Special Reagents Research Articles

A success story in a low-resource setting: Advancing cancer diagnostics in underserved region

Abstract Introduction/Objective Shefaa Al Orman Hospital (SOH) is the first oncology hospital in Luxor. Since its inception in 2016, the hospital has served approximately 45,000 patients from Luxor and other governorates. The hospital’s lab provides excellent routine laboratory services. However, there is a pressing demand for molecular genetics and cytogenetics, that is necessary for diagnosis of hematological malignancies. This strategic expansion faced several challenges: lack of expertise and the need for special expensive equipment and reagents. Methods/Case Report The project started with collaboration with oncologists to convene on a simplified panel with the aim of prioritizing clinically relevant tests. At first, these investigations were referred to a specialized oncology hospital in Cairo. This entailed 2 problems: transport of samples led to deterioration of the quality of some samples and delay in results reporting. In the second phase, experienced consultants from Cairo were enlisted to provide training and mentorship to local staff. Infrastructure setup included the allocation of dedicated laboratory space and the procurement of specialized equipment. Test method selection was used to achieve successful transition. In cytogenetics, we started with FISH technique which, while more expensive, gave rapid and reliable results. After gaining experience, the technique of conventional karyotyping was established. On the other hand, in molecular genetics, we resorted to custom made reagent mix preparation, instead of commercial kits. This approach, while more laborious, is less expensive. Results (if a Case Study enter NA) With the gradual successful in-house setup of these tests, there has been a notable reduction of out-sourced samples to 5%. This has led to shorter result turnaround times. Cytogenetics and FISH improved from 4 to 1 week, while molecular genetics decreased from 4 to 2 weeks. Moreover, the local processing of samples has minimized the risk of sample damage during transportation, ensuring the integrity of test results. Conclusion The establishment of the Molecular Genetics and Cytogenetics Laboratory at SOH represents a significant milestone. The hospital has successfully overcome logistical challenges and enhanced diagnostic capabilities for hematological malignancies, contributing to better patient outcomes and satisfaction. Moving forward, the continued investment in infrastructure, personnel training, and quality assurance measures will further strengthen the hospital’s capacity to provide high-quality cancer care to the community.

B-037 Zeon original coating technology COP microplates for cell imaging

Abstract Background Zeon has developed high quality microplate made of Cyclo Olefin Polymer (COP) and special coat reagent for cell culture for Aurora microplates. The new 96 well-microplates have best-in-class properties for analysis of cultured cells can reduce the background of fluorescence images absolutely and excellent bottom flatness for observation of target. This enables to obtain high-precision data for cell analysis. Also, we developed Zeon original coating (ZOC) reagent specialized for COP surface. It is suitable for various cells including neurons derived from human induced pluripotent stem cells (iPSC), primary cells, general cell lines, and so on. ZOC coated COP microplates (ZOC plates) are fully-precoated and ready-to-use. Methods We evaluated the following points regarding ZOC plates. The cells used were derived from human iPSC. [1. Storage stability] ZOC plates were stored at room temperature for a long time. After that, cardiomyocyte cells were cultured for 3 days on the plates. The cell adhesion and beat rate were evaluated. [2. Neurite extension] Dopaminergic neurons were cultured for 14 days, then the neurite extension was evaluated. For comparison, a conventional poly-D-lysine* (PDL) coating was used. [3. Lot-to-lot variation] COP microplates coated with different lots of ZOC reagents were prepared. Glutamatergic neurons were cultured for 14 days on those plates, then the neurite extension and cell morphology were evaluated. *The typical coating reagent applied for neuronal culture. Results [1.] ZOC plates stored over 12 months at room temperature showed excellent cell adhesion, and the beat rate was equivalent to prepared just before use. [2.] ZOC plates tended to extend neurites more than PDL coated plates. [3.] No variation between lots was observed on ZOC plates regarding the neurite extension and cell morphology. Conclusions We suggest applying the combination of high accuracy for cell imaging using COP microplates and high performance for cell culture using ZOC can provide high precision data at bioassay research. This would be a significant value for development of drug screening processes at pharmaceutical company.

Synthesis of β-Silyl Amines via Merging Photoinduced Energy and Hydrogen Atom Transfer in Flow.

