Colorimetric Test Research Articles

Molecular test for COVID-19 diagnosis based on a colorimetric genomagnetic assay

The world is in a long pandemic period caused by the SARS-CoV-2 virus and massive diagnostic tests to assist efforts to control the spread of the disease and also to avoid new coronavirus variants are still needed. Herein, we propose a simple and accurate saliva-based colorimetric test for the diagnosis of COVID-19. Magnetic beads (MBs) modified with a sequence of single-strand DNA (ssDNA) complementary to the N gene of the SARS-CoV-2 RNA were developed and used for magnetic capture and separation from a complex saliva sample. A second biotinylated ssDNA sequence was applied, and the colorimetric detection was carried out by adding streptavidin-horseradish peroxidase conjugate, H2O2, and tetramethylbenzidine (TMB) as chromogenic substrate. The test does not require viral RNA isolation, transcription, or amplification steps and can be performed at room temperature. The molecular assay test can be run using 96-well microplates, allowing the diagnosis of a large number of samples in 90 min. A simple support for magnets was designed and constructed using a 3D printer that allows the magnetic separations directly in the 96-well microplate. The colorimetric test showed an excellent ability to discriminate between healthy individuals and patients infected with SARS-CoV-2, with 92% and 100% of clinical sensitivity and specificity, respectively. This performance was similar to that achieved using the gold standard RT-PCR technique. The proposed genomagnetic assay offers an opportunity to greatly increase population testing, contribute to controlling the spread of the virus, and improve health equity in testing for COVID-19.

Inhibition of NO Production in LPS-Stimulated Primary Rat Glial Cells by Gnidilatimonoein and Extract of Daphne mucronata.

In the CNS, glial cells are involved in neuroinflammation and neuropathic pain. The glial cells are activated by a variety of pathological conditions and release pro-inflammatory mediators, including nitric oxide (NO). Overexpression of iNOS (inducible nitric oxide synthase) and extra NO is detrimental to neurophysiology and neuronal viability. This study aimed to examine the effect of Gnidilatimonein isolated from D. mucronata and its leaves extract (as natural phytochemicals) on NO production in the LPS-induced primary glial cells. A preparative HPLC method was used to isolate gnidilatimonoein from leaves ethanolic extract. Various doses of Gnidilatimonoein, the ethanolic extract were applied to primary glial cells inflamed by lipopolysaccharide. A Colorimetric test, an MTT assay, and a RT-PCR analysis were then performed to analyze and compare NO production, cell viability, and iNOS expression. Gnidilatimonoein treatment of pretreated primary glial cells significantly inhibited iNOS expression and decreased NO synthesis. Plant extracts also reduced NO production in inflamed microglial and glial at 0.1-3 mg.mL-1. At these concentrations, none of these compounds exerted a cytotoxic effect, suggesting that their anti-inflammatory effects were not due to the death of cells. This study indicates that D. mucronata and its active compound, Gnidilatimonoein, could have restrained effects on the expression of iNOS on the induced glial cells; however, further investigation is warranted.

An Antioxidant and Intelligent pH-Sensitive Composite Film Based on Gelatin and Persian Gum Using Purple Carrot Extract

The present research is aimed at developing novel intelligent pH-sensitive films based on Persian gum and gelatin using purple carrot extract. To this end, films were developed by mixing polymer solutions containing Persian gum (5%) and gelatin (5%), in equal proportions and partially replacing distilled water of solutions with purple carrot extract at 0, 15, 30, and 45%. Total anthocyanin content, antioxidant activity, color indices, and tensile strength of films were evaluated. Purple carrot extract incorporation in the film caused a slight reduction in the lightness and an increase in the anthocyanin content, DPPH inhibition, yellowness, redness, and tensile strength. The sample containing 45% extract (anthocyanin concentration was 2.39 mg/ml) was selected as the best pH-sensitive film based on mechanical and antioxidant properties and evaluated in terms of moisture content, moisture absorption, pH, heat seal ability, thickness, contact angle, FTIR, X-ray diffraction, and colorimetric tests. The sensitivity of the film to pH changes was assessed in the range of 2-12. The color of the film changed from pink to blue as a function of pH. The results proved that films based on Persian gum, gelatin, and purple carrot extract could be used as novel food grade biomaterials to monitor the freshness/spoilage of food.

