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Purpose: Preoperative medical management is critical to prevent intraoperative cardiovascular complications in patients with pheochromocytomas and paragangliomas (PPGLs). Initial treatment involves α-adrenergic receptor blockers. However, while the routine use of metyrosine alongside these blockers is not strongly recommended due to a lack of evidence supporting its efficacy and associated safety concerns, there are previous studies on combination therapy with phenoxybenzamine and metyrosine. There are few reports on combination therapy with the selective α1-adrenergic receptor blocker doxazosin. Therefore, we investigated this combination treatment, which theoretically can affect perioperative outcomes in patients with PPGLs. To our knowledge, this is the first such study. Methods: This retrospective single-center observational study involved 51 patients who underwent surgical resection of PPGLs at Kobe University Hospital between 2014 and 2022. All patients received doxazosin at maximum doses. Fourteen patients received concomitant metyrosine, while 37 received doxazosin alone. Their perioperative outcomes were compared. Results: No severe event, such as acute coronary syndrome, was observed in either group. Intraoperatively, the doxazosin + metyrosine group exhibited a lower median minimum systolic blood pressure (56 [54–60] vs.68 [59–74] mmHg, P= 0.03) and required lower median remifentanil (P= 0.04) and diltiazem (P= 0.02) doses than the doxazosin-alone group. Conclusion: The combination of metyrosine and doxazosin as a preoperative treatment for PPGLs affects intraoperative circulatory hemodynamics, such as a reduced occurrence of blood pressure elevation during surgery. Further research is necessary to identify patients who will benefit most from this combination treatment.

Introduction: Levels of serum selenium (Se) and zinc (Zn) decrease when total parental nutrition (TPN) is administered without trace element supplementation for just a few weeks. These trace elements are involved in thyroid hormone metabolism and their deficiencies cause thyroid dysfunction. However, there have been few reports on the details of its clinical course. Case presentation: A 50-year-old man presented with thyroid dysfunction due to Se and Zn deficiency. He had an approximately 70-cm residual small intestine after undergoing intestinal resection and he received TPN without trace element supplementation for one and a half months. Blood tests revealed high levels of thyroid-stimulating hormone (TSH) and free thyroxine (FT4) and low levels of free triiodothyronine (FT3). An abnormal pattern of thyroid function led to suspicion of Se deficiency. Se supplementation raised FT3 levels and lowered FT4 levels to within their respective reference ranges; however, subclinical hypothyroidism persisted with transient TSH elevation. We suspected that Zn deficiency also contributed to the hypothyroidism and, therefore, initiated Zn supplementation, which resulted in normalization of thyroid function. Discussion: Although thyroid dysfunction has been reported in many studies conducted on Se and Zn deficiencies, hormonal patterns vary between reports. Further accumulation of cases, including detailed data on nutritional status, would be of benefit to elucidate the clinical reality. Conclusion: It is important to consider Se and Zn deficiencies when TSH and FT4 levels are elevated. It should also be noted that transient TSH elevation may be observed with Se supplementation.

Although weight loss programs including diet and exercise targeting patients with obesity are essential, the percentage of patients who successfully achieve weight loss is less than 10%. One cause of failure to achieve weight loss is a strong sense of hunger, which is difficult to sustain. Additionally, it is important to prevent a decrease in muscle mass. Various protein supplements have been used to overcome these problems. However, there is insufficient evidence to support this theory. This study aimed to evaluate the effect of soy or whey protein beverages in conjunction with general dietary and exercise therapies on weight loss.

