Wide-Targeted Semi-Quantitative Analysis of Acidic Glycosphingolipids in Cell Lines and Urine to Develop Potential Screening Biomarkers for Renal Cell Carcinoma.

Glycosphingolipids (GSLs), mainly located in the cell membrane, play various roles in cancer cell function. GSLs have potential as renal cell carcinoma (RCC) biomarkers; however, their analysis in body fluids is challenging because of the complexity of numerous glycans and ceramides. Therefore, we applied wide-targeted lipidomics using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with selected reaction monitoring (SRM) based on theoretical mass to perform a comprehensive measurement of GSLs and evaluate their potency as urinary biomarkers. In semi-quantitative lipidomics, 240 SRM transitions were set based on the reported/speculated structures. We verified the feasibility of measuring GSLs in cells and medium and found that disialosyl globopentaosylceramide (DSGb5 (d18:1/16:0)) increased GSL in the ACHN medium. LC-MS/MS analysis of urine samples from clear cell RCC (ccRCC) patients and healthy controls showed a significant increase in the peak intensity of urinary DSGb5 (d18:1/16:0) in the ccRCC group compared with that in the control group. Receiver operating characteristic analysis indicated that urinary DSGb5 could serve as a sensitive and specific marker for RCC screening, with an AUC of 0.89. This study demonstrated the possibility of urinary screening using DSGb5 (d18:1/16:0). In conclusion, urinary DSGb5 (d18:1/16:0) was a potential biomarker for cancer screening, which could contribute to the treatment of RCC patients.

Systemic Immune-Inflammation Index Predicts Tumor Recurrence after Radical Resection for Colorectal Cancer.

The systemic inflammatory response is associated with tumor promotion and suppression. Accumulating evidence shows that peripheral blood markers of inflammatory response predict clinical outcomes in various human cancers. The aim of this study was to investigate the prognostic relevance of the inflammation-based biomarkers in colorectal cancer (CRC). We retrospectively analyzed 118 CRC patients who underwent curative resection between 2012 and 2017. The inflammation-based biomarkers were evaluated by using preoperative neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), systemic immune-inflammation index (SII), systemic inflammation response index (SIRI), prognostic nutritional index (PNI), and Glasgow prognostic score (GPS). Prognostic values were assessed by the Kaplan-Meier analysis for cancer-specific recurrence-free survival (RFS) and Cox proportional-hazards model. There were significant differences in the levels of NLR, PLR, SII, and SIRI between recurrence and non-recurrence group. The area under the curve (AUC) for SII was 0.710, which showed the highest value in the inflammation-based biomarkers. Multivariate analysis identified that SII (p = 0.0031) and lymph node metastasis (p = 0.0168) were independent prognostic factors for recurrence. High SII exhibited more dismal RFS than low SII in CRC patients with non-metastatic lymph node (p = 0.0002). Our study suggests that SII and lymph node metastasis could be useful indicators in predicting the recurrence of CRC patients. Additionally, SII could accurately stratify CRC patients with tumor recurrence by combining with lymph node metastasis. This result would be beneficial for determining the optimal therapeutic strategies after surgical resection for CRC.

The impact of gender difference on the relationship between serum high-density lipoprotein level and cardiovascular events in incident dialysis patients: a multicenter prospective cohort study.

PURPOSE: This study aimed to clarify the relationship between serum high-density lipoprotein cholesterol (HDL-C) level and incidence of CV events. Moreover, the relationship stratified by gender was evaluated. METHODS: The study database was derived from a multicenter, prospective, cohort study that included 1520 incident dialysis patients in Aichi Prefecture, Japan. Baseline was defined as the time of dialysis initiation and outcome as incidence of CV event. The enrolled patients were first classified into four quartile groups (Q1, Q2, Q3, and Q4; Q1 was the lowest serum HDL-C) based on serum HDL-C level. A multivariate Cox proportional hazards analysis was conducted using the stepwise regression method to determine factors associated with outcomes and was performed in each gender. RESULTS: The log-rank test revealed significant differences in the incidence of CV events between not only the four groups (p = 0.003) but also the male and female patients (HR, 0.74; 95% CI, 0.59-0.92; p < 0.001). Stepwise regression analysis indicated that higher serum HDL-C level was significantly associated with lower incidence of CV events (HR, 0.88; 95% CI, 0.82-0.95; p = 0.001). The association between serum HDL-C and incidence of CV events could be clarified in only female patients (male patients; HR, 0.92; 95% CI, 0.84-1.01; p = 0.067 and female patients; HR, 0.81; 95% CI, 0.72-0.92; p = 0.001). CONCLUSIONS: HDL-C level at the initiation of dialysis was associated with the incidence of CV events. Moreover, the association was prominent in female patients. TRIAL REGISTRATION: The trial registration number of the study is UMIN 7096, registered on January 18, 2012.

