RESUMEN

Approximately 7% of patients with newly diagnosed multiple myeloma (MM) experience bleeding complications with varying causes, but few reports have described these complications. Here we report the case of a patient with newly diagnosed MM who presented with a bleeding tendency and various coagulation abnormalities. Chromogenic assays, thrombin time, and reptilase time revealed the presence of a thrombin-inhibiting substance that inhibited release of fibrinopeptide A from fibrinogen. The coagulation abnormalities improved after treatment with daratumumab, lenalidomide, and dexamethasone. As the thrombin inhibition mechanism remains unclear, no previous studies have reported recent treatment outcomes in older patients producing thrombin-inhibiting substances, which can hinder clinical treatment. Therefore, we believe that the diagnosis and the treatment course of this case provide valuable information. Moreover, such case reports provide significant insights into the pathophysiology of bleeding complications associated with MM.

RESUMEN

Our aim was to compare the performance of complementary clinical laboratory approaches to monitoring exposure to apixaban and rivaroxaban, the most prescribed direct-acting oral anticoagulants (DOAC's): an automated commercial anti-Xa chromogenic assay suitable for emergency and pre-surgery testing and a laboratory-developed liquid chromatography-tandem mass spectrometry (LC-MS/MS) method employed for non-emergency analysis in plasma and in dried blood volumetric absorptive microsamples (VAMS) collectible by the patients in their homes. The full validation of the LC-MS/MS method was performed. Cross-validation of the methodologies was accomplished by processing 60 specimens collected for whole blood count and DOAC monitoring in a central clinical laboratory. For VAMS samples, dried plasma and whole blood calibrators were found to be suitable, and a cycle run for seven days could be implemented for rational and economic sample processing. The anti-Xa chromogrenic assay and the LC-MS/MS method delivered discordant plasma analyte concentrations. Moreover, the lack of agreement between plasma and VAMS concentrations was observed. Clinical laboratories must be aware of the differences between the performance of apixaban and rivaroxaban LC-MS/MS and anti-Xa assays. Hematocrit must always be measured along with VAMS samples to obtain accurate results.

RESUMEN

Timely and accurate identification of yeasts is essential for adequate treatment, considering the increase in antifungal resistance of some species, particularly for C. auris. Current matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) manufacturer's protocol for identification of yeasts requires 24- to 72-h cultivation on Sabouraud dextrose media (SAB), but not some of the mainstay primary culture media used in mycology such as inhibitory mold agar (IMA), Mycosel, CHROMagar Candida Plus, and CHROMagar Candida. As culture media can influence MALDI-TOF MS identification results, this study evaluated the accuracy and performance of identification of clinically relevant yeasts on these first-line media using the VITEK-MS MALDI-TOF MS system.IMPORTANCEIn this study, a panel of 140 strains (21 species) was used to assess the performance of the selected media. Although not in the manufacturer's list of accepted media, IMA and chromogenic media are suitable for the identification of yeasts on the VITEK-MS systems. CHROMagar Candida Plus allowed the identification of 135/140 isolates tested after 24-h incubation similar to SAB reference media (137/140). Yeast isolates that grew on Mycosel selective media were also reliably identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. VITEK-MS system with IVD database V3.2 correctly identified C. auris strains to the species level on CHROMagar Candida Plus alleviating the need for subcultivation and reduced turnaround time (24-72 h) to identification for patient screening.

