[The In Vitro Rumen Fermentability on the Processed Vegetable Waste]

This experiment was objected to study the in vitro rumen fermentability on the processed vegetable waste. The study was accomplished by two experiments. The first experiment was aimed to select the best processing for vegetable waste. The vegetable wastes from a traditional market in Semarang City were fermented using Lactobacillus bulgaricus and rumen bolus with cassava waste, rice bran, and maize grain as additives. In each combined treatment of innoculant and additive was fermented in anaerob conditon for 0, 1, 2, and 3 weeks, respectively. In each combined treatment of innoculant, additive, and fermentation time was analyzed for its moisture, crude protein, crude fiber, neutral detergent fiber and acid detergent fiber content. Among the combined treatments were then selected the best processing for vegetable waste ac-cording to the use of L. bulgaricus and rumen bolus, respectively. The selection was determined on the basis of a numerical score for each parameter observed. The results showed that the combination of rice bran of additive and one week of fermentation was the best for fermentation using L. bulgaricus (SSLB). Likewise, the the combination of rice bran of additive and two weeks of fermentation was the best for fermentation using rumen bolus (SSBR). In the second experiment, the processed vegetable wastes from the result of first experiment (SSLB and SSBR) were compared to Pennisetum purpureum (RG), unprocessed vegetable waste from the garbage collecting terminal in Semarang City (SSTPA), and unprocessed vegetable waste from traditional market (SSPT) on the basis of their in vitro rumen fermentability. Parameters of the in vitro rumen fermentability were dry matter (DM) and organic matter (OM) digestibility, rumen production of VFA and N-NH 3 . The test of in vitro rumen fermentability was conducted using cattle rumen liquid and artificial saliva. The results showed that the DM and OM digestibility of RG was lower (P<0,05) than that of SSPT, but there was no siginificant different among SSTPA, SSLB, and RG in their DM and OM digestibility. The in vitro rumen VFA production among RG, SSTPA, SSLB, and SSBR were not different significantly. The in vitro rumen VFA production of SSPT was higher (P<0,05) than that of RG, SSTPA, SSLB, and SSBR. The in vitro rumen NH 3 production among SSPT, SSLB, and SSBR were not different significantly. The in vitro rumen NH 3 production of RG and SSTPA were lower (P<0,05) than that of SSPT, SSLB, and SSBR. The fermen-tation using L.bulgaricus with rice bran for one week was appropriate in processing the vegetable waste. The results could be utilized for further study focusing on the substitution of Pennisetum purpureum with the processed vegetable waste in a ruminant ration. Keywords : processing, vegetable waste, rumen, in vitr

Site-Specific Incorporation of Selenocysteine by Genetic Encoding as a Photocaged Unnatural Amino Acid

Selenocysteine (Sec) is a naturally occurring amino acid that is also referred to as the 21st amino acid. Site-specific incorporation of Sec into proteins is attractive, because the reactivity of a selenol group exceeds that of a thiol group and thus allows site-specific protein modifications. It is incorporated into proteins by an unusual enzymatic mechanism which, in E. coli and other organisms, involves the recognition of a selenocysteine insertion sequence (SECIS) in the mRNA of the target protein. Reengineering of the natural machinery for Sec incorporation at arbitrary sites independent of SECIS elements, however, is challenging. Here we demonstrate an alternative route, whereby a photocaged selenocysteine (PSc) is incorporated as an unnatural amino acid in response to an amber stop codon, using a mutant Methanosarcina mazei pyrrolysyl-tRNA synthetase, Mm PCC2RS, and its cognate tRNACUA. Following decaging by UV irradiation, proteins synthesized with PSc are readily tagged, e.g., with NMR probes to study ligand binding by NMR spectroscopy. The approach provides a facile route for genetically encoded Sec incorporation. It allows the production of pure selenoproteins and the Sec residue enables site-specific covalent protein modification with reagents that would usually react first with naturally occurring cysteine residues. The much greater reactivity of Sec residues allows their selective alkylation in the presence of highly solvent-exposed cysteine residues.Financial support by the Australian Research Council, including a Laureate Fellowship to G.O., is gratefully acknowledged

