FTY720 inhibits mesothelioma growth in vitro and in a syngeneic mouse model

Background: Malignant mesothelioma (MM) is a very aggressive type of cancer, with a dismal prognosis and inherent resistance to chemotherapeutics. Development and evaluation of new therapeutic approaches is highly needed. Immunosuppressant FTY720, approved for multiple sclerosis treatment, has recently raised attention for its anti-tumor activity in a variety of cancers. However, its therapeutic potential in MM has not been evaluated yet. Methods: Cell viability and anchorage-independent growth were evaluated in a panel of MM cell lines and human mesothelial cells (HM) upon FTY720 treatment to assess in vitro anti-tumor efficacy. The mechanism of action of FTY720 in MM was assessed by measuring the activity of phosphatase protein 2A (PP2A)-a major target of FTY720. The binding of the endogenous inhibitor SET to PP2A in presence of FTY720 was evaluated by immunoblotting and immunoprecipitation. Signaling and activation of programmed cell death were evaluated by immunoblotting and flow cytometry. A syngeneic mouse model was used to evaluate anti-tumor efficacy and toxicity profile of FTY720 in vivo. Results: We show that FTY720 significantly suppressed MM cell viability and anchorage-independent growth without affecting normal HM cells. FTY720 inhibited the phosphatase activity of PP2A by displacement of SET protein, which appeared overexpressed in MM, as compared to HM cells. FTY720 promoted AKT dephosphorylation and Bcl-2 degradation, leading to induction of programmed cell death, as demonstrated by caspase-3 and PARP activation, as well as by cytochrome c and AIF intracellular translocation. Moreover, FTY720 administration in vivo effectively reduced tumor burden in mice without apparent toxicity. Conclusions: Our preclinical data indicate that FTY720 is a potentially promising therapeutic agent for MM treatment

Studi Potensi Jumlah Penumpang Bus Pemadu Moda Rute Malang – Bandar Udara Juanda Pp

Bandar Udara Malang yang belum melayani banyak tujuan penerbangan membuat pengguna moda pesawat memilih Bandar Udara Juanda. Disisi lain angkutan yang melayani rute Malang-Juanda PP hanya angkutan travel. Untuk itu dibutuhkan moda lain yang lebih ekonomis dan memiliki kapasitas lebih banyak dibandingkan angkutan travel. Bus pemadu moda adalah moda alternatif yang dapat memenuhi kebutuhan tersebut.Pengumpulan data dilakukan dengan penyebaran kuisioner karakteristik sosial-ekonomi, karakteristik perjalanan serta kuisioner dengan teknik penyusunan stated preference. Stated preference memiliki atribut biaya perjalanan, waktu tempuh dan frekuensi keberangkatan. Sedangkan untuk prediksi tarif bus pemadu moda yang direncanakan diperoleh dari perhitungan BOK. Tarif yang telah diperoleh dari perhitungan BOK dibandingkan dengan nilai ATP dan WTP yang diperoleh dari kuisioner yang telah disebarkan. Sehingga didapatkan tarif ideal yang akan diberlakukan apabila bus pemadu moda tersebut direalisasikan.Setelah melakukan perhitungan tarif berdasarkan BOK diperoleh tarif sebesar Rp 23.374,- serta berdasarkan ATP dan WTP diperoleh tarif sebesar Rp 43.675,-. Dengan demikian perkiraan awal tarif bus pemadu moda sebesar Rp 40.000,- dapat diberlakukan. Hasil dari pemodelan pemilihan moda dengan metode stated preference untuk selisih biaya perjalanan Malang-Juanda: dan Juanda-Malang : , untuk selisih waktu tempuh perjalanan () rute Malang-Juanda : dan rute Juanda-Malang : , sedangkan untuk selisih Frekuensi Keberangkatan () rute Malang-Juanda : dan rute Juanda-Malang : .Potensi perpindahan pengguna travel ke bus pemadu moda rute Malang-Juanda sebanyak 705 orang per hari (83,97%). Sedangkan untuk rute Juanda-Malang sebanyak 1516 orang per hari (90,24%)

