trouble. Rehabilitation was safely initiated just after the surgery by insertion of the filter and then the filter was removed on the third postoperative day. We conclude that the preoperative insertion of a temporary IVC filter is a useful technique to prevent intraoperative pulmonary thromboembolism in a patient with a history of deep vein thrombosis.

[Two siblings with interstitial deletion of chromosome 14 [46 XX, del (14) (q12 q13.3)]].

Two siblings with interstitial deletion of chromosome 14 not associated with ring formation were reported. Clinical features of the patients included failure to thrive, severe mental retardation, microcephaly, round face, hypertelorism, micrognathia and high-arched palate. They were common also in five previously reported cases. Other peculiar finding was hyperthyrotropinemia in their neonatal periods; mild hypothyroidism was found in the elder sister, and hyperthyrotropinemia was only transient in the younger sister. By a high-resolution G-banding analysis, the maternal origin of the chromosome with the deletion was noted.

[The relationship between complement C3 receptors (CR1, CR3) on polymorphonuclear leukocytes and complement fragments during hemodialysis].

The expression of complement receptor type 1 (CR1) and type 3 (CR3) on polymorphonuclear leukocytes (PMNs) and generation of complement fragments, C3a, C5a, C4d, iC3b and Bb, were studied in patients during hemodialysis using cuprammonium rayon (Cu) membranes. Furthermore, the relation between the expression of CR1 and CR3 on PMNs from healthy donors and complement fragments was investigated. The expression of CR1 and CR3 on PMNs increased during hemodialysis. Plasma C3a, C5a and iC3b levels increased in the first 15 minutes and then decreased at 120 minutes of dialysis. But plasma Bb level remained high until the end of hemodialysis. Purified Bb had no effect on the expression of CR1 and CR3 on PMNs, but C5a augmented those expression in vitro. Nafamostat mesilate, an artificial proteinase inhibitor, inhibited augmentation of complement receptors on PMNs in concentration dependent fashion. C5a generated through the activation of complement was thought to take an important role in the increased expression of CR1 and CR3 on PMNs.

[Plasma levels of complement fragments during hemodialysis in patients with chronic renal failure].

The kinetics of hemodialysis-induced leukopenia and generation of complement fragments including C3a, C5a, C4d, iC3b and Bb were investigated in 14 patients during hemodialysis using cellulose acetate (CA), cuprophan (Cu) and ethylenevinyl alcohol (EVA) membranes. A marked leukopenia in the first 15 minutes was observed in CA and Cu. Plasma C3a levels were higher in CA than in Cu and EVA. Plasma C5a levels were higher in CA and Cu than in EVA. There was a negative correlation between the white blood cell counts and plasma C5a levels at 15 minutes (gamma = -0.85, p less than 0.001). Plasma C4d levels showed no increase in all membranes. Plasma iC3b levels were higher significantly in Cu than in CA and EVA. Plasma Bb levels in the first 15 minutes increased significantly in all membranes, and furthermore continued to increase till the end of hemodialysis in CA and Cu. This study revealed that all the membranes tested activated the complement via the alternative pathway to produce Bb, iC3b, C3a and C5a. C5a was thought to take an important role in transient leukopenia. Furthermore, Bb was accumulated during hemodialysis in CA and Cu, and its biological effects on patients undergoing hemodialysis should be studied.

[Kinetics of immune complex deposition and influence of decomplementation on their clearance in cationized antigen induced acute serum sickness].

Using a murine acute serum sickness model caused by cationized bovine gamma-globulin (CBGG), the histological findings, accumulation of CBGG in the organs and the effect of decomplementation for the clearance of immune complex (IC) were investigated. The accumulation of CBGG increased in the lungs in the presence of anti-CBGG antibody 1 hour after injection of CBGG, but did not change in the kidneys. This suggests that CBGG forms IC in situ in the kidneys, and that circulating IC accumulates in the lungs. Decomplementation did not influence the uptake of CBGG immediately after challenge but delayed the clearance of CBGG in the kidneys, lungs, liver and spleen. A beneficial role of the complement system in the clearance of IC even for those formed in situ was recognized. Histological changes, cellular infiltration and microvascular destruction, evoked in the lungs in this experimental model immediately after challenge, were not seen in the kidney, suggesting the different modes of complement activation in these organs.

[Changes of complement in immune complex glomerulonephritis induced by cationized antigen].

Complement system takes an important role in the pathogenesis of immune complex glomerulonephritis. In this study the precise changes of serum complement and its deposition in the glomeruli were investigated in the mice models induced with cationized bovine gamma-globulin (CBGG). Balb/c mice preimmunized with CBGG were injected intravenously with CBGG three times every 24 hours. Proteinuria and hypoalbuminemia have developed from day 3 until the death on day 9-13 of progressed azotemia. This nephritis resembled to mesangial proliferative glomerulonephritis histopathologically. Immunofluorescent study revealed granular deposits of CBGG, IgG, A, M and C3 along the glomerular capillary loop. Serum C3 decreased to about 50% of normal controls within 1 hour after challenge of antigen and plasma C3 conversion evaluated by crossed-immunoelectrophoresis, occurred in accordance with C3 reduction and then decreased. Immunofluorescent study showed a big difference in the mode of deposition among IgG, CBGG and C3. The most intense deposits of C3 in the glomeruli were seen 12 hours after challenge of CBGG, and then gradually decreased until the next challenge. IgG and CBGG antigen deposited most intensely soon after the challenge of antigen, and gradually decreased. These results led us to conclude that there existed two phases of complement activation in this immune complex induced mice glomerulonephritis; an early phase activation of complement by antigen-antibody complexes in the circulation and later phase activation of complement by immune complexes after deposition of them in situ glomeruli.

[Role of oxygen species and lipid peroxidation for anti-tumor effect of intra-arterial injection with degradable starch microspheres].

The role of superoxide and lipid peroxidation for anti-tumor effect of intra-arterial injection with degradable starch microspheres (DSM) was investigated in rabbits. The anti-tumor effect of intra-arterial injection with DSM was studied in rabbits with VX2 carcinoma of the hind leg. The tumor growth in rabbit treated with DSM 5 times was completely suppressed, and thiobarbituric acid (TBA)-reactive substances in the tumor tissue treated with DSM were significantly increased. But the anti-tumor effect of DSM and the increase in TBA reactive substances in the tumor tissue treated with DSM were significantly inhibited by treatment with superoxide dismutase combined with catalase. These results suggest that the anti-tumor effect of intra-arterial injection with DSM may be due to ischemia-reperfusion injury and that active oxygen species and lipid peroxidation may play an important role in the anti-tumor effect of intra-arterial injection with DSM.