Trimethylsilyl tag for probing protein-ligand interactions by NMR

Protein-ligand titrations can readily be monitored with a trimethylsilyl (TMS) tag. Owing to the intensity, narrow line shape and unique chemical shift of a TMS group, dissociation constants can be determined from straightforward 1D 1H-NMR spectra not only in the fast but also in the slow exchange limit. The tag is easily attached to cysteine residues and a sensitive reporter of ligand binding also at sites where it does not interfere with ligand binding or catalytic efficiency of the target protein. Its utility is demonstrated for the Zika virus NS2B-NS3 protease and the human prolyl isomerase FK506 binding protein.C.N. and G.O. thank the Alexander von Humboldt Foundation for a Feodor Lynen Fellowship and the Australian Research Council for a Laureate Fellowship, respectively. Financial project support by the Australian Research Council, the Austrian Science Fund (FWF) (DK Molecular Enzymology W901 to K.Z.) and by NAWI Graz is gratefully acknowledged

Pulse EPR-enabled interpretation of scarce pseudocontact shifts induced by lanthanide binding tags

Pseudocontact shifts (PCS) induced by tags loaded with paramagnetic lanthanide ions provide powerful long-range structure information, provided the location of the metal ion relative to the target protein is known. Usually, the metal position is determined by fitting the magnetic susceptibility anisotropy (Δχ) tensor to the 3D structure of the protein in an 8-parameter fit, which requires a large set of PCSs to be reliable. In an alternative approach, we used multiple Gd(3+)-Gd(3+) distances measured by double electron-electron resonance (DEER) experiments to define the metal position, allowing Δχ-tensor determinations from more robust 5-parameter fits that can be performed with a relatively sparse set of PCSs. Using this approach with the 32 kDa E. coli aspartate/glutamate binding protein (DEBP), we demonstrate a structural transition between substrate-bound and substrate-free DEBP, supported by PCSs generated by C3-Tm(3+) and C3-Tb(3+) tags attached to a genetically encoded p-azidophenylalanine residue. The significance of small PCSs was magnified by considering the difference between the chemical shifts measured with Tb(3+) and Tm(3+) rather than involving a diamagnetic reference. The integrative sparse data approach developed in this work makes poorly soluble proteins of limited stability amenable to structural studies in solution, without having to rely on cysteine mutations for tag attachment.Financial support by the Australian Research Council (ARC) and an Australia-Weizmann Making Connections grant is gratefully acknowledged. B. G. thanks the ARC for a Future Fellowship

Synthesis, biological evaluation, and utility of fluorescent ligands targeting the μ-opioid receptor

Fluorescently labeled ligands are useful pharmacological research tools for studying receptor localization, trafficking, and signaling processes via fluorescence imaging. They are also employed in fluorescent binding assays. This study is centered on the design, synthesis, and pharmacological evaluation of fluorescent probes for the opioid receptors, for which relatively few non-peptidic fluorescent probes currently exist. The known μ-opioid receptor (MOR) partial agonist, buprenorphine, was structurally elaborated to include an amidoalkylamine linker moiety that was coupled with a range of fluorophores to afford new fluorescent probes. All compounds proved to be selective MOR antagonists. Confocal fluorescence microscopy studies revealed that the probe incorporating a sulfonated cyanine-5 fluorophore was the most appropriate for imaging studies. This ligand was subsequently employed in an automated fluorescence-based competition binding assay, allowing the pKi values of several well-known opioid ligands to be determined. Thus, this new probe will prove useful in future studies of MOR receptor pharmacology

CEREBROSPINAL FLUID DRAINAGE DEVICES: EXPERIMENTAL CARACTERIZATION

Hydrocephalus is a pathophysiology due to the excess of cerebrospinal fluid in the brain ventricles and it can be caused by congenital defects, brain abnormalities, tumors, inflammations, infections, intracranial hemorrhage and others. Hydrocephalus can be followed by significant rise of intraventricular pressure due to the excess of production of cerebrospinalfluid over the absorption, resulting in a weakening of intellectual functions, serious neurological damage (decreased movement, sensation and functions), critical physical disabilities and even death. A procedure for treatment involves the placement of a ventricular catheter into the cerebral ventricles to divert/drain the cerebrospinal fluid flow to a bag outside of the patient body – provisory treatment known as external ventricular drainage (EVD). Another option is the permanent treatment, internal ventricular drainage (IVD), promoting the cerebrospinal fluid drainage to other body cavity, being more commonly the abdominal cavity. In both cases, EVD and IVD, it is necessary to use of some type of neurological valve in order to control the flow of cerebrospinal fluid. In the present work is proposed an experimental procedure to test the hydrodynamic behavior of a complete drainage system, or parts of them, in order to verify its performance when subjected to pressure gradients found in the human body. Results show that the method is well adapted to quantify the pressure drop in neurological systems