One of the major foods causing norovirus gastroenteritis is bivalve shellfish, such as oysters. Depuration and relaying methods have been used to control norovirus. However, these methods may be inadequate to control norovirus gastroenteritis. The present study aimed to investigate the effectiveness of high hydrostatic pressure (HHP) treatment in controlling norovirus in shelled oysters, by evaluating the inactivating effect of HHP on murine norovirus strain 1 (MNV-1) inoculated into a buffer, oyster homogenate, and shelled oysters. First, MNV-1 was inoculated (infectivity of 4.5 log PFU/mL) into the buffer and oyster homogenate, with a pH of 6.3 and salinity (NaCl) of 1.5%, mimicking the habitats of the Pacific oyster (Crassostrea gigas). HHP treatment at 100, 200, 275, and 300 MPa for 2 and 5 min was conducted at an initial temperature of 0 or 5°C. The infectivity of MNV-1 in both the buffer and the oyster homogenate was lower when the initial temperature was 0°C. In the buffer, the infectivity of MNV-1 decreased to 1.8 log PFU/mL after HHP treatment (200 MPa for 5 min at 0°C), and the inactivating effect was higher in the buffer than in the oyster homogenate. MNV-1 was inoculated into shelled oysters (4.8 log PFU per oyster), and HHP treatment was done at 275, 300, and 350 MPa for 5 min at the initial temperature of 0°C. The infectivity of MNV-1 decreased to 2.8 log PFU per oyster after HHP treatment at 275 MPa for 5 min. The results indicate that the inactivating effect of HHP treatment varies, depending on the medium surrounding the viral particles. Inactivation was best in buffer, followed by oyster homogenate and shelled oysters. The data could inform the development of methods to control norovirus in oysters.

Sessile serrated adenoma/polyp (SSA/P) is considered as an early precursor in the serrated neoplasia pathway leading to colorectal cancer development. The conventional adenoma-carcinoma sequence is associated with activation of the WNT signaling pathway, although its role in serrated lesions is still controversial. To clarify differences in WNT signaling activation in association with MLH1 methylation or BRAF/KRAS mutations between serrated and conventional routes, we performed β-catenin immunostaining, methylation-specific PCR for MLH1 and WNT signaling associated genes such as AXIN2, APC, and MCC and secreted frizzled-related proteins (SFRPs), and direct sequencing of BRAF/KRAS in 27 SSA/Ps, 14 SSA/Ps with high-grade dysplasia and 9 SSA/Ps with submucosal carcinoma, as well as 19 conventional adenomas, 26 adenomas with high-grade dysplasia and 25 adenomas with submucosal carcinoma. Nuclear β-catenin labelings were significantly lower in the serrated series than in their adenoma counterparts, and a significant increment in those labelings was found from SSA/Ps to those with high-grade dysplasia or submucosal carcinoma. The frequency of MLH1 and SFRP4 methylation was significantly higher in SSA/P series, as compared with corresponding adenoma series. AXIN2 and MCC were more frequently methylated in SSA/Ps with high-grade dysplasia and those with submucosal carcinoma than in adenoma counterparts. Stepwise increment of AXIN2 and MCC methylation was identified from SSA/Ps through those with high-grade dysplasia to those with submucosal carcinoma. A significant correlation was seen between nuclear β-catenin expression and methylation of AXIN2 or MCC in the SSA/P series. BRAF mutation was more frequent, whereas KRAS mutation was less frequent in the SSA/P series as compared with the adenoma series. There was an inverse association of BRAF mutation with AXIN2 methylation in SSA/P series. In conclusion, WNT/β-catenin signal activation mediated by the methylation of SFRP4, MCC, and AXIN2 may make different contributions to colorectal neoplasia between the serrated and conventional routes.

Sessile serrated adenoma/polyp (SSA/P) is considered as an early precursor in the serrated neoplasia pathway leading to colorectal cancer development. The conventional adenoma-carcinoma sequence is associated with activation of the WNT signaling pathway, although its role in serrated lesions is still controversial. To clarify differences in WNT signaling activation in association with MLH1methylation or BRAF/KRASmutations between serrated and conventional routes, we performed β-catenin immunostaining, methylation-specific PCR for MLH1and WNT signaling associated genes such as AXIN2, APC, and MCCand secreted frizzled-related proteins (SFRPs), and direct sequencing of BRAF/KRASin 27 SSA/Ps, 14 SSA/Ps with high-grade dysplasia and 9 SSA/Ps with submucosal carcinoma, as well as 19 conventional adenomas, 26 adenomas with high-grade dysplasia and 25 adenomas with submucosal carcinoma. Nuclear β-catenin labelings were significantly lower in the serrated series than in their adenoma counterparts, and a significant increment in those labelings was found from SSA/Ps to those with high-grade dysplasia or submucosal carcinoma. The frequency of MLH1and SFRP4methylation was significantly higher in SSA/P series, as compared with corresponding adenoma series. AXIN2and MCCwere more frequently methylated in SSA/Ps with high-grade dysplasia and those with submucosal carcinoma than in adenoma counterparts. Stepwise increment of AXIN2and MCCmethylation was identified from SSA/Ps through those with high-grade dysplasia to those with submucosal carcinoma. A significant correlation was seen between nuclear β-catenin expression and methylation of AXIN2or MCCin the SSA/P series. BRAFmutation was more frequent, whereas KRASmutation was less frequent in the SSA/P series as compared with the adenoma series. There was an inverse association of BRAFmutation with AXIN2methylation in SSA/P series. In conclusion, WNT/β-catenin signal activation mediated by the methylation of SFRP4, MCC, and AXIN2may make different contributions to colorectal neoplasia between the serrated and conventional routes.