The development of efficient methods for synthesizing β-silyl amines has long been a significant goal in organic synthesis. Previous methods mainly relied on the use of prefunctionalized substrates or special reagents. Herein, we present a visible-light-promoted synthesis approach for β-silyl amines, utilizing a combination of photoinduced energy and hydrogen atom transfer processes. Using flow chemistry technology, a variety of valuable skeletons, including β-silyl amines and α-amino esters, can be produced from readily available feedstocks such as hydrosilanes and simple alkanes. Moreover, the strategy's full-process fluidized production capability highlights its potential for industrial-scale manufacturing. Mechanistic studies revealed that oxime esters can act as radical precursors as well as hydrogen atom transfer reagents.

STUDYING INDICATORS OF CIRCADIAN RHYTHMS OF MACROELEMENTS CONCENTRATION IN HUMAN SALIVA UNDER NORMAL CONDITIONS

Seasonal and circadian research in the field of chronobiology and chronomedicine is currently of particular interest to scientists. Chronobiology has discovered previously unknown approaches to the diagnosis, treatment and prevention of diseases. For biological objects, as an open thermodynamic system, in their development have absorbed all the laws of the rhythmically oscillating external world and interact with it with the help of these vibrations. Therefore, at the present stage of the development of chronobiology, any research that fills, like bricks, still empty niches of new knowledge reflects the advanced frontiers of natural science and is included in the creation of a future general theory of biology. This is confirmed by a number of discoveries of scientists in the field of physics, physiology and medicine. Thus, a group of scientists became laureates of the Nobel Prize in physics in 2017 for the discovery of gravitational waves, and in the field of physiology and medicine, a number of scientists (Hall, Rosbash, Young) became Nobel laureates for a series of works on the topic: "Studying the mechanisms of the circadian dynamics of the body." Today, chronobiology can characterize periodic fluctuations of, perhaps, all organs, all body systems. A large amount of data has been accumulated on the daily and seasonal rhythms of the digestive and urinary tract organs, respiratory and cardiovascular systems, brain, etc. A violation of biorhythms can lead to the development of defects in oral and maxillofacial tissues, the development of inflammation and tumors, which affects cell proliferation, apoptosis, oxidative stress, etc. The main purpose of this work was to study the characteristics of circadian rhythms of macroelement concentrations in human saliva under normal conditions. A feature of the research work is the determination of the concentration of macroelements in saliva in the summer. The saliva of 12 healthy volunteers was taken as the study material. Age 22-30 years. The method of spectrophotometry was used in this research work. The research work was carried out in the summer at the Department of Biophysics, Biomedicine and Neuroscience of the al-Farabi Kazakh National University and at the City Clinical Hospital №1 in Almaty. During the study, a saliva sample was collected on 1.5 ml of Eppendorf. All samples were stored in the freezer. Before the experiment, the material was soaked at room temperature for 30-40 minutes. Using the spectrophotometric method, we determined the concentration of macroelements in saliva using a set of special reagents. We have identified the features of the daily dynamics of the concentration of macroelements in the saliva of healthy people in summer: K, Na, Ca, Mg and P. The concentration of macroelements varied widely throughout the day. As a result of the study, the concentration index of K during the day normally ranges from 528±36 µg/ml to 634±42 µg/ml; Na - from 153.4±19.8 µg/ml to 144.9±25.1 µg/ml; Ca-from 35.4±4.0 µg/ml to 45.5±5.1 µg/ml; Mg-from 2.12±0.15 µg/ml to 4.02±0.25 µg/ml and P-from 134.5±7.7 µg/ml to 151.6±10.1 µg/ml. Summing up, we have identified the features of the daily dynamics of the concentration of macroelements in the saliva of healthy people in summer. In summer, the daily dynamics of the concentration of macroelements in saliva ranged from 2.12±0.15 µg/ml (Mg) to 634±12 µg/ml (K).

