The Sec1/Munc18 protein Vps45 regulates cellular levels of its SNARE binding partners Tlg2 and Snc2 in Saccharomyces cerevisiae

Intracellular membrane trafficking pathways must be tightly regulated to ensure proper functioning of all eukaryotic cells. Central to membrane trafficking is the formation of specific SNARE (soluble N-ethylmeleimide-sensitive factor attachment protein receptor) complexes between proteins on opposing lipid bilayers. The Sec1/Munc18 (SM) family of proteins play an essential role in SNARE-mediated membrane fusion, and like the SNAREs are conserved through evolution from yeast to humans. The SM protein Vps45 is required for the formation of yeast endosomal SNARE complexes and is thus essential for traffic through the endosomal system. Here we report that, in addition to its role in regulating SNARE complex assembly, Vps45 regulates cellular levels of its SNARE binding partners: the syntaxin Tlg2 and the v-SNARE Snc2: Cells lacking Vps45 have reduced cellular levels of Tlg2 and Snc2; and elevation of Vps45 levels results in concomitant increases in the levels of both Tlg2 and Snc2. As well as regulating traffic through the endosomal system, the Snc v-SNAREs are also required for exocytosis. Unlike most vps mutants, cells lacking Vps45 display multiple growth phenotypes. Here we report that these can be reversed by selectively restoring Snc2 levels in vps45 mutant cells. Our data indicate that as well as functioning as part of the machinery that controls SNARE complex assembly, Vps45 also plays a key role in determining the levels of its cognate SNARE proteins; another key factor in regulation of membrane traffic

Breast adenocarcinoma liver metastases, in contrast to colorectal cancer liver metastases, display a non-angiogenic growth pattern that preserves the stroma and lacks hypoxia

Although angiogenesis is a prerequisite for the growth of most human solid tumours, alternative mechanisms of vascularisation can be adopted. We have previously described a non-angiogenic growth pattern in liver metastases of colorectal adenocarcinomas (CRC) in which tumour cells replace hepatocytes at the tumour-liver interface, preserving the liver architecture and co-opting the sinusoidal blood vessels. The aim of this study was to determine whether this replacement pattern occurs during liver metastasis of breast adenocarcinomas (BC) and whether the lack of an angiogenic switch in such metastases is due to the absence of hypoxia and subsequent vascular fibrinogen leakage. The growth pattern of 45 BC liver metastases and 28 CRC liver metastases (73 consecutive patients) was assessed on haematoxylin- and eosin-stained tissue sections. The majority of the BC liver metastases had a replacement growth pattern (96%), in contrast to only 32% of the CRC metastases (P<0.0001). The median carbonic anhydrase 9 (CA9) expression (M75 antibody), as a marker of hypoxia, (intensity x % of stained tumour cells) was 0 in the BC metastases and 53 in the CRC metastases (P<0.0001). There was CA9 expression at the tumour-liver interface in only 16% of the BC liver metastases vs 54% of the CRC metastases (P=0.002). There was fibrin (T2G1 antibody) at the tumour-liver interface in only 21% of the BC metastases vs 56% of the CRC metastases (P=0.04). The median macrophage count (Chalkley morphometry; KP-1 anti-CD68 antibody) at the interface was 4.3 and 7.5, respectively (P<0.0001). Carbonic anhydrase 9 score and macrophage count were positively correlated (r=0.42; P=0.002) in all metastases. Glandular differentiation was less in the BC liver metastases: 80% had less than 10% gland formation vs only 7% of the CRC metastases (P<0.0001). The liver is a densely vascularised organ and can host metastases that exploit this environment by replacing the hepatocytes and co-opting the vasculature. Our findings confirm that a non-angiogenic pattern of liver metastasis indeed occurs in BC, that this pattern of replacement growth is even more prevalent than in CRC, and that the process induces neither hypoxia nor vascular leakage

PERUBAHAN KEKERASAN MIKRO DENTIN SETELAH APLIKASI EDTA 15%, EDTAC 15% DAN KOMBINASI EDTA 15%- UREA PEROKSIDA PADA TIGA SEGMEN AKAR GIGI

The successful of endodontic treatment affected by an effective disinfectant. Chelating agent such as ethylenediaminetetraacetic acid (EDTA), EDTAC and EDTA combined with urea peroxide were an effective disinfectant to remove smear layer.The propose of this study was to evaluate the effect of chelating agents 15% EDTA, 15% EDTAC and 15% EDTA combined with urea peroxide on the microhardness of coronal 1/3, middle 1/3 and apical 1/3 segments of root canal dentin. Thirty single rooted premolars were sectioned horizontally at cervical level. After root canal treated the roots were sectioned into three segments which then embedded in acrylic resin. Ninety sectioned root segments were devided into 3 groups of 30 Each group was treated with 15% EDTA, 15% EDTAC and 15% EDTA combined with urea peroxide respectively for 5 minutes. The groups were then devided into 3 subgroups which were coronal 1/3 segments, middle 1/3 segments and apical 1/3 segments. Root canal dentin microhardness measurement was carried out using Vickers Microhardness Tester before and after application of chelating agent. The data were statistically analized using Kruskall Wallis and Mann-Whitney U test at 95% confidence level. The results showed that there were reduction on root canal dentin microhardness after treated wi