Iron (Fe) is an essential nutrient in both plants and humans. Fe deficiency on calcareous soil with low Fe availability is a major agricultural problem. Nicotianamine (NA) is one of the Fe chelator in plants, which is involved in metal translocation into seeds, and serves as an antihypertensive substance in humans. In this study, soybean plants overexpressing the barley NA synthase 1 (HvNAS1) gene driven by the constitutive CaMV 35S promoter were produced using Agrobacterium-mediated transformation. The transgenic soybean showed no growth defect and grew normally. The NA content of transgenic soybean seeds was up to four-fold greater than that of non-transgenic (NT) soybean seeds. The level of HvNAS1expression was positively correlated with the amount of NA, and a high concentration of NA was maintained in the seeds in succeeding generations. The Fe concentration was approximately two-fold greater in transgenic soybean seeds than in NT soybean seeds. Furthermore, the transgenic soybeans showed tolerance to low Fe availability in calcareous soil. Our results suggested that increasing the NA content in soybean seeds by the overexpression of HvNAS1offers potential benefits for both human health and agricultural productivity.

Fluorescent combination: Cell‐penetrating probes derived from the diterpene fusicoccin can form ternary complexes with 14–3–3 proteins and phosphopeptide ligands, whereupon the probes site‐specifically attach a fluorescent tag onto the surface of the 14–3–3 proteins.

Fluoreszierende Kombination: Vom Diterpen Fusicoccin abgeleitete zellpenetrierende Sonden bilden zusammen mit Phosphopeptid‐Liganden und 14‐3‐3‐Proteinen ternäre Komplexe, woraufhin die Sonden auf der Oberfläche der 14‐3‐3‐Proteine positionsspezifisch einen Fluoreszenzmarker anbringen.

Abstract: In humans, iron (Fe) and zinc (Zn) deficiencies result in major worldwide health problems. Transgenic technologies to produce Fe- and Zn-biofortified rice varieties offer a promising potential solution. Nicotianamine, the precursor of phytosiderophores, chelates Fe2+ and Zn2+ and plays an important role in transporting these metals to both vegetative and reproductive organs within the plant. Our objective was to increase Fe and Zn contents in rice grains by overexpressing the barley nicotianamine synthase gene HvNAS1. HvNAS1-overexpressing transgenic rice showed increased HvNAS1 expression and subsequent increases in endogenous nicotianamine and phytosiderophore content in shoots, roots, and seeds. Fe and Zn concentrations in polished T1 seeds from transgenic plants increased more than three and twofold, respectively; Fe and Zn concentrations also increased in both polished and brown T2 seeds. These results suggest that the overproduction of nicotianamine enhances the translocation of Fe and Zn into rice grains.

Iron deficiency is a major cause of reduced crop yields worldwide, particularly in calcareous soils. Unlike barley, rice is highly susceptible to iron deficiency because of a low capacity to secrete mugineic acid family phytosiderophores (MAs), which are iron chelators secreted by graminaceous plants. We present an approach toward the generation along with field trials of transgenic rice lines exhibiting increased tolerance to iron deficiency. Cloning barley genes that encode biosynthetic enzymes for MAs enabled us to produce transgenic rice plants by introducing barley MAs biosynthesis-related genes. We tested three transgenic lines possessing barley genomic fragments responsible for MAs biosynthesis in a paddy field experiment on calcareous soil, which revealed tolerance of these lines to low iron availability. We also applied new approaches to generate iron-deficiency-tolerant rice lines, including the introduction of an engineered ferric-chelate reductase gene and manipulation of transcription factor genes regulating the iron deficiency response.