Practical preparation of unsaturated very-long-chain fatty acids (VLCFAs) and very-long-chain alkene pollinator attractants

To prepare very-long-chain fatty acids and alkenes (VLCFAs and VLC alkenes) that are known pollinator attractants for sexually deceptive orchids, and biosynthetic precursors thereof, we applied a methodology allowing us to prepare monounsaturated VLCFAs with chain lengths up to 28 carbons and VLC alkenes up to 31 carbons. We implemented a coupling reaction between commercially available terminal alkynes and bromoalkanoic acids to prepare VLCFAs, allowing the products to be formed in two steps. For VLC alkenes, with many alkyltriphenylphosphonium bromides commercially available, we applied a Wittig reaction approach to prepare (Z)-configured monoenes in a single step. Using practical methods not requiring special reagents or equipment, we obtained 11 VLCFAs in > 90% isomeric purity, and 17 VLC alkenes in > 97% isomeric purity. Such general and accessible synthetic methods are essential for chemical ecology and biochemistry research to aid researchers in unambiguously identifying isolated semiochemicals and their precursors.

Wheat as a test object for determining the degree of soil contamination with petroleum products

Abstract Currently, the oil and gas industry remains the leader in meeting the global need for energy resources. The high intensity of hydrocarbon production naturally leads to an increase in the volume of petrochemical waste that ends up in the natural environment. In this regard, the implementation of toxicological assessment of oil-containing soils and water bodies is one of the most important environmental tasks. Biotesting allows for a quick integral assessment of the properties of a contaminated environment. This method of toxicity analysis is economical and does not require the use of special chemical reagents and equipment. The seeds of higher vegetation are the most accessible, easy to use and universal test objects. However, it is necessary to use only species that are highly sensitive to pollutants. This article is based on the results of a study of the phytotoxicity of gray forest soil contaminated with petroleum products using wheat. Experiments have shown that soil contaminated with oil waste has a strong inhibitory effect on wheat seedlings. It was found that small doses of petroleum products (0.31% oil) have a stimulating effect on seed germination in the analyzed soil.

Analysis of test results of the P-20 multi-rotor UAV when spraying pesticides in precision agriculture

Abstract The article provides an overview of the current results of using multi-rotor UAVs when spraying pesticides in precision agriculture. It is noted that, as part of improving the technology for using UAVs, it is necessary to continue work on optimizing and testing the parameters of UAV transport and technological cycles together with a reasonable choice of special reagents and their combinations to improve the spraying effect and increase the level of use of pesticides. This will directly affect the improvement of the quality and efficiency of precision farming systems. The analysis presented in the work is based on testing the parameters of the transport and technological cycle of a multi-rotor UAV of the P-20 type, which was used to spray pesticides on cotton fields to combat aphids and spider mites on cotton at the flowering and boll setting stages. The results are considered that make it possible to analyze and compare the laws of uniform and penetrating deposition and drift of droplets in cotton canopies at different UAV flight altitudes. It has been shown that spraying small volumes of agrochemicals using UAVs at low altitudes differs from operations performed using modern manned aircraft or boom sprayers. The flight altitude of the UAV has a significant impact on the deposition and drift of droplets. The results of studies related to elucidating the effect of defoliant adsorption on cotton leaves are also noted. The adsorption of acetamiprid and spirodiclofen was studied. Methods for statistical processing of the obtained experimental data are presented, in particular, to test the significance of the influence of flight altitude on the experimental results.

Analysis of the application of flow-deflecting technologies in order to increase oil recovery

This article discusses flow-deflecting technologies as a method of increasing oil recovery based on the injection of limited volumes of special reagents into injection wells. This technology allows you to: reduce the permeability of highly permeable layers of the formation; align the well intake along the formation section; create a more uniform displacement front; reduce water breakthroughs to the producing well. Flow-deflecting technologies are used at the final stage of field development or in solving problems related to repair and insulation work. Also in this article, an analysis of the effectiveness of flow-deflecting technologies using various polymers is carried out. The results of experimental and field studies of three new technologies based on the use of a chemical agent, for example, POLY-T-101, hydrophobic emulsions and gel-acid compositions are presented during work on limiting water flows and redistributing the directions of filtration flows in terrigenous carbonate reservoirs at reservoir temperatures from 20 to 1000 C. The issues of the mechanism of action of chemical reagents on productive deposits and prospects for the application of the developed technologies at problematic oil field facilities are considered. The results of field tests are presented.