Evaluasi Organoleptik Multinutrien Blok yang dibuat dengan Menggunakan Metode Dingin pada Perbedaan Aras Molases: Organoleptic Evaluation of Multinutrient Block Processed by the Cold Methods on Different Level of Mollases

This research aimed to evaluated and assessed the effect of the molasses level on organoleptic quality of multinutrient block (MnB). The materials used were corn forage, bentonite, urea, salt, molasses, water, blood clamshell and egg shells. The experimental designed used was a Completely Randomized Design with&nbsp; 4 treatments (T0 = MnB Formulation T0 + 0% of molasses, T1 = MnB Formulation T1 + 20% of molasses, T2 = MnB Formulation T2 + 35% of molases, and T3 = MnB Formulation T3 + 50% of molasses) and 4 replications. The variables observed were moisture content, texture, colored and aroma of MnB. The results showed that the addition of molasses had significantly (P &lt;0.05) improved of MnB organoleptic quality. The conclusion for the study, that the multinutrient block formulation with 50% molasses resulted the best organoleptic quality. Key words:&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; molasses, multinutrient block, organoleptic quality DAFTAR PUSTAKA Devendra C &amp; Burns M. 1994. Produksi Kambing di Daerah Tropis. Bogor (ID) : Institut Pertanian Bogor Press Fathia N. 2006. Uji sifat fisik dan mekanik pakan ikan buatan dengan binder tepung tapioka. [skripsi]. Bandar Lampung (ID): Fakultas Pertanian Universitas Lampung. Garcia LO &amp; Restrepo JIR. 1995. Multinutrient Block Handbook (FAO Better Farming Series no. 45). Rome (IT): Food and Agriculture Organization of United Nation. Hermawan, R Sutrisna &amp; Muhtarudin. 2015. Kualitas fisik, kadar air, dan sebaran jamur pada wafer limbah pertanian dengan lama simpan berbeda. Jurnal Ilmu Peternakan Terpadu. 3 (2): 55 – 60 Ismi RS, Pujaningsih RI &amp; Sumarsih S. 2017. Pengaruh penambahan level molases terhadap kualitas fisik dan organoleptik pakan kambing periode penggemukan. Jurnal Ilmu Petenakan. 5 (1): 58-63 Krisnan R &amp; Ginting SP. 2009. Penggunaan Solid Ex-Decanter sebagai binder pembuatan pakan komplit berbentuk pellet: Evaluasi fisik pakan komplit berbentuk pellet. Bogor (ID): Seminar Nasional Teknologi Peternakan dan Veteriner. Bogor Litbang Pertanian Kurnia F, Suhardiman M, Stephani L &amp; Purwadaria T. 2012. Peranan nano-mineral sebagai bahan imbuhan pakan untuk meningkatkan produktivitas dan kualitas produksi ternak. Wartazoa. 22 (4): 187 – 194 Pratama N, Djamas D &amp; Darvina Y. 2016. Pengaruh variasi ukuran partikel terhadap nilai konduktivitas termal papan partikel tongkol jagung. Jurnal Pillar of Physics. 7 (1): 25 – 32 Santi RK, Fatmasari D, Widyawati SD &amp; Suprayogi PS. 2012. Kualitas dan nilai kecernaan in vitro silase batang pisang (Musa paradisiaca) dengan penambahan beberapa akselerator. Jurnal Tropical Animal Husbandry. 1 (1): 15 – 23 Simanihuruk K &amp; Sirait, J. 2010. Silase kulit buah kopi sebagai pakan dasar pada kambing boerka sedang tumbuh. Bogor (ID): Seminar Nasional Teknologi Peternakan dan Veteriner. Litbang Pertanian Syahri M, Retnani Y &amp; Khotijah L. 2018. Evaluasi penambahan binder berbeda terhadap kualitas fisik mineral wafer. Bulletin Makanan Ternak. 16 (1): 24-35 Syukur A &amp; Suharno B. 2014. Bisnis Pembibitan Kambing. Yogyakarta (ID): Penebar Swadaya, Toharmat T, Nurasih E, Nazilah R, Hotimah N, Noerzihad TQ, Sigit NA &amp; Retnani Y. 2005. Sifat fisik pakan kaya serat dan pengaruhnya terhadap konsumsi dan kecernaan nutrien ransum pada kambing. Media Peternakan. 29 (3): 146 – 154 Triyanto E, Prasetiyono BWHE &amp; Mukodiningsih S. 2013. Pengaruh bahan pengemas dan lama simpan terhadap kualitas fisik dan kimia wafer pakan komplit berbasis limbah agroindustri. Jurnal Animal Agriculture. 2 (1): 400 – 409. Warsy, Chadijah S &amp; Rustiah W. 2016. Optimalisasi kalsium karbonat dari cangkang telur untuk produksi pasta komposit. Al-Kimia. 4 (2): 86 - 97 Widiastuti R. 2013. Kualitas pellet berbasis sisa pangan foodcourt dan limbah sayuran fermentasi sebagai bahan pakan fungsional ayam broiler. [tesis] Semarang (ID): Fakultas Peternakan dan Pertanian Universitas Diponegoro Yusmadi, Nahrowi &amp; Ridla M. 2008. Kajian mutu dan palatabilitas silase dan hay ransum komplit berbasis sampah organik primer pada kambing Peranakan Etawah. Agripet. 8 (1): 31 – 38. Zakaria ZA B, Zakaria N &amp; Kasim Z. 2014. Mineral composition of the cockle (Anadara granosa) shells, hard clamp (Meretrix meretrix) shells and corais (Porites spp.): a comparative study. Journal Animal Veterinary Advances. 3 (7): 445 – 447 &nbsp; &nbsp;Penelitian ini bertujuan untuk mengevaluasi dan mengkaji pengaruh aras molases terhadap kualitas organoleptik multinutrien blok (MnB). Bahan yang digunakan dalam penelitian adalah hijauan jagung, bentonit, urea, garam, molases, air, cangkang kerang darah dan cangkang telur. Penelitian dilaksanakan menggunakan rancangan acak lengkap dengan 4 perlakuan aras molases (T0 = Formulasi MnB T0 + 0% molases, T1 = Formulasi MnB T1 + 20% molases, T2 = Formulasi MnB T2 + 35% molases, dan T3 = Formulasi MnB T3 + 50% molasses) dan 4 ulangan. Variabel yang diamati adalah kadar air, tekstur, warna dan aroma MnB. Hasil penelitian menunjukkan bahwa penambahan molases memiliki pengaruh yang signifikan (P&lt;0,05) terhadap peningkatan kualitas organoleptik MnB. Berdasarkan hasil penelitian dapat disimpulkan formulasi multinutrien blok dengan pemberian aras molases sebanyak 50% memiliki kualitas organoleptik paling baik. Kata kunci:&nbsp; aras molasses, kualitas organoleptik, multinutrien blo