PLOD2 Is Essential to Functional Activation of Integrin ß1 for Invasion/Metastasis in Head and Neck Squamous Cell Carcinomas.

Identifying the specific functional regulator of integrin family molecules in cancer cells is critical because they are directly involved in tumor invasion and metastasis. Here we report high expression of PLOD2 in oropharyngeal squamous cell carcinomas (SCCs) and its critical role as a stabilizer of integrin ß1, enabling integrin ß1 to initiate tumor invasion/metastasis. Integrin ß1 stabilized by PLOD2-mediated hydroxylation was recruited to the plasma membrane, its functional site, and accelerated tumor cell motility, leading to tumor metastasis in vivo, whereas loss of PLOD2 expression abrogated it. In accordance with molecular analysis, examination of oropharyngeal SCC tissues from patients corroborated PLOD2 expression associated with integrin ß1 at the invasive front of tumor nests. PLOD2 is thus implicated as the key regulator of integrin ß1 that prominently regulates tumor invasion and metastasis, and it provides important clues engendering novel therapeutics for these intractable cancers.

Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae.

BACKGROUND: Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galß1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcß1-3GalNAcα1-O-Ser/Thr) has not been constructed. METHODS: To generate core 3-producing strain, genes encoding uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter, UDP-GlcNAc transporter, and two glycosyltransferases were integrated into the genome. A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. The amount of the resulting modified peptide was analyzed by HPLC. RESULTS: Introduction of a codon-optimized UDP-GlcNAc:ßGal ß-1,3-N-acetylglucosaminyltransferase 6 (ß3Gn-T6) gene yielded increases in ß3Gn-T6 activity but did not alter the level of core 3 production. The highest in vitro activity of ß3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. CONCLUSIONS: Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. Mn(2+) supplementation led to up-regulation of reaction of glycosylation in the Golgi, resulting in increases of core 3 production. GENERAL SIGNIFICANCE: This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae.

Proteinuria rebound in IgA nephropathy associated with obesity-related glomerulopathy.

IgA nephropathy (IgAN), the most prevalent primary chronic glomerulonephritis worldwide, has three major risk factors: hypertension, proteinuria >1 g/day, and severe renal lesions. Obesity also portends a poor prognosis. A Japanese boy with IgAN showed nephrotic syndrome at presentation. Pathological features resembled those of membranoproliferative glomerulonephritis (MPGN), although IgA deposition differed from MPGN and IgAN. Combination therapy improved renal lesions, but rebound deterioration of proteinuria occurred in this patient, who had marked obesity and hypertension. Serial kidney biopsy specimens were compatible with obesity-related glomerulopathy (ORG). Rebound proteinuria was apparently attributable to ORG rather than relapse and flaring up of IgAN.

Pulmonary veno-occlusive disease in an 11-year-old girl: diagnostic pitfalls.

Pulmonary veno-occlusive disease (PVOD) is a rare chronic lung disease that is difficult to diagnose due to non-specific clinical findings. Little is known about the pathogenesis of PVOD. Reported herein is the case of an 11-year-old girl who initially presented with 'bat-wing' shadows on chest radiography. This finding, coupled with prominent hemosiderosis in bronchoalveolar lavage fluid, initially led to a misdiagnosis of idiopathic pulmonary hemosiderosis. Oral prednisolone dramatically improved signs and symptoms initially, but her condition then gradually deteriorated during maintenance therapy with corticosteroids and other immunosuppressants. PVOD was suspected but not confirmed owing to a lack of hallmark radiographic findings and contraindications for lung biopsy. Three years later, while arranging for lung transplantation, the patient experienced sudden onset of fatal massive pulmonary edema. PVOD was confirmed at autopsy. This case provides insights regarding an unfamiliar presentation of PVOD and may help physicians to avoid diagnostic pitfalls.

Complete remission within 2 years predicts a good prognosis after methylprednisolone pulse therapy in patients with IgA nephropathy.

BACKGROUND: Pozzi et al. reported the effectiveness of steroid pulse therapy (Pozzi's regimen) in IgA nephropathy (IgAN). The present study was performed to clarify the predictive factors for IgAN patients treated with Pozzi's regimen. METHODS: One hundred nine IgAN patients treated by Pozzi's regimen were observed for up to 112.6 (median 39.7) months, and remission of proteinuria (PR) and disappearance of urinary abnormalities [complete remission (CR)] after Pozzi's regimen were analyzed. Predictive factors for the glomerular filtration rate (GFR) slopes for up to 5 years were analyzed among 81 patients who were observed for at least 2 years. The outcome of a 50 % increase in sCr was compared between the CR and non-CR groups within 2 years. RESULTS: Cumulative PR and CR rates increased rapidly until 2 years (54.5 and 46.8 % at 2 years), and then slowly but steadily up to 6 years (72.8 and 66.4 % at 6 years). Baseline characteristics of the CR and non-CR groups within 2 years were similar except for proteinuria. GFR slope was steeper in the non-CR group than in the CR group (-2.44 ± 5.12 vs. -0.32 ± 3.34 ml/min/1.73 m(2)/year). On multivariate analysis, sex and CR within 2 years were associated with GFR slope. Kaplan-Meier analysis demonstrated a better survival rate in CR group patients without a 50 % increase in sCr (p = 0.024). CONCLUSIONS: Among IgAN patients treated with Pozzi's regimen, CR within 2 years predicts a good prognosis.