Evaluation of tributyrin supplementation in milk replacer on diarrhoea occurrence in preweaning Holstein calves

Neonatal calf diarrhoea is one of the most important health challenges in cattle herds causing substantial economic losses and antimicrobial use. Due to the raising problem of antimicrobial resistance, effective alternatives are urgently required, in line with European policies. The aim of this study was to evaluate the effect of tributyrin supplementation in milk replacer on diarrhoea, performance and metabolic status in preweaning Holstein calves. Twelve newborn calves, after colostrum administration, were randomly allotted in two experimental groups for 42 days: control (CTRL) fed milk replacer, tributyrin (TRIB) fed milk replacer supplemented with 0.3% of liquid tributyrin on milk powder weight. Calves BW was recorded on a weekly basis from day 7 to day 42, and feed intake was recorded daily to calculate zootechnical performance. Faecal consistency was assessed daily through the faecal score (0–3 scale; considering diarrhoea moderate = 2 and severe = 3). Faecal samples were collected weekly from rectal ampulla for microbiological analysis by plate counting method evaluating the number of total bacteria, lactic acid bacteria and coliform bacteria. On day 0 and day 42, individual blood samples were collected from jugular vein for metabolic profile analysis. Serum samples of day 42 were also evaluated for the antioxidant barrier using a colorimetric test, while glucagon-like peptide 2 and diamine oxidase concentrations were measured through immunoenzymatic assays. Tributyrin supplementation did not influence the zootechnical performance of calves over 42 days of trial. Diarrhoea frequency was significantly lower in TRIB compared to CTRL group (27.91 and 38.37%; P < 0.01) considering the whole experimental period. In particular, the major effect was observed for moderate diarrhoea in TRIB group that showed a significantly reduced frequency compared to CTRL (P < 0.01) thus suggesting a preventive effect of tributyrin. Faecal total bacterial, lactic acid and coliform bacteria counts did not show differences between groups. Urea serum concentrations tended to be lower in TRIB compared to CTRL, indicating an efficient utilisation of dietary protein. Antioxidant barrier and glucagon-like peptide 2 were comparable between CTRL and TRIB on day 42. Diamine oxidase concentrations were significantly decreased in TRIB compared to CTRL group after 42 days of trial (P < 0.01), suggesting a higher gut epithelial integrity probably due to lower diarrhoea frequency and the nourish effect of tributyrin on enterocytes. In conclusion, tributyrin could be considered as a valuable bioactive feed additive to decrease the neonatal diarrhoea occurrence and support intestinal integrity in preweaning calves.

A Novel RT-LAMP for the Detection of Different Genotypes of Crimean-Congo Haemorrhagic Fever Virus in Patients from Spain.

Crimean-Congo haemorrhagic fever (CCHF) is a potentially lethal tick-borne viral disease with a wide distribution. In Spain, 12 human cases of CCHF have been confirmed, with four deaths. The diagnosis of CCHF is hampered by the nonspecific symptoms, the high genetic diversity of CCHFV, and the biosafety requirements to manage the virus. RT-qPCR and serological tests are used for diagnosis with limitations. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) could be an effective alternative in the diagnosis of the disease. However, none of the few RT-LAMP assays developed to date has detected different CCHFV genotypes. Here, we designed a RT-LAMP using a degenerate primer set to compensate for the variability of the CCHFV target sequence. RT-LAMP was performed in colorimetric and real-time tests on RT-qPCR-confirmed CCHF patient samples notified in Spain in 2020 and 2021. Urine from an inpatient was analysed by RT-LAMP for the first time and compared with RT-qPCR. The amplicons obtained by RT-qPCR were sequenced and African III and European V genotypes were identified. RT-LAMP amplified both genotypes and was more sensitive than RT-qPCR in urine samples. We have developed a novel, rapid, specific, and sensitive RT-LAMP test that allows the detection of different CCHFV genotypes in clinical samples. This pan-CCHFV RT-LAMP detected viral RNA for the first time in urine samples. It can be easily performed as a single-tube isothermal colorimetric method on a portable platform in real time and without the need for expensive equipment, thus bringing molecular diagnostics closer to rural or resource-poor areas, where CCHF usually occurs.