Overlapping IgG4 Responses to Self- and Environmental Antigens in Endemic Pemphigus Foliaceus

The etiology of human autoimmune diseases in general remains largely unknown, although the genetic and environmental interplay may be relevant. This applies to the autoimmune diseases of the skin such as the pemphigus phenotypes and others. In this group, there is an endemic form of pemphigus foliaceus [also known as Fogo Selvagem (FS)] where the pathogenic IgG4 autoantibody response to the self-antigen, Desmoglein 1 (Dsg1) cross-react with the LJM11 sand fly salivary gland antigen. In this investigation we dissected the IgG4 autoantibody repertoires utilized by FS patients in response to endogenous self Dsg1 and exogenous LJM11 sand fly antigen. Based on analyses of the genetic clonal signatures of these antibodies, our results indicate that there is a significant overlap between these two responses as all identified IgG4 monoclonal antibodies cross-react to both Dsg1 and LJM11 antigens. Germline H and L chain V gene antibodies generated according to mutated cross-reactive monoclonal antibodies preserved their reactivity to both antigens. Our findings suggest that both Dsg1 autoantigen and LJM11 environmental antigen could be the initial antigenic stimulants for the IgG4 autoimmune responses in FS. These results support our hypothesis that LJM11 antigen plays a substantial role in triggering the IgG4 autoantibody development in FS, and provide new insights on how non-infectious environmental antigen(s) may drive the generation of autoantibodies in IgG4-related autoimmune diseases

