A new clustering model based on the seminal plasma/serum ratios of multiple trace element concentrations in male patients with subfertility.

Purpose: To investigate whether seminal plasma (SP)/serum ratios of multiple trace elements (TEs) can classify patients with male subfertility. Methods: SP/serum ratios of 20 TEs (lithium, sodium, magnesium, phosphorus, sulfur, potassium, calcium, manganese, iron, cobalt, copper, zinc, arsenic, selenium, rubidium, strontium, molybdenum, cesium, barium, and thallium) were calculated for healthy volunteers (n = 4) and those consulting for male subfertility (n = 245). Volunteer semen samples were collected by split ejaculation into early and subsequent fractions, and SP/serum ratio data were compared between fractions. The patients' SP/serum ratio data were used in an unsupervised clustering analysis and qualitatively compared with the data from the fractions of ejaculation from the volunteers. Semen quality parameters and pregnancy outcomes were compared between patient clusters. Results: The early fraction of volunteers was characterized by lower phosphorus and arsenic and 18 other higher TEs than the subsequent fraction. Cluster analysis classified patients into four distinct clusters, one sharing characteristics with the early fraction and another with the subsequent fraction. One cluster with the early fraction characteristics had significantly lower semen volume and higher pregnancy rates from spontaneous pregnancies or intrauterine insemination. Conclusions: Classification of patients based on SP/serum ratios of multiple TEs represents the dominance of fractions of ejaculation samples.

Successful Desensitization with Crizotinib after Crizotinib-induced Liver Injury in ROS1-rearranged Lung Adenocarcinoma.

Crizotinib has been approved for patients with advanced lung adenocarcinoma harboring rearrangements of the c-ROS-1 (ROS1) and anaplastic lymphoma kinase (ALK) genes. We report a patient with ROS1-rearranged lung adenocarcinoma who developed a crizotinib-induced mixed/cholestatic type of liver injury. The patient discontinued crizotinib after 34 days due to liver toxicity. Twenty-four days later, when transaminases and C reactive protein (CRP) were normalized, crizotinib was resumed using an oral desensitization method. The patient was successfully treated for manageable recurrence of liver injury and has been able to continue the treatment.

C-Glycosyltransferases catalyzing the formation of di-C-glucosyl flavonoids in citrus plants.

Citrus plants accumulate many kinds of flavonoids, including di-C-glucosyl flavonoids, which have attracted considerable attention due to their health benefits. However, the biosynthesis of di-C-glucosyl flavonoids has not been elucidated at the molecular level. Here, we identified the C-glycosyltransferases (CGTs) FcCGT (UGT708G1) and CuCGT (UGT708G2) as the primary enzymes involved in the biosynthesis of di-C-glucosyl flavonoids in the citrus plants kumquat (Fortunella crassifolia) and satsuma mandarin (Citrus unshiu), respectively. The amino acid sequences of these CGTs were 98% identical, indicating that CGT genes are highly conserved in the citrus family. The recombinant enzymes FcCGT and CuCGT utilized 2-hydroxyflavanones, dihydrochalcone, and their mono-C-glucosides as sugar acceptors and produced corresponding di-C-glucosides. The Km and kcat values of FcCGT toward phloretin were <0.5 µm and 12.0 sec-1 , and those toward nothofagin (3'-C-glucosylphloretin) were 14.4 µm and 5.3 sec-1 , respectively; these values are comparable with those of other glycosyltransferases reported to date. Transcripts of both CGT genes were found to concentrate in various plant organs, and particularly in leaves. Our results suggest that di-C-glucosyl flavonoid biosynthesis proceeds via a single enzyme using either 2-hydroxyflavanones or phloretin as a substrate in citrus plants. In addition, Escherichia coli cells expressing CGT genes were found to be capable of producing di-C-glucosyl flavonoids, which is promising for commercial production of these valuable compounds.

Krtap11-1, a hair keratin-associated protein, as a possible crucial element for the physical properties of hair shafts.

BACKGROUND: The physical properties of the hair are predominantly determined by the assembly of keratin bundles. The keratin-associated proteins (Krtaps) are thought to be involved in keratin bundle assembly, however, the functional role of the individual member still remains largely unknown. OBJECTIVE: The aim of this study is to clarify the role of a unique class of Krtaps, Krtap11-1, in the development and physical properties of the hair. METHODS: The expression regulation of Krtap11-1 was analyzed and its binding partners in the hair cortex were determined. Also, the effects of the forcible expression of this protein on the hair follicle development were analyzed in culture. RESULTS: The expression pattern of Krtap11-1 was concentrically asymmetric in the faulty hair that develops in Foxn1nu mice. In cultured keratinocytes, the expression of Krtap11-1 transgene product was strictly regulated by the keratinization process and proteasome-dependent protein elimination. While the association with keratin as well as the cohesive self-assembly of Krtap11-1 appeared to be stabilized by disulfide cross-links, the biotinylated Krtap11-1 probe enabled the adherence to certain type I keratins in the hair cortex, including K31, 33 and 34, in the absence of disulfide formation. When embryonic upper lip rudiments were forcibly introduced with Krtap11-1, the hair follicles formed irregularly arranged globular hair keratin-clumps surrounded by multilayered epithelial cells in culture. CONCLUSION: Krtap11-1 may play an important role on keratin-bundle assembly in the hair cortex and this study provides insight into the physical properties of the hair shaft.