Implementation of orthogonal subspace projection on ASTER data for exploring phosphorite -a study in Chhatarpur, Madhya Pradesh, India

In this study, we have identified surface exposures of phosphorite using visible near infrared (VNIR) and shortwave infrared (SWIR) bands of Advanced space borne thermal emission and reflection radiometer (ASTER) data. We analysed the spectral contrast of dolomite and phosphorite samples collected from the ground and also from the pixels of the ASTER image composite. We observed the consistent spectral contrast between the samples of dolomite and phosphorite and used the same for delineating phosphorite within the dolomite and associated carbonate rocks. In this regard, we first delineated dolomite and associated carbonate rocks using feature oriented principal component of the iron rich carbonate rocks. Subsequently, we implemented partial spectral unmixing method, known as orthogonal subspace projection on ASTER VNIR and SWIR bands to delineate phosphorite within dolomite. We could trace a few new occurrences of phosphorite in the study area beyond the known occurrence at Hirapur open cast mining area. We have validated the identified new areas during field survey using a rapid colorimetric test for phosphate to ascertain the exploration importance of these targets. We further estimated P2O5 grade content of the phosphorite samples in these sites and found few of them have very high P2O5 content.

Phytochemical Screening and Antioxidant Activity of Selected Estonian Galium Species.

The aim of the present study was to examine three different Galium species from the native population of Estonia, Galium verum, Galium aparine, and Galium mollugo, to characterise their non-volatile and volatile phytochemical composition and antioxidant activity. The main groups of bioactive compounds in the plants were quantified by colorimetric tests, showing high concentrations of polyphenols (up to 27.2 ± 1.5 mg GAE/g), flavonoids (up to 7.3 ± 0.5 mg QE/g) and iridoids (up to 40.8 ± 2.9 mg AE/g). The species were compared using HPLC-DAD-MS/MS, revealing some key differences in the phytochemical makeup of the extracts. The most abundant compound in the extracts of Galium verum blossoms and herb was found to be asperuloside, in Galium aparine herb, asperulosidic acid, and in Galium mollugo herb, chlorogenic acid. Additionally, the composition of volatile compounds was analysed by SPME-GC-MS. The degree of variability between the samples was high, but three volatiles, hexanal, anethole, and β-caryophyllene, were quantified (≥1%) in all analysed samples. The antioxidative activity of all extracts was evaluated using the ORACFL method, demonstrating that the Galium species from Estonia all exhibit strong antioxidant capacity (up to 9.3 ± 1.2 mg TE/g). Out of the extracts studied, Galium verum blossoms contained the highest amounts of bioactives and had the strongest antioxidant capacity.

Comparisons between Hydro and Steam Distillation Processes to Extract Prunella Vulgaris Volatile Compounds, and their Anti-oxidative Activities

Objective The aim is to verify our earlier suggestion which accounted for the dynamics of volatile compounds extraction from the herb Prunella vulgaris (PV) using steam distillation. Then, the anti-oxidative property of PV is explored.&#x0D; Methods Because we suggested earlier that the inefficient extraction using steam distillation was due to the obstruction of a mass of herb in the steam flow path, we used hydro distillation which tried to eliminate this obstacle. We used GC-MS to characterize the volatile compounds extracted, and thus compare the extraction efficiency. Then, treating the cancer cells from the SCC154 cell-line with the distillate, the cancer cell cytotoxicity was assessed using the colorimetric test reagent, the Cell Counting Kit-8. To assess the anti-oxidative activity of the PV distillate, Folin-Ciocalteu reagent was used.&#x0D; Results We successfully showed that the removal of the obstacle, formed by the mass of herb in the steam flow path, enhanced the efficiency of volatile compound extraction. Also, we showed that the PV distillate did not exhibit anti-oxidative activity.&#x0D; Conclusions Hydro distillation is a more efficient method than steam distillation to extract volatile compounds from the PV herb. However, mild heating did not provide sufficient energy to the convection of the boiling water to move the floating herb; the obstacle still existed and limited the efficiency of extraction also. For another issue of the antioxidant effect of the volatile compounds from PV, it was studied using the Folin-Ciocalteu reagent. It showed that the PV volatile compounds did not possess antioxidant property.&#x0D; Keywords: Prunella vulgaris, Xia Ku Cao, Chinese Medicine, steam distillation, hydro distillation, anti-oxidative effect