Overcoming artificial broadening in Gd³⁺–Gd³⁺ distance distributions arising from dipolar pseudo-secular terms in DEER experiments

By providing accurate distance measurements between spin labels site-specifically attached to bio-macromolecules, double electron–electron resonance (DEER) spectroscopy provides a unique tool to probe the structural and conformational changes in these molecules. Gd3+-tags present an important family of spin-labels for such purposes, as they feature high chemical stability and high sensitivity in high-field DEER measurements. The high sensitivity of the Gd3+ ion is associated with its high spin (S = 7/2) and small zero field splitting (ZFS), resulting in a narrow spectral width of its central transition at high fields. However, under the conditions of short distances and exceptionally small ZFS, the weak coupling approximation, which is essential for straightforward DEER data analysis, becomes invalid and the pseudo-secular terms of the dipolar Hamiltonian can no longer be ignored. This work further explores the effects of pseudo-secular terms on Gd3+–Gd3+ DEER measurements using a specifically designed ruler molecule; a rigid bis-Gd3+-DOTA model compound with an expected Gd3+–Gd3+ distance of 2.35 nm and a very narrow central transition at the W-band (95 GHz). We show that the DEER dipolar modulations are damped under the standard W-band DEER measurement conditions with a frequency separation, Δν, of 100 MHz between the pump and observe pulses. Consequently, the DEER spectrum deviates considerably from the expected Pake pattern. We show that the Pake pattern and the associated dipolar modulations can be restored with the aid of a dual mode cavity by increasing Δν from 100 MHz to 1.09 GHz, allowing for a straightforward measurement of a Gd3+–Gd3+ distance of 2.35 nm. The increase in Δν increases the contribution of the |−5/2〉 → |−3/2〉 and |−7/2〉 → |−5/2〉 transitions to the signal at the expense of the |−3/2 〉 → |−1/2〉 transition, thus minimizing the effect of dipolar pseudo-secular terms and restoring the validity of the weak coupling approximation. We apply this approach to the A93C/N140C mutant of T4 lysozyme labeled with two different Gd3+ tags that have narrow central transitions and show that even for a distance of 4 nm there is still a significant (about two-fold) broadening that is removed by increasing Δν to 636 MHz and 898 MHz.This research was supported by the Israeli Science Foundation (grant 334/14) and made possible in part by the historic generosity of the Harold Perlman Family. D. G. holds the Erich Klieger professorial chair in Chemical Physic

Restauração ecológica em sistemas agroflorestais sucessionais do Vale do Ribeira, São Paulo.

Nas últimas três décadas foi possível observar uma crescente percepção de que a restauração de ecossistemas degradados é necessária, para garantir um mínimo de qualidade de vida para as futuras gerações. O objetivo deste trabalho foi avaliar sistemas agroflorestais sucessionais do Vale do Ribeira, utilizando os indicadores de restauração ecológica da legislação brasileira e os conceitos internacionais estabelecidos pela Sociedade para a Restauração Ecológica, e apontar possíveis soluções para otimização do potencial dos sistemas agroflorestais. Os sistemas agroflorestais sucessionais do Vale do Ribeira demonstram ser práticas eficientes, eficazes e engajadas em restauração ecológica, pois atendem aos indicadores e à maioria dos conceitos empregados como base. Aponta-se a necessidade de construção de políticas públicas voltadas para incentivos normativos e soluções tecnológicas que possibilitem o aumento da geração de renda nestes sistemas. TITLE: Ecological restoration in successional agroforestry systems of Vale do Ribeira, São Paulo State, Brazil ABSTRACT: From the last three decades, there has been a growing perception that the restoration of degraded ecosystems is necessary to guarantee a minimum life quality for future generations. The objective of this work was to evaluate the successional agroforestry systems of Vale do Ribeira, using ecological restoration indicators present in the Brazilian legislation and the in international concepts presented by the Society for Ecological Restoration, and to point out possible solutions to optimize its potential. The successional agroforestry systems in the Vale do Ribeira proved to be efficient, effective and engaged as ecological restoration practices, since they meet the ecological restoration indicators of the legislation and most of the concepts of restoration ecology. Public policies should be developed aiming regulatory incentives and technological solutions to increase income generation by these systems