Prostate-specific membrane antigen can promote in vivo osseous metastasis of prostate cancer cells in mice

Reports remain insufficient on whether and how prostate-specific membrane antigen (PSMA) can influence in vivo osseous metastasis of prostate cancer (PCa). In the present study, the authors induced stable expression of PSMA in mouse PCa cell line RM-1. In vivo osseous metastasis was induced in 37 6-week-old female C57BL/6 mice weighing 22.45 ± 0.456 g. RM-1 cells were actively injected into the femoral bone cavity, leading to bilateral dissymmetry of bone density in the femoral bone. Tumor cells were also detected in bone tissue by pathological examination. The impact on bone density was demonstrated by the significant difference between animals injected with RM-PSMA cells (0.0738 ± 0.0185 g/cm²) and animals injected with RM-empty plasmid cells (0.0895 ± 0.0241 g/cm²). The lytic bone lesion of the RM-PSMA group (68.4%) was higher than that of the control group (27.8%). Immunohistochemistry showed that the expression of both vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) was distinctly higher in the RM-PSMA group than in the control group, while ELISA and Western blot assay indicated that VEGF and MMP-9 were higher in the RM-PSMA group compared to the control group (in vitro). Thus, the present study proposed and then confirmed for the first time that PSMA can promote in vivo osseous metastasis of PCa by increasing sclerotic destruction of PCa cells. Further analyses also suggested that PSMA functions positively on the invasive ability of RM-1 by increasing the expression of MMP-9 and VEGF by osseous metastases in vivo

RNAi-mediated knockdown of pituitary tumor- transforming gene-1 (PTTG1) suppresses the proliferation and invasive potential of PC3 human prostate cancer cells

Pituitary tumor-transforming gene-1 (PTTG1) is a proto-oncogene that promotes tumorigenesis and metastasis in numerous cell types and is overexpressed in a variety of human tumors. We have demonstrated that PTTG1 expression was up-regulated in both human prostate cancer specimens and prostate cancer cell lines. For a more direct assessment of the function of PTTG1 in prostate tumorigenesis, RNAi-mediated knockdown was used to selectively decrease PTTG1 expression in PC3 human prostate tumor cells. After three weeks of selection, colonies stably transfected with PTTG1-targeted RNAi (the knockdown PC3 cell line) or empty vector (the control PC3 cell line) were selected and expanded to investigate the role of PTTG1 expression in PC3 cell growth and invasion. Cell proliferation rate was significantly slower (28%) in the PTTG1 knockdown line after 6 days of growth as indicated by an MTT cell viability assay (P < 0.05). Similarly, a soft agar colony formation assay revealed significantly fewer (66.7%) PTTG1 knockdown PC3 cell colonies than control colonies after three weeks of growth. In addition, PTTG1 knockdown resulted in cell cycle arrest at G1 as indicated by fluorescence-activated cell sorting. The PTTG1 knockdown PC3 cell line also exhibited significantly reduced migration through Matrigel in a transwell assay of invasive potential, and down-regulation of PTTG1 could lead to increased sensitivity of these prostate cancer cells to a commonly used anticancer drug, taxol. Thus, PTTG1 expression is crucial for PC3 cell proliferation and invasion, and could be a promising new target for prostate cancer therapy.

Delivery of the Sox9 gene promotes chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells in an in vitro model

SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.

Relationship between salt consumption measured by 24-h urine collection and blood pressure in the adult population of Vitória (Brazil)

High salt intake is related to an increase in blood pressure and development of hypertension. However, currently, there are no national representative data in Brazil using the gold standard method of 24-h urine collection to measure sodium consumption. This study aimed to determine salt intake based on 24-h urine collection in a sample of 272 adults of both genders and to correlate it with blood pressure levels. We used a rigorous protocol to assure an empty bladder prior to initiating urine collection. We excluded subjects with a urine volume <500 mL, collection period outside of an interval of 23-25 h, and subjects with creatinine excretion that was not within the range of 14.4-33.6 mg/kg (men) and 10.8-25.2 mg/kg (women). The mean salt intake was 10.4±4.1 g/day (d), and 94% of the participants (98% of men and 90% of women) ingested more than the recommended level of 5 g/d. We found a positive association between salt and body mass index (BMI) categories, as well as with salt and blood pressure, independent of age and BMI. The difference in systolic blood pressure reached 13 mmHg between subjects consuming less than 6 g/d of salt and those ingesting more than 18 g/d. Subjects with hypertension had a higher estimated salt intake than normotensive subjects (11.4±5.0 vs 9.8±3.6 g/d, P<0.01), regardless of whether they were under treatment. Our data indicate the need for interventions to reduce sodium intake, as well the need for ongoing, appropriate monitoring of salt consumption in the general population.