Chemical synthesis of homogeneous human glycosyl-interferon-ß that exhibits potent antitumor activity in vivo.

Chemical synthesis of homogeneous human glycoproteins exhibiting bioactivity in vivo has been a challenging task. In an effort to overcome this long-standing problem, we selected interferon-ß and examined its synthesis. The 166 residue polypeptide chain of interferon-ß was prepared by covalent condensation of two synthetic peptide segments and a glycosylated synthetic peptide bearing a complex-type glycan of biological origin. The peptides were covalently condensed by native chemical ligation. Selective desulfurization followed by deprotection of the two Cys(Acm) residues gave the target full-length polypeptide chain of interferon-ß bearing either a complex-type sialyl biantennary oligosaccharide or its asialo form. Subsequent folding with concomitant formation of the native disulfide bond afforded correctly folded homogeneous glycosyl-interferon-ß. The chemically synthesized sialyl interferon-ß exhibited potent antitumor activity in vivo.

Chlamydia pneumoniae infection-related hemophagocytic lymphohistiocytosis and acute encephalitis and poliomyelitis-like flaccid paralysis.

A 3-year-old male presented with Chlamydia pneumoniae infection-related hemophagocytic lymphohistiocytosis (HLH). The patient developed an episode of HLH with severe skin eruption following C. pneumoniae pneumonia. Symptoms responded to steroid/cyclosporine A therapy, but the patient slowly lost consciousness and developed systemic flaccid paralysis. He was diagnosed with encephalitis/myelitis by brain and spinal MRI. Neurological symptoms and signs gradually resolved. We thought that the immune response to C. pneumoniae infection triggered the development of HLH, associated with unusual neurological complications. This report describes a novel case of C. pneumoniae-associated HLH and with poliomyelitis like flaccid paralysis.

Lymphatic vessels develop during tubulointerstitial fibrosis.

Recent progress with specific markers of lymphatic vessel endothelium allowed recognition of lymphangiogenic events in various disease states; however, there is little information concerning this process in human chronic renal diseases. To determine this we measured expression of the lymphatic marker D2-40 and vascular endothelial growth factor-C (VEGF-C), a major growth factor in lymphangiogenesis, in 124 human renal biopsy specimens. In the kidneys of control subjects and in uninjured areas of pathologic specimens, lymphatic vessels were detected only around the arcuate and interlobular arteries. An increase in the number of lymphatic vessels was found at the site of tubulointerstitial lesions correlating with the degree of tissue damage and more strongly correlating with areas of fibrosis than inflammation. On serial sections, lymphatic vessel proliferation was found in the tubulointerstitial area at the site of tuft adhesions to Bowman's capsule. Lymphatic growth was associated with VEGF-C expression in inflammatory mononuclear cells and tubular epithelial cells, mainly of proximal tubules. Lymphangiogenesis and VEGF-C expression was elevated in diabetic nephropathy in comparison to other renal diseases. Our results indicate that lymphangiogenesis is a common feature in the progression of the tubulointerstitial fibrosis.

HIV prevention risks for Black women in Canada.

The future availability of HIV vaccines can increase options available to Canadian Black women for risk reduction. However, current conceptual frameworks do not adequately address barriers to HIV prevention for this population, and may be inadequate to address challenges with vaccines. This study explored knowledge and attitudes regarding HIV vaccines and associated prevention methods to inform appropriate conceptual frameworks for their dissemination to Canadian Black women. We completed four 90-min focus groups with women (n=26) of African or Caribbean origins, and six interviews with key informants providing health and social services in the Black communities of Toronto. The participants suggested that there were significant risks associated with seeking prevention information and attempting to reduce exposure to HIV infection. They described individual, familial, community and institutional domains of risk and predicted the same spectrum of risk for HIV vaccines. Participants advocated for education, empowerment and institutional change to create a supportive environment for vaccines and other HIV prevention methods. They further indicated that preparation for vaccine dissemination will need to prioritize building trust between women of the Black communities and institutions in the research, health and government sectors.

How do organizations and social policies 'acculturate' to immigrants? Accommodating skilled immigrants in Canada.

