SELECTION OF LOCATORS IN AUTOMATED DESIGN OF FIXTURES FOR LOCATING WORKPIECES DURING MACHINING

Using the systematic approach, the possible basing schemes of the workpieces in the machining attachments have been discovered and systematized with a view to their use in automated design. The analysis shows that the use of the proposed systematization in computer-aided design is significantly more rational, since the structure reflects the sequence of choice of the optimal basing scheme - analysis of the theoretical basing scheme, analysis of the geometry of the workpiece, formation of the possible list of the basing schemes. An analysis was performed with the help of which the criteria for geometric compatibility were revealed, allowing to make a choice of a basing scheme, satisfying the geometric shape of the workpiece. An algorithm for selection of locators has been developed. The development is part of a system for automated design of fixtures for locating of workpieces during machining.

Impact of multi-metals (Cd, Pb and Zn) exposure on the physiology of the yeast Pichia kudriavzevii

Metal contamination of the environment is frequently associated to the presence of two or more metals. This work aimed to study the impact of a mixture of metals (Cd, Pb and Zn) on the physiology of the non-conventional yeast Pichia kudriavzevii. The incubation of yeast cells with 5 mg/l Cd, 10 mg/l Pb and 5 mg/l Zn, for 6 h, induced a loss of metabolic activity (assessed by FUN-1 staining) and proliferation capacity (evaluated by a clonogenic assay), with a small loss of membrane integrity (measured by trypan blue exclusion assay). The staining of yeast cells with calcofluor white revealed that no modification of chitin deposition pattern occurred during the exposure to metal mixture. Extending for 24 h, the exposure of yeast cells to metal mixture provoked a loss of membrane integrity, which was accompanied by the leakage of intracellular components. A marked loss of the metabolic activity and the loss of proliferation capacity were also observed. The analysis of the impact of a single metal has shown that, under the conditions studied, Pb was the metal responsible for the toxic effect observed in the metal mixture. Intracellular accumulation of Pb seems to be correlated with the metals toxic effects observed.The authors thank the FCT Strategic Project PEst-OE/EQB/LA0023/2013 and the Project "BioInd-Biotechnology and Bioengineering for improved Industrial and Agro-Food processes" (NORTE-07-0124-FEDER-000028), Co-funded by the Programa Operacional Regional do Norte (ON.2-O Novo Norte), QREN, FEDER. Manuela D. Machado gratefully acknowledges the post-doctoral grant from FCT (SFRH/BPD/72816/2010). Vanessa A. Mesquita gratefully acknowledges the grant from Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES). The authors also thank to Doctor Rosane Freitas Schwan to offer the yeast strain and to Doctor Helena M.V.M. Soares, from the Faculty of Engineering of Porto University, for the use of analytical facilities (AAS with flame atomization and AAS with electrothermal atomization)

LAGU KIDDUNG DALEM DALAM UPACARA ADAT NYONGKOLAN SUKU SASAK DI KABUPATEN LOMBOK TENGAH (BENTUK PENYAJIAN DAN BENTUK LAGU)

ABSTRAKLagu Kiddung Dalem merupakan satu lagu tradisi dalam musik tradisional Gendang Beleq, yang disajikan secara instrumental di rumah pengantin perempun, dalam prosesi upacara adat Nyongkolan suku Sasak pulau Lombok, Nusa Tenggara Barat. Lagu Kiddung Dalem digunakan sebagai bentuk ekspresi dari rasa syukur, kegembiraan, kekeluargaan dan juga untuk menyampaikan petuah–petuah adat dalam suku Sasak. Bahkan digunakan sebagai sarana melegitimasi kelangsungan kehidupan tradisi adat melalui prosesi adat pernikahan. Sehingga akan sangat menarik dilakukan penelitian dengan rumusan masalah (1) Bagaimanakah bentuk penyajian lagu Kiddung Dalem dalam prosesi upacara adat Nyongkolan di suku Sasak, dan (2) Bagaimanakah bentuk lagu Kiddung Dalem dalam upacara adat Nyongkolan suku Sasak.Kajian pustaka yang digunakan didalam penelitian, mengacu sumber buku–buku yang berkaitan mengenai bentuk penyajian seni pertunjukan dan bentuk sertaa struktur didalam lagu. Penelitian yang digunakan, menggunakan metode penelitian kualitatif, dengan lokasi penelitian di kelurahan Gerunung, kabupaten Lombok Tengah, Nusa Tenggara Barat. Hasil penelitian berupa sajian data bersifat diskriptif tentang bentuk penyajian dan bentuk lagu Kiddung Dalem yang disajikan saat upacara adat Nyongkolan berlangsung. Dalam penelitian tentang bentuk penyajian, peneliti menemukan data meliputi, (1) Instrumentasi, yaitu ragam jenis alat musik beserta pola–pola sajain yang dimainkan oleh 17 pemusik atu Sekhe. (2) Setting pertunjukan, dilakukan di halaman rumah pengantin perempuan ketika prosesi adat Nyongkolan berlangsung. Sehingga panggung pertunjukan, berbentuk panggung terbuka dan, (3) Tata busana yang digunakan adalah baju Godeq Nongkeq yang merupakan pakaian adat laki–laki suku Sasak.Penelitian tentang bentuk lagu Kiddung Dalem meliputi penemuan data tentang pembagian dua bagian lagu yang terdiri dari 58 birama dan terbagi dalam 8 frase tanya dan 8 jawab serta memiliki 32 motif. Pada bagian pertama disebut dengan Kabor yang terdiri dari 4 bagian dan bagian kedua yaitu Pelayon sebagai bagian lagu utama. Kata Kunci: Kiddung Dalem, Bentuk Penyajian, Bentuk Lagu