Helicobacter pylori (H. pylori)is one of the most common bacterial pathogens in humans but the route of transmission remains unclear. We investigated transmission by DNA fingerprinting analysis of cultured H. pylorifrom pediatric patients and their family members.

Increasing the iron (Fe) and zinc (Zn) concentrations of staple foods, such as rice, could solve Fe and Zn deficiencies, which are two of the most serious nutritional problems affecting humans. Mugineic acid family phytosiderophores (MAs) play a very important role in the uptake of Fe from the soil and Fe transport within the plant in graminaceous plants. To explore the possibility of MAs increasing the Fe concentration in grains, we cultivated three transgenic rice lines possessing barley genome fragments containing genes for MAs synthesis (i.e., HvNAS1, HvNAS1, and HvNAAT-Aand HvNAAT-Bor IDS3) in a paddy field with Andosol soils. Polished rice seeds with IDS3inserts had up to 1.40 and 1.35 times higher Fe and Zn concentrations, respectively, compared to non-transgenic rice seeds. Enhanced MAs production due to the introduced barley genes is suggested to be effective for increasing Fe and Zn concentrations in rice grains.

A highly sensitive quantitative method for analyzing nicotianamine (NA) by liquid chromatography/electrospray ionization time-of-flight mass spectrometry (LC/ESI-TOF-MS) is reported. Fluorenylmethoxycarbonylation of nicotianamine reduced its polarity and enabled its retention in a reversed-phase column. The adoption of Nε-nicotyllysine (NL) as an internal standard ensured reliable quantification by giving a linear calibration curve drawn between the NA/NL molar ratios of standard solutions injected and the NA/NL area ratios in mass chromatograms. The high sensitivity of this analytical method allowed us to measure the amount of NA. This analytical method has applications to all research concerning NA.

Nicotianamine (NA), a metal chelator, is ubiquitous in higher plants. In humans, NA inhibits angiotensin I-converting enzyme (ACE), and consequently reduces high blood pressure. Nicotianamine is synthesized from the trimerization of S-adenosylmethionine (SAM) by NA synthase (NAS). Here, we aimed to produce large amounts of NA fermentatively by introducing the Arabidopsis AtNAS2gene into Saccharomyces cerevisiaestrain SCY4. This strain can accumulate up to 100 times the usual amount of SAM, and this is considered desirable for overproduction of NA. Nicotianamine was produced in the engineered yeast, and the NA level increased with incubation time until the stationary phase. The maximum concentration of intracellular NA obtained was 766±33 μg/g wet weight. Successful production of NA in S. cerevisiae should pave the way for industrial production of this novel antihypertensive substrate.

ABSTRACTRecent studies have speculated on the possible role of the mother in transmitting Helicobacter pyloriinfection to their children. In an attempt to either prove or disprove this supposition, we investigated the rates of infection of children born to H. pylori-positive mothers from birth to 5 years of age using serology and the stool antigen test. When infection of the children did occur, the strains from the children were compared to those of their mothers using DNA analysis. Sixty-nine of the 350 pregnant mothers (19.7%) had a positive serology for H. pylori. Fifty-one children underwent serological examinations and stool antigen tests at 4 to 6 days after birth, followed by 1, 3, and 6 months. They were continuously given the stool antigen test at 4- to 6-month intervals until the age of 5 years. Gastric juice samples were collected from the infected children and their mothers for culture and DNA analyses using a random amplified polymorphic DNA fingerprinting method. None of the 51 children acquired H. pyloriinfection during the first year of life. Of the 44 children enrolled in a 5-year follow-up study, five (11%) acquired H. pyloriinfection. They acquired the infection at the age of 1 year 2 months, 1 year 3 months, 1 year 6 months, 1 year 8 months, and 4 years 4 months. Random amplified polymorphic DNA fingerprinting confirmed that the strains of the five children exhibited DNA fingerprinting patterns identical to those of their mothers. These findings suggest that mother-to-child transmission is the most probable cause of intrafamilial spread of H. pylori.