S1-1/RBM10: multiplicity and cooperativity of nuclear localisation domains.

BACKGROUND INFORMATION: S1-1, also called RBM10, is an RNA-binding protein of 852 residues. An alteration of its activity causes TARP syndrome, a severe X-linked disorder with pre- or post-natal lethality in affected males. Its molecular function, although still largely unknown, has been suggested to be transcription and alternative splicing. In fact, S1-1 localises in the nucleus in tissue cells and cultured cells. RESULTS: By deletion and substitution mutagenesis, a classical 17-amino-acid (aa) nuclear localisation sequence (NLS1) was identified at aa 743-759 in the C-terminal region of S1-1. NLS1 was bipartite, with its N-terminal basic cluster weakly contributing to the NLS activity. S1-1 contained two additional NLSs. One was in the aa 60-136 RNA recognition motif region (NLS2), and the other was a novel NLS motif sequence in the aa 481-540 octamer-repeat (OCRE) region (NLS3). The OCRE is a domain known to be critical in splicing regulation, as shown with RBM5, a close homologue of RBM10 [Bonnal et al. (2008) Mol. Cell 32, 81-95]. The NLS activities were verified by expressing each DNA sequence linked to EGFP or a FLAG tag. These multiple NLSs acted cooperatively, and S1-1 became completely cytoplasmic after the concomitant removal of all NLS domains. In some cell types, however, S1-1 was partly cytoplasmic, suggesting that cellular localisation of S1-1 is subjected to regulation. CONCLUSIONS: The present results indicate that S1-1 contains multiple NLSs that act cooperatively. Among them, the OCRE is a hitherto unreported NLS. The nuclear localisation of S1-1 appears to be regulated under certain circumstances. We discuss these NLSs in relation to the biochemical processes they are involved in.

Incremental increases in glucocorticoid doses may reduce the risk of osmotic demyelination syndrome in a patient with hyponatremia due to central adrenal insufficiency.

A 50-year-old man was admitted to our hospital because of general malaise. Laboratory tests revealed severe hyponatremia (104 mEq/L), which was attributed to central adrenal insufficiency. To treat presumed central diabetes insipidus (CDI), we administered a small dose of hydrocortisone and gradually increased it to maintenance doses to prevent osmotic demyelination syndrome (ODS). Serum sodium levels did not increase more than 10 mEq/L/day and ODS did not occur. Thereafter, the patient was proven to have CDI. Incremental increases in glucocorticoid dose may reduce the risk of ODS for patients with hyponatremia due to central adrenal insufficiency, especially that complicated by CDI.

Onset of adult-onset Still's disease following influenza vaccination.

We describe that case of a 61-year-old woman who developed high spiking fever, sore throat, polyarthralgia, and salmon pink evanescent rash following influenza vaccination. A diagnosis of adult-onset Still's disease (AOSD) was made based on clinical and laboratory findings. Methylprednisolone pulse therapy followed by oral prednisolone resulted in a favorable outcome. This is the second published case in which a causal relationship between vaccination and onset of AOSD is suggested. Bystander activation would appear to play an important role in inducing the immune reaction.

Meta-analysis of laparoscopy-assisted and open distal gastrectomy for gastric cancer.

BACKGROUND: Laparoscopy-assisted distal gastrectomy (LADG) for gastric cancer is a minimally invasive technique. We performed a meta-analysis of five randomized clinical trials (RCTs) to evaluate and compare the benefits of LADG with those of open distal gastrectomy (ODG). METHODS: The present meta-analysis was based on the comparison of LADG with ODG for gastric cancer. The following factors were examined: operative time, estimated blood loss, number of harvested lymph nodes, time to resumption of oral intake, duration of hospital stay, frequency of analgesic administration, complications, tumor recurrence, and mortality. RESULTS: We selected five RCTs to compare LADG with ODG for gastric cancer. A total of 326 patients with gastric cancer were included in this meta-analysis of whom 164 underwent LADG and 162 underwent ODG. There was a significant difference in the volume of intraoperative blood loss, period of hospital stay, frequency of analgesic administration, and rate of complications between LADG and ODG. There was no difference in the resumption of oral intake, rate of tumor recurrence, and mortality. The operative time was significantly longer and the number of harvested lymph nodes was significantly smaller in LADG than in ODG. CONCLUSION: LADG is significantly superior to ODG regarding the volume of blood loss, duration of hospital stay, level of pain, and risk of complications. There was no difference in the resumption of oral intake, rate of tumor recurrence, and mortality. However, LADG was significantly inferior to ODG regarding operative time and also had a smaller number of harvested lymph nodes.