Construction of a flame retardant polyurethane elastomer with degradability, high mechanical strength and shape memory

In order to promote the application scene of polyurethane (PU) elastomers, the low degradability and poor fire safety are the key problems that need to be solved urgently. In this work, the polyurethane elastomers (PCL-HDI-THPOx) with flame retardant and degradable groups were synthesized. In the Microscale combustion colorimeter (MCC) test, the Total Heat Release (THR) and peak of Heat Release Rate (pHRR) of were decreased by 24% and 26%, respectively. Due to the enhancement of the internal crosslinking network density of the elastomer by THPO, the tensile strength and toughness increased by 450% and 117%, respectively. Differential Scanning Calorimetry (DSC) results showed the glass transition temperature (Tg) was between 30 °C and 40 °C, showing good shape memory properties. Surprisingly, the novel elastomer completely degraded in ethanol at 60 °C. Its excellent comprehensive performance will be welcomed in the fields of wearable electronic devices and load-bearing materials.

Lanthanum-doped carbon quantum dots (La-CQDs) for detection of Fe3+ in colorimetric test paper and information anti-counterfeiting

Based on the fluorescence quenching principle, a novel lanthanum-doped carbon quantum dots (La-CQDs) fluorescence probe was successfully developed for ion detection of Fe3+. La-CQDs with a fluorescence quantum yield of 29.93% was prepared in ethanol solution using ammonium citrate as the carbon source and lanthanum nitrate as the dopant. The luminescence performance of La-CQDs was enhanced by La doping and maintained over 90% of the maximum fluorescence intensity in different anions, water content and organic solvent environments. In the range of 25–200 μM, the fluorescence intensity of La-CQDs displayed a good linear relationship (Y = 0.00675X+ 1.07679, R2 = 0.9888) with the concentration of Fe3+ and its limit of detection (LOD) was calculated to be 6.78 μM. The method was successfully applied to the detection of Fe3+ in green tea and the measurement error was less than 3%. In addition, La-CQDs was developed for fluorescent colorimetric test paper and information anti-counterfeiting, while the fluorescence quenching effect was observed intuitively on the paper.

A Fluorescent and Colorimetric Chemosensor Detecting Pd2+ Based on Chalcone Structure with Triphenylamine.

A fluorometric and colorimetric chemosensor DiPP ((E)-3-(4-(diphenylamino)phenyl)-1-(pyridin-2-yl)prop-2-en-1-one) based on chalcone structure with a triphenylamine group was synthesized. Sensor DiPP detected Pd2+ with fluorescence turn-off and via colorimetry variation of yellow to purple. The binding ratio of DiPP to Pd2+ turned out to be 1 : 1. Detection limits for Pd2+ by DiPP were analyzed to be 0.67 µM and 0.80 µM through the fluorescent and colorimetric methods. Additionally, the fluorescent and colorimetric test strips were applied for probing Pd2+ and displayed that DiPP could obviously discriminate Pd2+ from other metals. The binding feature of DiPP to Pd2+ was presented by ESI-mass, Job plot, NMR titration, ESI-mass, and DFT calculations.