Produksi VCO Secara Tradisonal sebagai Obat Alami dalam Meningkatkan Pendapatan Masyarakat Desa Watukamba

Virgin Coconut Oil is an oil that is processed from the extraction carried out from fresh coconut fruit which is processed in various ways, one of which is by using traditional methods, namely the natural processing process, namely by photosynthesis. The purpose of making Virgin Coconut Oil is to take advantage of the potential of nature in agriculture in increasing the empowerment of the creative economy of coastal communities during the Covid-19 pandemic. The method used is socialization and training on the process of making Virgin Coconut Oil (VCO). The result obtained is that the people of Watukamba Village realize the importance of utilizing the natural potential in agriculture and being able to produce VCO independently to increase the economic income of the community. The conclusion after community service activities during the pandemic is still felt the benefits by the community and the sustainability of virgin coconut oil (VCO) processing

MATHIMATICAL MODEL TO PREDICT THE DESIGN ANI) PERFORMANCE OF HIGH.SPEED TURBOPUMPS

In this work a high-speed turbopumps of (22000 rpm) rotational speed, which are oxidizer and fuel turbopumps, are designed. The design requirement for mass flow rate and outlet pressure are (12.5 kg/s) and (113×10 Pa) respectively for the oxidizer turbopump, and (3.5 kg/s) and (95x10 Pa) respectively for the fuel turbopump. The design work of centrifugal pump is divided into impeller and volute design works. The impeller design work is performed by the streamline curvature and quasi-orthogonal technique based on hub-to-shroud mean stream surface between two blades with assistance of experimental design coefficients and relations for first prediction. The volute is designed by using the method of constant angular momentum, where in this method the tangential and radial velocities are considered uniform around the impeller. The numerical results are compared with experimental data of the actual pumps, and it is found that these results agree well with the real designs. The output of the design program is used as an input to the performance program. The performance prediction is achieved by three dimensional, steady, incompressible and viscous flow analyses through the blade passage. The ANSYS program has been employed to solve these equation in order to predict the flow characteristics at design and off-design conditions. The analysis is done at many operation conditions, which have the range of flow coefficient ratio (ФФ) from (0.4) to (1.8), and the results are compared with experimental results for previous work to make sure the validity of the analysis program at design and off-design conditions

Copper-Dependent Trafficking of the Ctr4-Ctr5 Copper Transporting Complex

In Schizosaccharomyces pombe, copper uptake is carried out by a heteromeric complex formed by the Ctr4 and Ctr5 proteins. Copper-induced differential subcellular localization may play a critical role with respect to fine tuning the number of Ctr4 and Ctr5 molecules at the cell surface.We have developed a bimolecular fluorescence complementation (BiFC) assay to analyze protein-protein interactions in vivo in S. pombe. The assay is based on the observation that N- and C-terminal subfragments of the Venus fluorescent protein can reconstitute a functional fluorophore only when they are brought into tight contact. Wild-type copies of the ctr4(+) and ctr5(+) genes were inserted downstream of and in-frame with the nonfluorescent C-terminal (VC) and N-terminal (VN) coding fragments of Venus, respectively. Co-expression of Ctr4-VC and Ctr5-VN fusion proteins allowed their detection at the plasma membrane of copper-limited cells. Similarly, cells co-expressing Ctr4-VN and Ctr4-VC in the presence of Ctr5-Myc(12) displayed a fluorescence signal at the plasma membrane. In contrast, Ctr5-VN and Ctr5-VC co-expressed in the presence of Ctr4-Flag(2) failed to be visualized at the plasma membrane, suggesting a requirement for a combination of two Ctr4 molecules with one Ctr5 molecule. We found that plasma membrane-located Ctr4-VC-Ctr5-VN fluorescent complexes were internalized when the cells were exposed to high levels of copper. The copper-induced internalization of Ctr4-VC-Ctr5-VN complexes was not dependent on de novo protein synthesis. When cells were transferred back from high to low copper levels, there was reappearance of the BiFC fluorescent signal at the plasma membrane.These findings reveal a copper-dependent internalization and recycling of the heteromeric Ctr4-Ctr5 complex as a function of copper availability

Transcriptome response to heavy metal stress in Drosophila reveals a new zinc transporter that confers resistance to zinc