Iron (Fe) is an essential microelement for plant growth. Fe availability is particularly limited on calcareous soils, which have high pH. Approximately 30% of the world's soils are considered calcareous with low Fe availability, which results in extensive areas of Fe deficiency in plants. Some graminaceous plants are known to secrete high amounts of mugineic acid family phytosiderophores (MAs) under Fe deficiency. This Fe acquisition system is called the Strategy-II mechanism. Tolerance to Fe deficiency in graminaceous plants is thought to depend on the quantity of MAs secreted by plants under Fe deficiency stress. This system was utilized to enhance the tolerance of rice to low Fe availability. Transgenic rice expressing the barley naatgenes, one of the genes for the enzymes on the biosynthetic pathway of MAs, showed tolerance to low Fe availability when grown in a calcareous soil.

t(11;18) is a recurrent chromosomal abnormality observed in mucosa-associated lymphoid tissue (MALT)-type lymphoma. API2 and MLT genes have been implicated. The authors devised a dual-color interphase fluorescence in situ hybridization (FISH) system to detect splitting of 11q22 and its fusion with 18q21. Subjects were 44 cases of extranodal lymphoma and cases of primary macroglobulinemia. Whenever RNA was available, reverse transcriptase–polymerase chain reaction followed by sequence analysis was performed. Positive cases by dual-color FISH analysis were restricted to MALT-type lymphoma and one case of primary macroglobulinemia. Among 24 cases of MALT-type lymphoma, 14 (58) (4 gastric, 5 pulmonary, 3 orbital, 1 salivary, and 1 thyroid lymphomas) had splitting of the 11q22 region probes and fusion of signals suggesting the translocation of chromosome 11 and 18. Reverse transcriptase–polymerase chain reaction analysis showed the API2/MLT gene fusion in 9 of 10 cases. Sequence analyses showed three different modes of involvement of the MLT gene, whereas the breakpoint at API2 was the same. Monoclonal component of serum immunoglobulin M was observed in 3 of 14 positive cases for the translocation. Direct visualization using dual-color FISH on samples serves as a molecular tool for management of MALT-type lymphoma with API2/MLT gene fusion.

1The mechanisms by which 5‐hydroxytryptamine (5‐HT) depresses transmitter release from lamprey reticulospinal axons were investigated. These axons make glutamatergic synapses onto spinal ventral horn neurons. 5‐HT reduces release at these synapses, yet the mechanisms remain unclear.2Excitatory postsynaptic currents (EPSCs) evoked by stimulation of reticulospinal axons were recorded in ventral horn neurons. 5‐HT depressed the EPSCs in a dose‐dependent manner with an apparent Kmof 2.3 μm.3To examine the presynaptic effect of 5‐HT, electrophysiological and optical recordings were made from presynaptic axons. Action potentials evoked Ca2+transients in the axons loaded with a Ca2+‐sensitive dye. 5‐HT slightly reduced the Ca2+transient.4A third‐power relationship between Ca2+entry and transmitter release was determined. However, presynaptic Ca2+currents were unaffected by 5‐HT.5Further, in the presence of a K+channel blocker, 4‐aminopyridine (4‐AP), 5‐HT left unaltered the presynaptic Ca2+transient, ruling out the possibility of its direct action on presynaptic Ca2+current. 5‐HT activated a 4‐AP‐sensitive current with a reversal potential of ‐95 mV in these axons.6The basal Ca2+concentration did not affect 5‐HT‐mediated inhibition of release. Although 5‐HT caused a subtle reduction in resting axonal [Ca2+]i, synaptic responses recorded during enhanced resting [Ca2+]i, by giving stimulus trains, were equally depressed by 5‐HT.75‐HT reduced the frequency of TTX‐insensitive spontaneous EPSCs at these synapses, but had no effect on their amplitude. We propose a mechanism of inhibition for transmitter release by 5‐HT that is independent of presynaptic Ca2+entry.