Debugging periodate oxidation of cellulose: Why following the common protocol of quenching excess periodate with glycol is a bad idea

Periodate oxidation of cellulose to produce “dialdehyde cellulose” (DAC) has lately received increasing attention in sustainable materials development. Despite the longstanding research interest and numerous reported studies, there is still an enormous variation in the proposed preparation and work-up protocols. This apparently reduces comparability and causes reproducibility problems in DAC research. Two simple but prevalent work-up protocols, namely glycol quenching and filtration/washing, were critically examined and compared, resulting in this cautionary note. Various analytical techniques were applied to quantify residual iodine species and organic contaminations from quenching side reactions. The commonly practiced glycol addition cannot remove all oxidising iodine compounds. Both glycol and the formed formaldehyde are incorporated into DAC's polymeric structure. Quenching of excess periodate with glycol can thus clearly be discouraged. Instead, simple washing protocols are recommended which do not bear the risk of side reactions with organic contaminants. While simple washing was sufficient for mildly oxidised celluloses, higher oxidised samples were more likely to trap residual (per)iodate, as determined by thiosulfate titration. For work-up, simple washing with water is proposed while determining potential iodine contaminations after washing with a simple colorimetric test and, if needed, removal of residual periodate by washing with an aqueous sodium thiosulfate solution.

Assessment of the Physico-Chemical Conformity of an Antiseptic Solution: Case of Polyvidone - Iodine 10%

Introduction: Antiseptics have been used for many years in human and veterinary medicine. Over the years, the lengthening of surgical procedures in human surgery and the increasingly frequent use of inert material has increased the risk of infections. This was why skin antisepsis has become very important in the prevention of infections of the surgical wound and resulting complications. In Mali, at the Gabriel Touré University Hospital, the most prescribed family of antiseptics was the halogenated ones, in particular 10% povidone iodine for wound and surgical site antisepsis. This study was aimed to control the quality of 10% povidone iodine used in the hospital. Methodology: We conducted at the Medicines Quality Control Service of the National Health Laboratory of Mali (LNS) a retrospective analytical study on the quality control of 10% povidone iodine. We obtained 10% povidone iodine samples from the Pharmacie Populaire du Mali (PPM). We analyzed 48 batches of samples in regards of visual inspection, average volume, titrimetry, colorimetric test and pH control. Results: All 48 analyzed batches were compliant, i.e. contained the active ingredient and could be used in health facilities or at home as an antiseptic. Conclusion: All analyzed batches of 10% povidone iodine samples did not show any cases of inconsistency and therefore could be used in the management of wound dressings. Future Direction: Use other analytical methods like HPLC to test the quality control of povidone and other types of antiseptics.&#x0D; Keywords: antiseptic, povidone-iodine 10%, compliance, quality control

Functional Olive Pit Powders: The Role of the Bio-Based Filler in Reducing the Water Uptake Phenomena of the Waterborne Paint

In this study, olive pit powders were added to a polyurethane-acrylate paint for examining the impact of two alternative functionalization processes in increasing the filler hydrophobicity in an effort to increase the durability of the paint. In order to look into potential changes in morphology and appearance owing to the surface conversion treatments of the two bio-based additives, the coatings were examined using electron microscopy and colorimetric tests. The coating’s resilience and the hydrophobic/hydrophilic role of the fillers were evaluated by salt spray chamber exposure, contact angle measurements, paint liquid resistance, UV-B exposure, and electrochemical impedance spectroscopy measurements, which highlighted the reduction in water absorption inclination of the filler made of lignocellulose due to the silane and wax functionalization. This study demonstrated that the bio-based filler, if properly functionalized, can actually be implemented as multifunctional pigment in waterborne paints, giving specific aesthetic characteristics, but also improving the barrier performance of the polymeric matrix and increasing the durability of the composite coating.

Lack of Association between YEASTONE Antifungal Susceptibility Tests and Clinical Outcomes of Cryptococcus Meningitis.