A meta-analysis of randomized controlled trials that compared laparoscopy-assisted and open distal gastrectomy for early gastric cancer.

BACKGROUND: We conducted a meta-analysis to evaluate and compare the advantages of laparoscopy-assisted distal gastrectomy (LADG) over open distal gastrectomy (ODG) for treating early gastric cancer (EGC). METHODS: We searched MEDLINE, EMBASE, Science Citation Index, and Cochrane Controlled Trial Register for relevant papers published between January 1990 and January 2010 by using the following search terms: laparoscopy-assisted gastrectomy, laparoscopic gastrectomy, and early gastric cancer. The following data were analyzed: operative time, estimated blood loss, number of harvested lymph nodes, time required for resumption of oral intake, duration of hospital stay, frequency of analgesic administration, complications, tumor recurrence, and mortality. RESULTS: We selected four papers reporting randomized control studies (RCTs) that compared LADG with ODG for EGC. Our meta-analysis included 267 patients with EGC; of these, 134 and 133 had undergone LADG and ODG, respectively. The volume of intraoperative blood loss, frequency of analgesic administration, and rate of complications were significantly lesser for LADG than for ODG. However, the time required for resumption of oral intake and duration of hospital stay did not significantly differ between LADG and ODG. The operative time for LADG was significantly longer than that for ODG; further, the number of harvested lymph nodes was significantly lesser in the LADG group than in the ODG group. CONCLUSION: LADG is advantageous over ODG because it results in lesser blood loss, is less painful, and is associated with a low risk of complications. Additional RCTs that compare LADG and ODG and investigate the long-term oncological outcomes of LADG are required to determine the advantages of LADG over ODG.

Lamivudine therapy for hepatitis B virus reactivation in a patient receiving intra-arterial chemotherapy for advanced hepatocellular carcinoma.

We describe a case of hepatitis B virus (HBV) reactivation that responded to lamivudine therapy in a 58-year-old man with advanced hepatocellular carcinoma (HCC). After seroconversion of hepatitis B e antigen to e antibodies by interferon therapy, the patient was found to have HCC with a portal tumor thrombus. A transarterial port was placed in the right femoral artery to permit infusion of epirubicin and cisplatin. After 3 months of arterial chemotherapy, the serum alpha-fetoprotein level had decreased and tumor staining diminished. Laboratory examinations suggested a flare-up of hepatitis B. Lamivudine was given to manage HBV reactivation. After 1 month, the serum HBV DNA level fell below the detection limit, and the alanine aminotransferase activity decreased to the normal range. With further arterial chemotherapy for HCC, no tumor staining was detected on computed tomography. Administration of lamivudine decreased serum HBV DNA levels for 7 months. Our findings suggest that HBV may be reactivated during chemotherapy for HCC, similar to other types of malignancies, and that lamivudine is effective for the management of HBV reactivation.

Erythrocyte-binding polyamine as a tumor growth marker for human hepatocellular carcinoma.

BACKGROUND/AIMS: Polyamines are essential for cell proliferation, differentiation, and transformation. Concentrations of polyamines are higher in some cancer tissue than in normal tissue. We examined erythrocyte-binding polyamines to evaluate the usefulness of polyamines as pathophysiological markers for hepatocellular carcinoma. METHODOLOGY: We measured erythrocyte-binding polyamine levels in peripheral blood samples obtained from 51 normal adult controls, 136 patients with chronic viral hepatitis, 104 patients with viral hepatic cirrhosis, and 130 patients with hepatocellular carcinoma. RESULTS: We defined the concentration of spermidine plus spermine as the erythrocyte polyamine level, and designated the cut-off level for normal as the mean erythrocyte polyamine level +/- 2 SD in control. The erythrocyte polyamine level was abnormally elevated (positive) in 56 (43%) of 130 patients with hepatocellular carcinoma, 11 (8%) of 136 patients with chronic hepatitis, and 13 (13%) of 108 patients with cirrhosis. The level was higher in patients with a short tumor doubling time. In 27 patients with tumors, there was negative correlation between tumor doubling time and erythrocyte polyamine level (r = -0.46; P = 0.0147). CONCLUSIONS: We conclude that erythrocyte polyamine may be a useful tumor growth marker in patients with hepatocellular carcinoma.