Nefrolitíase em pacientes pediátricos: investigação metabólica e anatômica

Desordens metabólicas são frequentes em pacientes com nefrolitíase pediátrica. OBJETIVOS: Estudar as alterações metabólicas e anatômicas e a análise química dos cálculos encontrados em pacientes da nossa região. MÉTODOS: Este é um estudo retrospectivo em 158 crianças com evidência de formação recente de cálculos, destes apenas 109 concluíram a investigação metabólica. A investigação laboratorial consistiu de duas amostras de urina de 24 horas com dosagem de cálcio, ácido úrico, citrato, oxalato, sódio e creatinina, cistinúria qualitativa, pH urinário seguido de 12 horas de jejum e restrição hídrica, cultura da urina e análise química quando os cálculos foram disponíveis. As técnicas de imagem incluíram ultrassonografia do trato urinário e urografia excretora. RESULTADOS: Em 96,3% das crianças alguma causa foi detectada. A principal alteração metabólica encontrada foi a hipercalciúria (73,4%). Análise química dos cálculos mostrou oxalato de cálcio em 90,9% dos casos. Alterações anatômicas foram encontradas em 18,0% dos pacientes investigados, e a mais frequente foi a duplicação pieloureteral (28,6%). CONCLUSÕES:Hipercalciúria foi a desordem mais encontrada, a alteração anatômica mais comum foi a duplicação pieloureteral e oxalato de cálcio foi o constituinte químico mais frequente. Este trabalho serviu para o conhecimento das características dos pacientes pediátricos portadores de nefrolitíase em nossa região.

Marcadores moleculares microssatélites na avaliação de sementes de soja com variação na coloração do hilo

Marcadores microssatélite têm sido utilizados na determinação da pureza genética em programas de melhoramento e de produção de sementes, visando garantir a qualidade genética das sementes. Objetivou-se neste trabalho demonstrar, com auxílio de marcadores moleculares microssatélites, que variações na coloração do hilo em sementes de soja, muitas vezes não constituem variação genética. Foram avaliadas sementes de soja da cultivar CD 222 que apresenta sementes com hilo preto, e de duas linhagens (CD 02RV-8444 e CD 01RV-7618), as quais apresentam sementes com hilo marrom. As sementes com variações na coloração e tonalidade do hilo encontradas em cada genótipo foram separadas visualmente em grupos. O DNA de cada semente foi extraído e analisado com marcadores microssatélites em "bulks" contendo DNA de cinco sementes. As variações encontradas na coloração do hilo da cultivar CD 222 e da linhagem CD 02RV-8444 não foram devidas à variação genética, considerando 16 locos microssatélites. Para a linhagem CD 01RV-7618, os bulks de sementes com hilo marrom pigmentado de preto (B1), com hilo marrom mais intenso (B2) e intermediário (B3), foram considerados iguais. Para esta mesma linhagem foi detectada diferença no bulk de sementes com hilo marrom menos intenso (B4), com o primer Satt 070. Duas sementes deste bulk apresentaram o mesmo padrão molecular observado para os outros bulks, no mesmo loco. As demais sementes foram consideradas misturas. A utilização de marcadores microssatélites demonstra que variações na cor ou tonalidade do hilo da soja nem sempre correspondem à variação genética.

Bulgakov's sophiology as philosopheme: non-ontology and ontogenesis

The text examines Sergej Nikolajeviè Bulgakov's description of the philosopheme as thoroughly "immanent" (viz., the immanence of man qua being, such that ontology in Bulgakov becomes a conceptual analogue for immanence) and the corollary that such immanence necessarily excludes the problematic of the "creation of the world." Because of this resolute immanence and the notion that the creation of the world in the form of creatio ex nihilo requires a non-immanent or non-ontological thought and concept, the problematic for Bulgakov is approached only by a theologeme. Appropriating this argument as material for a cursory philosopheme, the text attempts to transform Bulgakov's theologeme into a philosopheme through an elision of God and dogma that overdetermines the theologeme. This philosopheme (nascent within Bulgakov's work itself, in both his hesitation to the overdetermination of immanence and the commitment to the problem of creation) would be a thoroughly non-ontological philosopheme, one that allows for the treatment of the problematic of "creation" or singular ontogenesis, yet with the corollary that this philosopheme must rely on an "ontological zero" Such a philosopheme qua ontologically empty formula nevertheless remains ontologically significant insofar as it is to evince the limit of ontology, in the ontological zero's non-relationality to ontology.