The relation between antifungal susceptibility and treatment outcomes is not well-characterized. There is paucity of surveillance data for cerebrospinal fluid (CSF) isolates of cryptococcus investigated with YEASTONE colorimetric broth microdilution susceptibility testing. A retrospective study of laboratory-confirmed cryptococcus meningitis (CM) patients was conducted. The antifungal susceptibility of CSF isolates was determined using YEASTONE colorimetric broth microdilution. Clinical parameters, CSF laboratory indices, and antifungal susceptibility results were analyzed to identify risk factors for mortality. High rates of resistance to fluconazole and flucytosine were observed in this cohort. Voriconazole had the lowest MIC (0.06 µg/mL) and lowest rate of resistance (3.8%). In a univariate analysis, hematological malignancy, concurrent cryptococcemia, high Sequential Organ Failure Assessment (SOFA) score, low Glasgow coma scale (GCS) score, low CSF glucose level, high CSF cryptococcal antigen titer, and high serum cryptococcal antigen burden were associated with mortality. In a multivariate analysis, meningitis with concurrent cryptococcemia, GCS score, and high CSF cryptococcus burden, were independent predictors of poor prognosis. Both early and late mortality rates were not significantly different between CM wild type and non-wild type species.

Visual, customizable wood-based colorimetric test paper encapsulated with fluorescent carbon dots for rapid explosive detection

Wood research has recently enjoyed a renaissance due to its emerging application in various fields. This is due to the natural reaction platform and hierarchical porous structure of wood, which allows it to combine perfectly with functional materials, especially nanoscale materials. Here, we report a portable fluorescent wood (FW) test paper based on the combination of wood and carbon dots (CDs) for rapid identification and detection of nitroaromatic explosives. Thereinto, the CDs optimize the assembly of wood and open up new functions, and the naturally porous structure of wood provides grid management for CDs, effectively avoiding the accumulation of CDs. Meanwhile, the CDs are immobilized in the wood skeleton via the multiple-blockades, and the fluorescence intensity is unaffected even under harsh environmental conditions, which ensures the FW test paper exhibits high sensitivity and selectivity for trinitrophenol. The quenching reached 100% by dropping 80 μM trinitrophenol with the ultrathin FW test paper (1 × 1 × 0.009 cm), and it is still significant in the presence of interference. Interestingly, the detection results can be monitored in real time with the naked eye. As far as we know, this is the first report of trinitrophenol detection with FW test paper.

Red cabbage extract-mediated colorimetric sensor for swift, sensitive and economic detection of urease-positive bacteria by naked eye and Smartphone platform

The bacterial pathogens have caused various serious infectious diseases in the human body, and even some threats to human life by leading to deaths. Enterobacteriaceae species especially urease positive ones, Proteus mirabilis (P. mirabilis) and Klebsiella pneumoniae (K. pneumoniae), show resistance to antibiotics and cause respiratory and urinary tract infections. We have developed natural indicator-incorporated colorimetric urease tests with a naked eye and smartphone readout to rapidly, sensitively and economically detect P. mirabilis and K. pneumoniae. We utilized anthocyanin found as a predominant component in red cabbage (Brassica oleracea) extract as a natural pH indicator instead of toxic and synthetic indicators. As a mechanistic explanation for the detection of P. mirabilis and K. pneumoniae, urease enzymes secreted from the P. mirabilis and K. pneumoniae hydrolyze urea to produce ammonia (NH3), which increases the pH value of the reaction environment and leads to deprotonation from anthocyanins. The changes in the molecular structure and electronic structure of anthocyanins are responsible for revealing many different colors. We demonstrated how some reaction parameters including the concentration of the bacteria (colony-forming unit, CFU), the concentration of anthocyanin in the tests, initial color and pH values (pHs) of the tests influence their detection performance. We further developed a 3D-printed smartphone platform with smartphone based digital image processing software to improve the detection limit and shorten the detection time. We claim that natural indicator-incorporated rapid urease tests providing colorimetric readout evaluated by the human eye and smartphone imaging processing has great potential in practical use and they can be implemented in clinics.