Quantitative 1H NMR Spectroscopy Method for Determination of Anthraquinone Derivatives in Extracts from Rubia tinctorum L. Roots and Rhizomes

Quantitative 1H NMR Spectroscopy Method for Determination of Anthraquinone Derivatives in Extracts from Rubia tinctorum L. Roots and Rhizomes

A slow but steady nanoLuc: R162A mutation results in a decreased, but stable, nanoLuc activity

NanoLuc (NLuc) luciferase has found extensive application in designing a range of biological assays, including gene expression analysis, protein-protein interaction, and protein conformational changes due to its enhanced brightness and small size. However, questions related to its mechanism of interaction with the substrate, furimazine, as well as bioluminescence activity remain elusive. Here, we combined molecular dynamics (MD) simulation and mutational analysis to show that the R162A mutation results in a decreased but stable bioluminescence activity of NLuc in living cells and in vitro. Specifically, we performed multiple, all-atom, explicit solvent MD simulations of the apo and furimazine-docked (holo) NLuc structures revealing differential dynamics of the protein in the absence and presence of the ligand. Further, analysis of trajectories for hydrogen bonds (H-bonds) formed between NLuc and furimazine revealed substantial H-bond interaction between R162 and Q32 residues. Mutation of the two residues in NLuc revealed a decreased but stable activity of the R162A, but not Q32A, mutant NLuc in live cell and in vitro assays performed using lysates prepared from cells expressing the proteins and with the furimazine substrate. In addition to highlighting the role of the R162 residue in NLuc activity, we believe that the mutant NLuc will find wide application in designing in vitro assays requiring extended monitoring of NLuc bioluminescence activity. SignificanceBioluminescence has been extensively utilized in developing a variety of biological and biomedical assays. In this regard, engineering of brighter bioluminescent proteins, i.e. luciferases, has played a significant role. This is acutely exemplified by the engineering of the NLuc luciferase, which is small in size and displays much enhanced bioluminescence and thermal stability compared to previously available luciferases. While enhanced bioluminescent activity is desirable in a multitude of biological and biomedical assays, it would also be useful to develop variants of the protein that display a prolonged bioluminescence activity. This is specifically relevant in designing assays that require bioluminescence for extended periods, such as in the case of biosensors designed for monitoring slow enzymatic or cellular signaling reactions, without necessitating multiple rounds of luciferase substrate addition or any specialized reagents that result in increased assay costs. In the current manuscript, we report a mutant NLuc that possesses a stable and prolonged bioluminescence activity, albeit lower than the wild-type NLuc, and envisage a wider application of the mutant NLuc in designing biosensors for monitoring slower biological and biomedical events.

MicroMPN: methods and software for high-throughput screening of microbe suppression in mixed populations.

We created a unified set of software and laboratory protocols for screening microbe libraries to assess the suppression of a pathogen in a mixed microbial community. Existing methods of fluorescent labeling were combined with the most probable number (MPN) assay in a microplate format to enumerate the reduction of a pathogenic soil microbe from complex soil matrices. This work provides a fluorescent expression vector available from Addgene, step-by-step laboratory protocols hosted by protocols.io, and MicroMPN, a command-line software for processing plate reader outputs. MicroMPN simplifies MPN estimation from 96- and 384-well microplates. The microplate screening assay is amenable to robotic automation with standard liquid handling robots, further reducing the hands-on processing time. This tool was designed to evaluate synthetic microbial communities for use as microbial inoculates or probiotics. The fluorMPN method is also useful for screening chemical and antimicrobial libraries for pathogen suppression in complex bacterial communities like soil.

Surfactant-Polymer Composition for Selective Water Shut-Off in Production Wells.