All organisms are confronted with external variations in trace element abundance. To elucidate the mechanisms that maintain metal homeostasis and protect against heavy metal stress, we have determined the transcriptome responses in Drosophila to sublethal doses of cadmium, zinc, copper, as well as to copper depletion. Furthermore, we analyzed the transcriptome of a metal-responsive transcription factor (MTF-1) null mutant. The gene family encoding metallothioneins, and the ABC transporter CG10505 that encodes a homolog of ‘yeast cadmium factor’ were induced by all three metals. Zinc and cadmium responses have similar features: genes upregulated by both metals include those for glutathione S-transferases GstD2 and GstD5, and for zinc transporter-like proteins designated ZnT35C and ZnT63C. Several of the metal-induced genes that emerged in our study are regulated by the transcription factor MTF-1. mRNA studies in MTF-1 overexpressing or null mutant flies and in silico search for metal response elements (binding sites for MTF-1) confirmed novel MTF-1 regulated genes such as ferritins, the ABC transporter CG10505 and the zinc transporter ZnT35C. The latter was analyzed in most detail; biochemical and genetic approaches, including targeted mutation, indicate that ZnT35C is involved in cellular and organismal zinc efflux and plays a major role in zinc detoxification

Zinc-Regulated DNA Binding of the Yeast Zap1 Zinc-Responsive Activator

The Zap1 transcription factor of Saccharomyces cerevisiae plays a central role in zinc homeostasis by controlling the expression of genes involved in zinc metabolism. Zap1 is active in zinc-limited cells and repressed in replete cells. At the transcriptional level, Zap1 controls its own expression via positive autoregulation. In addition, Zap1's two activation domains are regulated independently of each other by zinc binding directly to those regions and repressing activation function. In this report, we show that Zap1 DNA binding is also inhibited by zinc. DMS footprinting showed that Zap1 target gene promoter occupancy is regulated with or without transcriptional autoregulation. These results were confirmed using chromatin immunoprecipitation. Zinc regulation of DNA binding activity mapped to the DNA binding domain indicating other parts of Zap1 are unnecessary for this control. Overexpression of Zap1 overrode DNA binding regulation and resulted in constitutive promoter occupancy. Under these conditions of constitutive binding, both the zinc dose response of Zap1 activity and cellular zinc accumulation were altered suggesting the importance of DNA binding control to zinc homeostasis. Thus, our results indicated that zinc regulates Zap1 activity post-translationally via three independent mechanisms, all of which contribute to the overall zinc responsiveness of Zap1

A Modified Protocol for Bisulfite Genomic Sequencing of Difficult Samples

The bisulfite genomic sequencing protocol is a widely used method for analyzing DNA methylation. It relies on the deamination of unmethylated cytosine residues to uracil; however, its high rates of DNA degradation and incomplete cytosine to uracil conversion often lead to failed experiments, uninformative results, and false positives. Here, we report the addition of a single-step multiple restriction enzyme digestion (MRED) designed to differentially digest polymerase chain reaction products amplified from unconverted DNA while leaving those of converted DNA intact. We show that for our model system, RARB2 P2 promoter, use of MRED increased informative sequencings ninefold, and MRED did not alter the clonal representation in one fully methylated cell line, H-596, treated or not with 5-azadeoxycytidine, a methylation inhibitor. We believe that this method may easily be adapted for analyzing other genes and provide guidelines for selecting the most appropriate MRED restriction enzymes

Regulation of Alr1 Mg Transporter Activity by Intracellular Magnesium

Mg homeostasis is critical to eukaryotic cells, but the contribution of Mg transporter activity to homeostasis is not fully understood. In yeast, Mg uptake is primarily mediated by the Alr1 transporter, which also allows low affinity uptake of other divalent cations such as Ni2+, Mn2+, Zn2+ and Co2+. Using Ni2+ uptake to assay Alr1 activity, we observed approximately nine-fold more activity under Mg-deficient conditions. The mnr2 mutation, which is thought to block release of vacuolar Mg stores, was associated with increased Alr1 activity, suggesting Alr1 was regulated by intracellular Mg supply. Consistent with a previous report of the regulation of Alr1 expression by Mg supply, Mg deficiency and the mnr2 mutation both increased the accumulation of a carboxy-terminal epitope-tagged version of the Alr1 protein (Alr1-HA). However, Mg supply had little effect on ALR1 promoter activity or mRNA levels. In addition, while Mg deficiency caused a seven-fold increase in Alr1-HA accumulation, the N-terminally tagged and untagged Alr1 proteins increased less than two-fold. These observations argue that the Mg-dependent accumulation of the C-terminal epitope-tagged protein was primarily an artifact of its modification. Plasma membrane localization of YFP-tagged Alr1 was also unaffected by Mg supply, indicating that a change in Alr1 location did not explain the increased activity we observed. We conclude that variation in Alr1 protein accumulation or location does not make a substantial contribution to its regulation by Mg supply, suggesting Alr1 activity is directly regulated via as yet unknown mechanisms