The role of chemerin in hypertension

Cząsteczki dwufunkcyjne, takie jak chemeryna, identyfikowane są jako łącznik pomiędzy dysfunkcjami metabolicznymi i odpowiedzią immunologiczną. Chemeryna pełni funkcję chemoatraktanta dla komórek immunologicznych takich jak pDC (plazmocytoidalne komórki dendrytyczne) oraz makrofagi, natomiast jako adipokina reguluje metabolizm adipocytów. Najnowsze badania wykazały zaangażowanie mechanizmów immunologicznych w rozwoju nadciśnienia, w którym znaczącą rolę przypisuje się również dysfunkcjom metabolicznym. Wobec tego, uzasadnione było zbadanie roli chemeryny w rozwoju tej choroby.Nadciśnienie tętnicze u myszy indukowane było poprzez infuzję ANGII (angiotensyny II). W pierwszym etapie porównano poziom ekspresji genu chemeryny w myszach kontrolnych oraz myszach z nadciśnieniem w PVAT (tkance tłuszczowej okołonaczyniowej), wątrobie oraz tłuszczu trzewnym. Wykazano podwyższoną ekspresję genu w PVAT. Powyższy wynik, potwierdzony został na poziomie białka poprzez ilościowy pomiar chemeryny obecnej w nadsączach z izolowanej PVAT. Przy pomocy barwień fluorescencyjnych określono dokładną lokalizację chemeryny w otoczeniu aort. Zarówno w przypadku skrawków kontrolnych jak i pochodzących z myszy z nadciśnieniem wykazano obecność chemeryny w PVAT. Najbogatszym jej źródłem były jednak ściany aort. Ponadto sygnał zidentyfikowano w mniejszych naczyniach krwionośnych oraz zwojach nerwowych. Średnia intensywność sygnału fluorescencji chemeryny wykazała istotne statystycznie podwyższenie w ścianach aort myszy z nadciśnieniem, korelujące z wynikiem pomiaru szerokości ścian aort. W kolejnym etapie badano potencjalną funkcję chemeryny jako chemoatraktanta dla komórek immunologicznych w PVAT. Zobrazowano nacieki makrofagów w obszarze okołoaortalnym myszy z nadciśnieniem. Nie wykazano jednak chemotaksji komórek transfekowanych receptorem dla chemeryny do nadsączów z PVAT oraz ścian aort.Podsumowując, praca dostarcza dowodów na korelację pomiędzy poziomem chemeryny a nadciśnieniem. Otrzymane wyniki nie dowodzą jednoznacznie chemotaktycznej roli chemeryny w tym modelu, sugerują jednak inne potencjalne funkcje tego białka w nadciśnieniu.Bifunctional molecules such as chemerin, have been indentified as the link between metabolic disorders and immune responses. Chemerin modulates immunity acting as chemoattractant for pDC (plasmocytoid dendritic cells) and macrophages, while as an adipokine it may regulate adipose tissue metabolism. Growing number of studies support a role of immune factor in hypertension, a disease associated with metabolic disorders. Thus the objective of this study was to investigate the contribution of chemerin to the hypertension.Hypertension was induced in mice by infusion of AGNII (angiotensin II). First, we compared chemerin expression levels in hypertensive and control mice and showed increased chemerin expression in PVAT of hypertensive mice. Increase at mRNA level was in line with upregulated chemerin levels in supernatants from isolated PVAT. In contrast to the PVAT, liver and visceral fat contained similar amount of chemerin mRNAs in untreated and AGNII-treated mice. Immunofluorescence staining provided an insight into the localization of chemerin in the aortic milieu. In both, hypertensive and control sections, chemerin was present in PVAT, however aortic walls appeared to be the most abundant source of chemerin. Mean fluorescence intensity confirmed significant upregulation of chemerin levels in the aortas of hypertensive mice that correlated with aortic walls thickness. Chemerin was also immunodetected in ganglia and accompanying vessels. In the next step, potential chemotactic role of chemerin in hypertension was tested. Chemerin chemotactic target-macrophages was found to accumulate in hypertensive PVAT. However, no migration of cells transfected with chemerin receptor towards PVAT and aortic walls supernatants was found in chemotaxis tests in vitro.In conclusion, correlation between chemerin levels and hypertension, but not clear chemotactic role of chemerin was demonstrated in this study. These data have broadened our perspective on possible chemerin functions in hypertension

Sage-modified polydimethylsiloxane applied as antibacterial wound dressing material

Materials based on polydimethylsiloxane have been developed with antibacterial activity and mechanical properties required for wound treatment. Sage herb (raw, modified, and polyphenol extract) was used as afiller in an amount of 5 and 10% by weight. Physicochemical, mechanical, and biological properties were examined. The results indicate abeneficial effect of sage modification on the tested properties

Association of RAD51C germline mutations with breast cancer among Bahamians

RAD51C is known as an ovarian cancer gene; however, its role in breast cancer susceptibility is less clear. As part of a larger study, we assessed the role of germline RAD51C mutations in breast cancer development. We studied 387 unselected, BRCA1- and BRCA2-negative, Bahamian breast cancer cases and 653 controls to search for novel genetic associations with breast cancer development. During the first phase of the study, whole exome sequencing was utilized in 96 cases to identify an association between novel genes and breast cancer susceptibility. In the second phase of the study, targeted gene sequencing was utilized in the entirety of the cases and controls to identify an association between novel genetic mutations and breast cancer development. A RAD51C mutation was found in five breast cancer cases and in no control (5/387 versus 0/653; p = 0.007). None of the mutation-positive cases reported a family history of ovarian cancer. These data support increasing evidence that RAD51C mutations contribute to breast cancer susceptibility, although the impact may vary substantially from country to country

A Single nucleotide polymorphism in the ALDH2 gene modifies the risk of esophageal squamous cell carcinoma in BRCA2 p.K3326* carriers.