Today, a significant part of production wells' stock has a high water cut percentage of 90% and above. Obviously, for this reason, the need to develop new and improved existing technologies for water shut-off in wells increases every year. Physico-chemical methods of water shut-off are based on the application of special reagents and compositions that plug the pathways of water inflow to the well. Depending on the mechanism and specific features of water barrier formation, isolation methods are divided into selective and non-selective. This article investigates the possibility of using hydrolyzed polyacrylonitrile as a gel-forming and precipitation-forming reagent for water shut-off technologies in production wells. A surfactant-polymer composition for the isolation of water inflow in production wells in objects with high salinity in formation water, possessing physical and chemical selectivity and providing permeability reduction only in water-saturated intervals, is proposed. The developed composition is the invert emulsion, which makes it possible to carry out treatment at a distance from the well and solve the problem of possible premature gel formation directly in the wellbore. The lowest effective concentration of HPAN in an aqueous solution for use as a gel-forming and sedimentation reagent was determined experimentally (5.0 wt% and more). The interaction of the polymer solution with a chromium crosslinker allows obtaining structured gels in the whole volume of the system. The structure of the gels was evaluated using the Sydansk classifier with the assignment of a letter code from A to J. It was experimentally proved that the structure of the obtained gels depends on the temperature and content of the crosslinking agent in the system; the more crosslinking agent in the composition of the system, the stronger the structure of the resulting gel. The optimal ratio of polymer and crosslinking agent to obtain a strong gel was obtained, which amounted to 5:1 by weight of dry polymer powder. For the HPAN concentration of 5 wt% according to the Sydansk classifier, the gel structure had the code "H"-slightly deformable non-flowing gel. The dependence of the volume of gel sediment obtained because of the interaction with mineralized water on the polymer concentration was studied. It was proved that an increase in the concentration of hydrolyzed polyacrylonitrile in the solution, as well as an increase in the concentration of calcium ions in mineralized water, leads to a larger volume of the resulting gel or precipitate and to the strengthening of the gel structure. The results of rheological studies of the developed composition, as well as experiments on thermal stability, are presented. The results of filtration tests on bulk reservoir models demonstrated the selectivity of the developed composition. The obtained value of the residual resistance factor for the oil-saturated low-permeability model was 1.49 units; the value of the residual resistance factor for the water-saturated high-permeability model was 18.04 units. The ratio of the obtained values of the residual resistance factor, equal to 0.08 (much less than 1), can characterize the developed composition as a selective material for water shut-off in producing wells. Existing technologies for water shut-off based on HPAN do not allow for making a treatment at a distance from the well and require the use of technological solutions to prevent premature gel sedimentation in the well. The developed composition makes it possible to solve the problem of premature gelation. In addition, the composition can form a blocking screen in highly permeable water-saturated zones. The development can be useful for deposits with difficult conditions (high mineralization in reservoir waters, boreholes with a horizontal end, elevated temperatures up to 80 °C).

Liquid and Solid Hybridization Methods to Detect RNAs.

Northern blotting (NB) has been a long-standing method for RNA detection. However, its labor-intensive nature, reliance on high-quality RNA, and use of radioactivity have diminished its appeal over time. Nevertheless, the emergence of microRNAs (miRNAs) has reignited the demand for sensitive and quantitative NB techniques. We have recently developed cost-effective and rapid protocols for RNA detection using solid and liquid hybridization (LH) techniques which exhibit high sensitivity without the need for radioactive or specialized reagents like locked nucleic acid (LNA) probes. Our assays incorporate biotinylated probes and improved techniques for probe hybridization, transfer, cross-linking, and signal enhancement. We demonstrate that while NB is sensitive in detecting mRNAs and small RNAs, our LH protocol efficiently detects these as well as miRNAs at lower amounts of RNA, achieving highersensitivity comparable to radiolabeled probes. Compared to NB, LH offers benefits of speed, sensitivity, and specificity in detecting mRNAs, small RNAs, and miRNAs.

SUBJECT: REVIEW METHODS OF DETECTING SUGAR IN A SAMPLE

The sugar is a simple type of carbohydrate and represents the main source of energy to power the biological and physical activities in the body. However, it has been reported that over weight gain, imperfect metabolic processes and heart diseases are mainly related to excess sugar intake, which causes diabetic complains. This has raised the demand of having reliable test methods for quantifying sugar amount in different samples to both monitor sugar levels and to study the biological effects of carbohydrates. This provides valuable information in various fields, including food science, biochemistry, and medicine. Several test methods have been applied to detect the presence of sugar in a biological sample. Conventional tests imply using special chemical reagents. Two well known tests are discussed briefly in this report, namely Molich’s and Fehling's tests. The chemical reagents, test procedures, precaution as well as advantages and limitations were comprehensively discussed. A comparison between the two tests has been thoroughly addressed.