Analisis Usaha Peternakan Sapi Perah Di Kelompok Kud Semen Pada Masa Pandemi Covid-19 Di Desa Semen Kecamatan Gandusari Kabupaten Blitar

Tujuan penelitian ini adalah untuk mengetahui biaya produksi, penerimaan, keuntungan, dan kelayakan usaha berdasarkan nilai Break Even Point (BEP) dan Return Per Cost (R/C) pada usaha peternakan sapi perah di Desa Semen Kecamatan Gandusari Kabupaten Blitar. Kabupaten, Provinsi Jawa Timur berdasarkan skala kepemilikan ternak. Metode yang digunakan dalam penelitian ini adalah metode survey dengan menggunakan wawancara atau observasi tidak langsung melalui telepon atau alat komunikasi lainnya. Wawancara dilakukan untuk mengumpulkan data yang dibutuhkan kepada pihak terkait sesuai dengan judul penelitian yang dilakukan. Biaya produksi, pendapatan, dan keuntungan berturut-turut dalam skala kecil Rp1.536.100/AU/bulan, skala menengah Rp1.652.666/AU/bulan, skala menengah Rp1.652.666/AU /bulan, 1.666.557 IDR/AU/bulan, 1.679.812 IDR/AU/bulan, skala besar 116.566 IDR/AU/bulan, 168.990 IDR/AU/bulan, 259.885 IDR/AU/bulan. BEP Produk, Harga BEP, dan R/C Ratio masing-masing pada skala kecil 269 liter/AU/bulan, Rp 5.078/liter, 1,07, skala menengah 262 liter/AU/bulan, Rp 4.963/liter, 1, 11, skala besar 249 liter/AU/bulan, Rp3.424/liter, 1,18. Kesimpulan dari penelitian ini adalah bahwa semakin besar bisnisnya, semakin besar keuntungannya. Hal ini sesuai dengan skala peternakan skala III yang memiliki keuntungan terbesar dibanding skala I dan II

Uji Daya Hambat Ekstrak Kasar Daun Keji Beling (Strobilanthes crispus) terhadap Bakteri Edwardsiella tarda secara In Vitro

Peningkatan produksi perikanan dapat dilakukan dengan intensifikasi budidaya, namun usaha ini tidak terlepas dari ancaman serangan penyakit pada ikan. Salah satu penyakit yang banyak ditemukan adalah Edwardsiellosis yang disebabkan karena infeksi bakteri Edwardsiella tarda. Umumnya digunakan antibiotik sebagai pengobatan, namun penggunaan antibiotik dapat memicu dampak negatif seperti menumpuknya residu antibiotik, resistensi bakteri, pencemaran lingkungan dan lainnya. Upaya alternatif yang dapat dilakukan adalah dengan menggunakan bahan alami yang mengandung senyawa antibakteri seperti daun keji beling (S. crispus). Penelitian ini bertujuan untuk mengetahui daya hambat ekstrak kasar daun keji beling (S. crispus) terhadap bakteri E. tarda secara in vitro. Penelitian dilakukan di Laboratorium Sentral Ilmu Hayati (LSIH), Universitas Brawijaya, Malang pada bulan Desember 2021 – Januari 2022. Metode penelitian yang digunakan adalah metode eksperimental untuk mengetahui secara langsung pengaruh pemberian berbagai dosis ekstrak kasar daun keji beling (S. crispus) terhadap daya hambat bakteri E. tarda yang ditandai dengan luasan zona bening yang terbentuk. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan 5 perlakuan dan 3 ulangan. Dosis perlakuan ekstrak kasar daun keji beling (S. crispus) yaitu: A (30 ppm), B (60 ppm), C (90 ppm), D (120 ppm) dan E (150 ppm). Hasil penelitian menunjukkan bahwa ekstrak kasar daun keji beling (S. crispus) memberikan pengaruh beda nyata terhadap pertumbuhan bakteri E. tarda. Pengamatan zona hambat dilakukan setelah inkubasi selama 24 dan 48 jam. Hasil pengamatan setelah 24 jam menunjukkan bahwa dosis ekstrak dengan rata-rata diameter zona hambat tertinggi adalah pada perlakuan E (150 ppm) sebesar 7,701 ± 0,26 mm dan dosis ekstrak dengan rata-rata zona hambat terendah pada perlakuan A (30 ppm) sebesar 6,207 ± 0,30 mm. Pengamatan setelah inkubasi 48 jam menunjukkan bahwa zona hambat membesar yang menandakan sifat antibakteri sebagai bakterisidal. Penelitian ini menunjukkan bahwa hubungan antara pemberian ekstrak kasar daun keji beling (S. crispus) terhadap pertumbuhan bakteri E. tarda membentuk pola linier dengan persamaan y = 0,0126x + 5,979 dan koefisien nilai determinasi (R2) sebesar 0,6268. Kesimpulan yang didapat adalah pemberian ekstrak kasar daun keji beling (S. crispus) efektif sebagai bahan antibakteri alami bakteri E. tarda dengan dosis terbaik adalah pada perlakuan E (150 ppm). Bahan alami ini bersifat bakterisidal, yaitu mampu membunuh pertumbuhan bakteri. Semakin tinggi dosis yang diberikan, maka semakin besar zona hambat yang dihasilkan