Esophageal squamous cell carcinoma (ESCC) has a very high incidence rate in northeastern Iran. Our team previously reported the BReast CAncer gene 2 (BRCA2) p.K3326* mutation as a moderately penetrant ESCC susceptibility variant in northern Iran (odds ratio (OR) = 3.64, 95% confidence interval (CI) = 1.74-7.59, P = 0.0003). Recently, it has been reported that aldehydes can induce BRCA2 haploinsufficiency in cells with a heterozygous pathogenic BRCA2 mutation and predispose them to carcinogenic effects. Based on this observation, we speculate that dysfunctional variants in Aldehyde Dehydrogenase 2 Family Member (ALDH2) may result in aldehyde-induced BRCA2 haploinsufficiency and increase cancer risk in BRCA2 mutation carriers. In support of this hypothesis, our team recently reported the breast cancer risk modifying effect of an ALDH2 common polymorphism, rs10744777, among Polish carriers of the BRCA2 p.K3326* mutation. In the current case-control study, we aimed to investigate the ESCC risk modifying effect of this ALDH2 polymorphism among BRCA2 p.K3326* mutation carriers. We assessed the interaction between the ALDH2 rs10744777 polymorphism and BRCA2 p.K3326* mutation in ESCC risk by genotyping this ALDH2 variant in the germline DNA of 746 ESCC cases and 1,373 controls from northern Iran who were previously genotyped for the BRCA2 p.K3326* mutation. Among a total of 464 individuals with TT genotype of the ALDH2 rs10744777 polymorphism, which is associated with lower ALDH2 expression, we found 9 of 164 cases versus 3 of 300 controls who carried the BRCA2 p.K3326* variant (OR = 5.66, 95% CI = 1.22-26.2, P = 0.018). This finding supports our hypothesis that the ALDH2-rs10744777 TT genotype may be a significant risk modifier of ESCC in individuals with a BRCA2 p.K3326* mutation

Regulation of albumin by aldosterone.

<p>Mice were administered aldosterone for 0, 24 or 48 hours and paraffin sections from fixed kidneys were subjected to immunohistochemical analysis. A) Representative immunostaining for albumin in control mouse kidneys. B) Albumin immunostaining performed in parallel of a mouse kidney after 24 hours aldosterone administration (Aldo). C) Albumin immunostaining of a mouse kidney after 48 hours aldosterone administration. D) Semiquantified albumin abundance from micrographs similar to A-C after correction for background signal, cell area, and normalization to control values (n = 6). * indicates significant changes relative to control.</p

Renal Type A Intercalated Cells Contain Albumin in Organelles with Aldosterone-Regulated Abundance

<div><p>Albumin has been identified in preparations of renal distal tubules and collecting ducts by mass spectrometry. This study aimed to establish whether albumin was a contaminant in those studies or actually present in the tubular cells, and if so, identify the albumin containing cells and commence exploration of the origin of the intracellular albumin. In addition to the expected proximal tubular albumin immunoreactivity, albumin was localized to mouse renal type-A intercalated cells and cells in the interstitium by three anti-albumin antibodies. Albumin did not colocalize with markers for early endosomes (EEA1), late endosomes/lysosomes (cathepsin D) or recycling endosomes (Rab11). Immuno-gold electron microscopy confirmed the presence of albumin-containing large spherical membrane associated bodies in the basal parts of intercalated cells. Message for albumin was detected in mouse renal cortex as well as in a wide variety of other tissues by RT-PCR, but was absent from isolated connecting tubules and cortical collecting ducts. Wild type I MDCK cells showed robust uptake of fluorescein-albumin from the basolateral side but not from the apical side when grown on permeable support. Only a subset of cells with low peanut agglutinin binding took up albumin. Albumin-aldosterone conjugates were also internalized from the basolateral side by MDCK cells. Aldosterone administration for 24 and 48 hours decreased albumin abundance in connecting tubules and cortical collecting ducts from mouse kidneys. We suggest that albumin is produced within the renal interstitium and taken up from the basolateral side by type-A intercalated cells by clathrin and dynamin independent pathways and speculate that the protein might act as a carrier of less water-soluble substances across the renal interstitium from the capillaries to the tubular cells.</p></div

Extratubular albumin expression.

<p>Immunoperoxidase staining for albumin in the mouse renal cortex with two anti-albumin antibodies. A) Goat anti-albumin labeling of proximal tubules (arrows), as well as peritubular cells (arrow heads). B) Sheep anti-albumin labeling of the same structures. DT = distal tubule, PT = proximal tubule.</p