Mugineic acid family phytosiderophores (MAs) secreted from 19 Fe deficient wild and ancestral species of cultivated wheats were analyzed by HPLC to clarify whether wheat plants have or have not the ability to produce and secrete MAs other than deoxymugineic acid (DMA). Analysis of root washings showed that all the species with a different genome type secreted DMA but not mugineic acid (MA) or hydroxymugineic acid (HMA). This showed that all the species possessing A, B, S or D genomes have the ability to synthesize DMA. The results that even the ancient species do not secrete MA and HMA showed that wheat species have not had the ability to secrete MA and HMA from the beginning of their evolution.In addition to DMA, an unknown MAs was detected in the root washings of Triticum dicoccum (AABB) and Triticum aestivum cv. N61 (AABBDD). Traces of the compound was also detected in Aegilops bicornis (SbSb) and all the species with AABBDD genome. Except Aegilops bicornis (SbSb), all the species that secreted the unknown MAs have B genome, and SbSb is proposed to be somewhat related to B genome. Thus the unknown MAs might have originated from a B genome.

Recent studies have suggested an association of Helicobacter pyloriand iron‐deficiency anemia (IDA). This is a report of six cases of IDA associated with H. pylorigastritis. Six patients with IDA were studied (5 boys and 1 girl; mean age 13.6 years; range 13–15 years). Five had a medical history of long‐standing IDA and of oral iron supplementation at outpatient clinics. The anemia recurred after the iron therapy had been discontinued. The sixth patient was admitted to the hospital with severe IDA. An extensive work‐up was ordered that included technetium‐99m (99mTc) scans for Meckel's diverticulum, total colonoscopy, and gastrointestinal endoscopy. After biopsy‐based H. pyloritest results were confirmed to be positive, anti‐H. pyloritherapy consisting of lansoprazole, clarithromycin, and metronidazole was administered for 2 weeks with no iron supplementation. The hematologic profile and iron status were assessed periodically after the end of the eradication regimen. Upper gastrointestinal endoscopy revealed a marked antral nodularity but no evidence of bleeding lesions in all the patients. Given the histology and the fact that rapid urease test results were positive, chronic active gastritis with H. pyloriwas diagnosed in all these cases. H. pyloriwas successfully eradicated in all the patients. There was no evidence of IDA in any of the follow‐up examinations between 27 and 50 months after anti‐H. pyloritherapy. H. pyloriinfection may be involved in cases of IDA of unknown origin, and the eradication of H. pylorican be associated with the resolution of anemia.

Bone is one of the essential target tissues of growth hormone (GH). In bone remodeling, cell-matrix attachment is important where focal adhesion kinase (FAK) is involved. FAK plays a central role in determining the shape and motility of cells in response to the extracellular matrix stimuli. In the present study, we have demonstrated that GH stimulated tyrosine phosphorylation of FAK in human osteoblast-like cells, Saos2. Moreover, GH rapidly enhanced the formation of actin stress fibers. In Saos2, Jak2 was tyrosine phosphorylated by GH stimulation, and AG490, a Jak2 specific inhibitor, inhibited GH-induced tyrosine phosphorylation of FAK and actin stress fiber reorganization. These results suggest that GH activates FAK via Jak2, and stimulates the formation of actin stress fibers in Saos2. Activation of FAK and actin stress fiber formation induced by GH seem to be important for the physiological role of osteoblast.

1The role of cystine‐glutamate exchange in controlling the extracellular glutamate concentration in the central nervous system was examined by whole‐cell clamping neurons in rat brain slices, and using their glutamate receptors as sensors of extracellular glutamate concentration.2Applying cystine to cerebellar slices generated a membrane current in Purkinje cells which was abolished by glutamate receptor blockers. Similar cystine‐evoked currents were seen in pyramidal cells of frontal cortex slices.3Control experiments on non‐N‐methyl‐D‐aspartate (non‐NMDA) receptors in enzymatically isolated Purkinje cells showed that cystine did not produce a current in slice Purkinje cells by directly activating glutamate receptors, nor by potentiating the action of background levels of glutamate on receptors. Experiments on isolated salamander Müller cells showed that cystine did not block Na+‐dependent GLAST glutamate transporters (homologous to the transporters in the Bergmann glia ensheathing the Purkinje cells), nor did it block the current produced by EAAT4 and EAAC1 glutamate transporters in Purkinje cells. Thus the cystine‐evoked current in Purkinje cells is not due to a rise in extracellular glutamate concentration caused by block of Na+‐dependent uptake.4The dependence of cystine‐evoked current on cystine concentration in slice Purkinje cells could be fitted by a Michaelis‐Menten relation with a Kmof 250 μM. The Kmpredicted from this for cystine activating glutamate efflux is less than 140 μM, because of the non‐linear dependence on glutamate concentration of the Purkinje cell current. The current evoked by 1 mM cystine was little affected by removal of extracellular chloride or addition of 1 mM furosemide (frusemide), but was potentiated by 1 mM 4,4′‐diisothiocyanatostilbene‐2,2′‐disulfonic acid (DIDS).5These data suggest that external cystine generates a current in slice Purkinje cells by activating cystine‐glutamate exchange in cells of the slice, releasing glutamate which activates non‐NMDA receptors in the Purkinje cell membrane.