РАЗДЕЛЕНИЕ ВОДОНЕФТЯНЫХ ЭМУЛЬСИЙ ИЗ НЕФТЕЙ ВОСТОЧНО-СИБИРСКИХ МЕСТОРОЖДЕНИЙ

The article deals with the problem of the formation of oil-water emulsions. It is shown that some chemical compounds in crude oil - asphaltenes, resins, organic acids, chloride salts, mechani-cal impurities and solid paraffins have an emul–gating ability due to their surface activity. The regulatory requirements for crude and incoming oil for deep processing are analyzed. The data on the physical properties and component composition of the oils extracted in the fields of East-ern Siberia are given, on the basis of which it is concluded that these oils belong to the category of resinous and paraffin and can give stable emulsions in a mixture with water. For dehydration and desalination of oil, it is proposed to use special chemical reagents - demulsifiers. The meth-odology and results of an experimental comparison of the effectiveness of surfactants for the de-tection of a demulsifier suitable for the separation of oil-water emulsions from the oils of East Siberian fields are presented

Method for cryopreservation of trigeminal ganglion for establishing primary cultures of neurons and glia

BackgroundPrimary neuronal cultures are used to elucidate cellular and molecular mechanisms involved in disease pathology and modulation by pharmaceuticals and nutraceuticals, and to identify novel therapeutic targets. However, preparation of primary neuronal cultures from rodent embryos is labor-intensive, and it can be difficult to produce high-quality consistent cultures. To overcome these issues, cryopreservation can be used to obtain standardized, high-quality stocks of neuronal cultures. New methodIn this study, we present a simplified cryopreservation method for rodent primary trigeminal ganglion neurons and glia from Sprague-Dawley neonates, using a 90:10 (v/v) fetal bovine serum/dimethyl sulfoxide cell freezing medium. ResultsCryopreserved trigeminal ganglion cells stored for up to one year in liquid nitrogen exhibited similar neuronal and glial cell morphology to fresh cultures and retained high cell viability. Proteins implicated in inflammation and pain signaling were expressed in agreement with the reported subcellular localization. Additionally, both neurons and glial cells exhibited an increase in intracellular calcium levels in response to a depolarizing stimulus. Cryopreserved cells were also transiently transfected with reporter genes. Comparison with existing methodsOur method is simple, does not require special reagents or equipment, will save time and money, increase flexibility in study design, and produce consistent cultures. ConclusionsThis method for the preparation and cryopreservation of trigeminal ganglia results in primary cultures of neurons and glia similar in viability and morphology to fresh preparations that could be utilized for biochemical, cellular, and molecular studies, increase reproducibility, and save laboratory resources.

Carbon-Based Enzyme Mimetics for Electrochemical Biosensing.

Natural enzymes are used as special reagents for the preparation of electrochemical (bio)sensors due to their ability to catalyze processes, improving the selectivity of detection. However, some drawbacks, such as denaturation in harsh experimental conditions and their rapid de- gradation, as well as the high cost and difficulties in recycling them, restrict their practical applications. Nowadays, the use of artificial enzymes, mostly based on nanomaterials, mimicking the functions of natural products, has been growing. These so-called nanozymes present several advantages over natural enzymes, such as enhanced stability, low cost, easy production, and rapid activity. These outstanding features are responsible for their widespread use in areas such as catalysis, energy, imaging, sensing, or biomedicine. These materials can be divided into two main groups: metal and carbon-based nanozymes. The latter provides additional advantages compared to metal nanozymes, i.e., stable and tuneable activity and good biocompatibility, mimicking enzyme activities such as those of peroxidase, catalase, oxidase, superoxide dismutase, nuclease, or phosphatase. In this review article, we have focused on the use of carbon-based nanozymes for the preparation of electrochemical (bio)sensors. The main features of the most recent applications have been revised and illustrated with examples selected from the literature over the last four years (since 2020).