While the idea of acculturation (Berry 1997) was originally proposed as the mutual change of both parties (e.g., immigrants and the host society), the change processes of host societies are neglected in research. A grounded theory study explored the efforts of human service organizations to 'acculturate' to an increasingly diverse immigrant population, through interviews conducted with service providers serving Mainland Chinese immigrants. Acculturation efforts of human service organizations (mezzo-level acculturation) were often needs-driven and affected by the political will and resultant funding programs (macro-level forces). Even with limitations, human service organizations commonly focused on hiring Mainland Chinese immigrants to reflect the changing demographics of their clientele and creating new programs to meet the language and cultural backgrounds of the clients. To contextualize these organizational efforts, an analysis of how policy changes (macro-level acculturation) interact with organizational practice is presented. Finally, the meaning of acculturation for the host society is discussed.

High mobility group box chromosomal protein 1 in patients with renal diseases.

BACKGROUND/AIM: The high mobility group box chromosomal protein 1 (HMGB1), a nuclear DNA-binding protein, has recently been recognized as a new proinflammatory cytokine. The purpose of this study was to examine the significance of HMGB1 in patients with renal diseases. METHODS: HMGB1 concentrations in sera were measured by enzyme-linked immunosorbent assay, and antibodies against HMGB1 were examined by Western blotting in patients who underwent renal biopsies and in healthy controls. Immunohistochemistry for HMGB1 was also performed. RESULTS: Serum HMGB1 was more likely to be positive in patients who underwent renal biopsies as compared with the controls. Patients with anti-neutrophil cytoplasmic antibody-related glomerulonephritis (ANCA-GN) and those with Henoch-Schonlein purpura nephritis showed a significantly higher tendency to be HMGB1 positive. The presence of anti-HMGB1 antibody was not associated with the presence of serum HMGB1. Immunohistochemistry revealed that HMGB1 was expressed in mononuclear cells in the interstitium or in the glomeruli of some patients with ANCA-GN or IgA nephropathy (IgAN). Subanalysis demonstrated that among patients with IgAN, those who had crescent formation showed a higher tendency to be HMGB1 positive than those who did not. CONCLUSIONS: HMGB1 was expressed in the sera of patients with renal diseases who underwent renal biopsies, especially among those who had vasculitis including ANCA-GN, Henoch-Schonlein purpura nephritis, and IgAN with glomerular crescents.

When family enters the picture: the model of cultural negotiation and gendered experiences of Japanese academic sojourners in the United States.

A grounded-theory study aimed at reconceptualizing cultural adaptation processes from gender role and family/couple perspectives while critically drawing from acculturation and culture and self literatures. In-depth interviews with 34 Japanese academic sojourners (international students, scholars) and their spouses (a total of 50 interviews with select longitudinal interviews) were conducted. The author earlier developed the Model of Cultural Negotiation (2001; 2006) capturing uneven and cyclical processes of dealing with multiple cultural contexts. The current study further develops more tailored versions of this model, Family-Based (Couple-Based) Cultural Negotiation and Individual-Based Cultural Negotiation, highlighting the impacts of family/couple and gender roles, especially for female spouses. These conceptualizations afford a sophisticated understanding of the processes of culture.

2-(1,3-Dioxan-2-yl)ethylsulfonyl group: a new versatile protecting and activating group for amine synthesis.

[reaction: see text] 2-(1,3-Dioxan-2-yl)ethylsulfonyl (Dios) chloride was synthesized and used as a new versatile sulfonating agent for amines. Primary and secondary amines were sulfonated very easily in excellent yields with Dios chloride. N-Nonsubstituted and N-monosubstituted Dios-amides, activated amines, were alkylated satisfactorily under new Mitsunobu conditions utilizing (cyanomethylene)tributylphosphorane (CMBP). The Dios group is very stable under basic and reductive conditions and is removed by heating in a hot aqueous solution of trifluoroacetic acid.

The role of critical consciousness in multicultural practice: examining how its strength becomes its limitation.

Given the strengths of developing critical consciousness in multicultural practice, why do roadblocks persist? This article examines how the strengths of developing critical consciousness in multicultural practice can paradoxically become its limitation. Literatures from counseling psychology, clinical psychology, social psychology, social work, feminist theory, and critical theory are integrated in a discussion of the various components and strengths involved in working toward critical consciousness. This literature is then used to discuss some of the cognitive and affective limitations to achieving critical consciousness. Implications for practice are discussed.

Preparation of (Cyanomethylene)tributylphosphorane: a new mitsunobu-type reagent.

(Cyanomethylene)tributylphosphorane (CMBP), which promoted the alkylation of various nucleophiles (HA) with alcohols (ROH) to give RA (Mitsunobu-type reaction), was prepared in two steps starting from chloroacetonitrile.