In the livers of patients whose sera contained antibodies to C100-3 antigen (anti-HCV) and hepatitis C virus (HCV) RNA, the presence of HCV RNA and HCV capsid protein (CP) antigen was demonstrated byin situ hybridization and immunohistochemistry, respectively. It was found that occasional hepatocytes in four of ten livers from patients whose sera were positive for both anti-HCV and HCV RNA hybridized with antisense as well as sense oligonucleotide DNA probes, whereas the probes did not hybridize with livers from patients whose sera were negative for anti-HCV and HCV RNA. Monoclonal antibody against a synthetic oligopeptide with amino acid sequence of HCV CP reacted with occasional hepatocytes in six of 14 livers from patients whose sera contained these HCV markers, but not with livers from patients whose sera were negative for both of them. These results suggest that HCV proliferates within hepatocytes since both antisense and sense probes hybridized with cytoplasm of the hepatocytes and that the virus matures in the cytoplasm as the capsid proteins were also found in the hepatocytes.

Anti-c100-3 (Ortho) was determined in the sera of 152 patients with HBs antigen-positive chronic liver diseases to assess coinfection of hepatitis B virus (HBV) and hepatitis C virus (HCV). Eleven patients (7.2%) were positive for anti-c100-3. Anti-CP-9 (Okamoto) and HCV-RNA (RT-PCR) were also examined in these 11 patients. Anti-CP-9 was detected in 7 patients and HCV-RNA was detected in all 11 patients. Four of the 11 anti-c100-3-positive patients were positive for HBe antigen (HBeAg) and others were negative. In 8 of the 11 patients, HCV was suspected to be superinfected by blood transfusion. In HBeAgpositive patients, serum glutamic pyruvic transaminase (SGPT) was elevated in relation to active replication of HBV shown by DNA-polymerase activity. The histological findings showed chronic active hepatitis, with or without cirrhosis. On the other hand, in HBeAg-negative patients, SGPT fluctuated without evidence of active replication of HBV. Active inflammation in the liver was observed in 3 of 5 HBeAg-negative patients by liver biopsy. These findings suggest that HBV might play an important role in chronic active inflammation in HBeAg-positive patients coinfected with HCV, and that HCV might be responsible for continuous inflammation in HBeAg-negative patients coinfected with HCV.

To identify intrafamilial transmission of hepatitis C virus, we tested for antibodies to hepatitis C virus (anti-HCV) in serum samples of 160 family members (12 parents, 17 siblings, 73 children, and 58 spouses) of 76 index patients with chronic hepatitis C. Anti-HCV was detected in 14 of 160 family members (8.75%). This positive rate was significantly higher than the anti-HCV-positive rate in age-matched volunteer blood donors in our district (8.75% vs. 1.42%; P<0.01). Among 14 members, 5 had normal liver function tests and 9 had hepatitis C. In 4 of 73 children, anti-HCV was detected (5.48% vs. 0.55%; P<0.01). In the two children, transmission from mother to child in their childhood was suspected, and in the other children, transmission was from father to child. Furthermore, 7 of 58 spouses (12.1%) had anti-HCV (12.1% vs. 2.13%; P<0.01). These results suggest that intrafamilial transmission, especially horizontal infection may play an important role in the spread of hepatitis C.

Polysaccharide fractions of the leaves of a number of Aloe species significantly reduced blood sugar level in mice. Activity-directed fractionation of the polysaccharide fraction from Aloe arborescens Mill. var. natalensis (Wood, et Ev.) Bgr. leaves yielded two glycans, arborans A and B, which elicited marked hypoglycemic effects in normal and alloxan-induced hyperglycemic mice.