Issues of improving well construction processes and their environmental component

Purpose. Consistent analysis of complex issues of technical and technological components of the well construction cycle, taking into account the need to ensure the achievement of high technical and economic indicators while simultaneously complying with restrictive standards of environmental protection and technological safety. Research methodology. Theoretical and laboratory studies of the features of the use of special process fluids were carried out using modern methods of analytical analysis and experimental research, in particular through the use of general principles of mathematical and physical modeling, methods for processing research results in EXCEL, MATHCAD, instrumentation and materials. The flow of downhole circulation processes was simulated on experimental stands at the training drilling site of the Dnipro University of Technology using a ZIF-650M drilling rig and a UKB-4P drilling rig, as well as corresponding drilling and auxiliary tools. Research results. The key issues of organizing the construction cycle of wells for various purposes are considered. The features of the use of special technological fluids at different stages of the complex process of obtaining a reliable communication channel in a mountain range with a deep deposit, in the form of a well, have been studied. The essence of individual operations during flushing, securing and cementing a wellbore is analyzed. The results of studies of the technological properties of clay washing liquids treated with special chemical reagents are presented. Specific examples of approaches to ensuring the implementation of well technological operations are highlighted. The need to comply with environmental safety standards is substantiated. Originality. According to the purpose of the research, laboratory study methods have established the existence of a correlation between the concentration of surfactants (using the example of sulfonol and ditalane) and indicators of the technological properties of drilling fluids, namely viscosity and fluid loss. These characteristics change upward or downward, corresponding to the presence or absence of a background carbon-alkaline reagent. This changes the nature of the interaction of flushing fluids with the walls of the wellbore, and its positive effect is manifested in the effect of hydrophobization and minimal impact on the environment. Practical implications. The logical and consistent study of the influence of individual components of the flushing, cementing and cementing processes on the overall performance of well construction, presented in the work, allows us to evaluate potential directions for research to improve the development cycles of mineral deposits. The processes of the technological cycle of constructing wells for various purposes and other related works are considered on the basis of ensuring compliance with environmental protection standards.

COMPOUND CONTENT OF LOCAL CURCUMIN (CURCUMA XANTHORRHIZA) IN NORTH SULAWESI, INDONESIA

This Zingiberaceae plant is widely used as medicine, including temulawak. Because curcuma (C.xanthorrhiza) is a medicinal plant that has many benefits and includes plants needed in large quantities compared to other medicinal plants. Traditionally rhizomes Temulawak is used to treat stomach ailments, liver disorders, constipation, diarrhea, dysentery, fever, hemorrhoids, hypotriglyceridaemic, and anti-inflammatory. Study about plant Curcuma local ignite still not enough researched so that not yet get information compounds which contained in the local curcuma of North Sulawesi for the development for biopharmaceutical development, mapping in the distribution of plants, ecology, plant conservation and protection to maintain local wisdom. Histochemical Test is a method to determine the content of chemical compounds in a plant tissue qualitative. Testing can be done by adding a special reagent or solution to the incision organ plant and will give color which Specific. Activity study this will test curcumin compound group on local ginger from North Sulawesi by histochemical method. research that done is study non experiment with design descriptive qualitative. The results showed that North Sulawesi local white temulawak powder contained curcumin in a sample with a sample weight of 0.10 g at a sample spotting volume of 20 µl with a sample spotting volume of 2040 nanograms, curcumin levels were <0.10 nanograms/mg. Curcumin biosynthesis is influenced by site conditions, agro-climate, genotype, and plant cultivation.

Alternative probe hybridization buffers for target RNA depletion and viral sequence recovery in NGS for poultry samples

Non-targeted next generation sequencing (NGS) is widely applied to identify the diversity of pathogens in field samples. However, abundance of host RNA (especially rRNA) and other environmental nucleic acids can reduce the abundance of pathogen specific reads of interest, reduce depth of coverage and increase surveillance costs. We presently deplete chicken- and selected bacterial-specific rRNAs in poultry field RNA samples with complementary DNA probes in a commercially available probe hybridization buffer followed by digestion of the RNA:DNA hybrids with RNase H. Because the current buffer is an expensive special order reagent of proprietary composition, we tested in-house and other commercially available buffers and identified a viable alternative that yields equivalent host rRNA depletion and viral-specific reads in poultry samples as the current special order reagent but at a reduced cost.