New Psychoactive Substances: Major Groups, Laboratory Testing Challenges, Public Health Concerns, and Community-Based Solutions

Across communities worldwide, various new psychoactive substances (NPSs) continue to emerge, which worsens the challenges to global mental health, drug rules, and public health risks, as well as combats their usage. Specifically, the vast number of NPSs that are currently available, coupled with the rate at which new ones emerge worldwide, increasingly challenges both forensic and clinical testing strategies. The well-established NPS detection techniques include immunoassays, colorimetric tests, mass spectrometric techniques, chromatographic techniques, and hyphenated types. Nonetheless, mitigating drug abuse and NPS usage is achievable through extensive community-based initiatives, with increased focus on harm reduction. Clinically validated and reliable testing of NPS from human samples, along with community-driven solution, such as harm reduction, will be of great importance, especially in combating their prevalence and the use of other illicit synthetic substances. There is a need for continued literature synthesis to reiterate the importance of NPS, given the continuous emergence of illicit substances in the recent years. All these are discussed in this overview, as we performed another look into NPS, from differentiating the major groups and identifying with laboratory testing challenges to community-based initiatives.

Evaluation of two colorimetric microplate microtiter tests for susceptibility testing of clinical isolates of Mycobacterium tuberculosis to first-line antituberculosis drugs.

Drug-resistance tuberculosis (TB) is one of the most important global public health problems. Accurate and rapid drug-susceptibility testing is critical for the effective treatment of TB patients. Various colorimetric methods are used for anti-TB drug-susceptibility testing (DST) and minimum inhibitory concentration (MIC) determination. This study was conducted to evaluate the resazurin microtiter assay (REMA) and malachite green decolorization assay (MGDA). A total of 65 Mycobacterium tuberculosis strains isolated from patients with suspected TB using REMA and malachite green microtiter assay methods were tested against streptomycin (SM), isoniazid (INH), rifampicin (RIF), and ethambutol (ETB). The Mycobacterial Growth Indicator Tube 960 DST method was accepted as the gold standard in the evaluation of test results. The sensitivity of REMA and MGDA tests was found to be 87.5% and 62.5% for INH, respectively. RIF and SM sensitivity for both tests was 100%. While ETB sensitivity was 81.8 for the REMA test, this rate was 60% for the MGDA test. Specificity of both tests varied between 92.5% and 98.2% according to the drug types. REMA and MGDA are a simple, rapid, and low cost. They can be used as an alternative test for drug-susceptibility testing and MIC determination. Extensive studies and standardization are needed for the routine use of both tests.

Novel indole retinoid derivative induces apoptosis and cell cycle arrest and modulates AKT and ERK signaling in HL-60 cells.

Chemotherapy with targeted drugs is the first line therapy option for acute and chronic myeloid leukemia. However, hematopoietic stem cell transplantation may be used in high-risk patients or patients with failed responses to chemo drugs. Discovery and development of more effective new agents with lower side effects is the main aim of leukemia treatment. In this study, a novel retinoid compound with tetrahydronaphthalene ring was synthesized and evaluated for anticancer activity in human chronic and acute myeloid leukemia cell lines K562 and HL-60. Novel N-(1H-indol-1-yl)-5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalene-2-carboxamide was synthesized based on molecular hybridization of the two different bioactive structures retinoid head and indole. The effects of the synthesized carboxamide compound, which was referred to as compound 5, were determined in K562 chronic myeloid leukemia and HL-60 acute myeloid leukemia cell lines and L929 fibroblast cell line, which served as a control. Colorimetric MTT and caspase3 activity tests, flow cytometry, western blot, and microscopic examinations were used to evaluate biological activity. Compound 5 more effectively induced cell death in HL60 cells in comparison to K562 cells and L929 fibroblast cells. Therefore, further mechanism of cell death was investigated in HL60 cell line. It was found that compound 5 induced remarkable cytotoxicity, caspase3 activation, and PARP fragmentation in HL60 cells. Flow cytometric staining showed that the percentage of cells arrested in G0/G1 was also increased with compound 5 treatment. Important modulator proteins of cell proliferation p-ERK, p-AKT, and p-m-TOR were also found to be inhibited with compound 5 treatment. Collectively, our results reveal compound 5, which is a novel indole retinoid compound as a potential active agent for the treatment of acute promyelocytic leukemia.