RESUMEN

Background Vulvovaginal candidiasis (VVC) is a common fungal infection caused by an overgrowth of Candida species, primarily Candida albicans (C. albicans). Using HiCrome agar and tetrazolium reduction medium offers cost-effectiveness in Candida detection by eliminating the need for additional tests, reducing equipment costs compared to automated systems, and simplifying workflow with direct species identification while maintaining high specificity. They expedite detection by directly identifying Candida species based on colony colour, bypassing the multiple steps of phenotypic methods. This efficiency saves time in the laboratory, providing rapid results without the extended processing times associated with automated systems and facilitating prompt diagnosis and treatment decisions. These diagnostic tools are especially valuable in low-resource environments where a quick and accurate diagnosis of VVC is crucial for effective treatment and management of antifungal resistance. Aims and objectives This study aims to evaluate the efficacy of HiCrome agar and tetrazolium reduction medium's efficacy in speciating Candida species in VVC cases. Materials and methods A cross-sectional observational study was conducted at Saveetha Medical College and Hospitals, Chennai, India, over six months. High vaginal swabs from 126 patients suspected of VVC were collected and plated on Sabouraud dextrose agar (SDA), HiCrome Candida differential agar (Himedia, Mumbai, India), and tetrazolium reduction medium. The results were compared with those obtained from the VITEK2 compact system (bioMérieux, Marcy-l'Étoile, France). Results Of the 126 samples, 74.6% showed single yeast infections, 7.9% displayed mixed yeast infections, and 17.5% showed no growth. A total of 114 Candida isolates were identified. Both HiCrome agar and tetrazolium reduction medium accurately identified all isolates, with complete concordance with the VITEK2 compact system. The most commonly isolated species were C. albicans (55.2%), Candida tropicalis (32.4%), Candida glabrata (8.8%), and Candida parapsilosis (3.6%). Both media provided rapid and accurate presumptive identification in low-resource settings. Conclusions HiCrome agar and tetrazolium reduction medium demonstrated high sensitivity and specificity in identifying Candida species. These methods are reliable for rapid and accurate diagnosis, particularly in resource-limited settings. However, they may require supplementary tests for definitive species identification. The adoption of these diagnostic tools represents a significant advancement in clinical microbiology, improving VVC management and addressing antifungal resistance.

RESUMEN

PURPOSE: To examine the color stability of 3D-printed and milled, interim, and definitive, restorative materials after immersion in artificial saliva and wine for 1, 3, and 6 months. MATERIAL AND METHODS: The study used a 2 × 5 factorial design with 10 subgroups, including 2 immersion liquids (artificial saliva and wine) and 5 manufacturing technology and restorative material combinations (n = 10). Color measurements were taken using a contact-type digital spectrophotometer (CM-2600d Spectrophotometer; Konica Minolta Healthcare Americas Inc) before immersion and at 1 month (T1), 3 months (T3), and 6 months (T6) after immersion. The CIE2000 system was used to calculate quantitative measurements of color differences in ΔE00, and comparisons were made to the acceptability threshold (AT) and perceptibility threshold (PT). Repeated measures of ANOVA (α = 0.05) were used to compare differences in color changes between manufacturing technology/restorative material-immersion liquid combinations at T1, T3, and T6. RESULTS: To compare the effect of immersion liquid and time on the manufacturing technology/restorative material groups, the ΔE00 values were compared to the PT of 0.8 and the AT of 1.8. Wine caused significant color changes in ΔE00 values beyond the PT and AT values in all groups at all time intervals, except for the AT value of milled definitive crowns (hybrid nano-ceramic material). Wine immersion caused significant ΔE00 for all manufacturing technology/restorative material groups at all time intervals (1 month, 3 months, and 6 months) when compared to artificial saliva immersion (all p < 0.001). CONCLUSION: Upon exposure to artificial saliva, 80%-100% of samples from all groups remained within the acceptable and perceptible color change thresholds. The wine had significant chromogenic effects on all tested restorative materials, however, the milled definitive crowns (hybrid nano-ceramic material) showed the greatest color stability. For patients with heavy wine consumption, 3D-printed definitive crowns (hybrid ceramic-filled material) may show discoloration exceeding acceptable and perceptible color change limits.

RESUMEN

Sodium alginate (SA) is a marine polysaccharide biomass material that is environmentally friendly and exhibits color-changing properties under certain conditions. In this study, we have discovered sodium alginate solution to be chromogenic under four conditions, namely alkali-chromogenic, thermo-chromogenic, force-chromogenic and photo-chromogenic. Under simple strong alkaline conditions, sodium alginate forms clusters of blue light-absorbing chromogenic aggregates, which exhibit a bright yellow color at a certain size. Under different temperature conditions, SA shows varying shades of yellow, and the color tends to stabilize after 48 h of resting. The aggregates can be dispersed by stirring, which changes SA from yellow to colorless. The yellow color can then be recovered after resting. Additionally, exposure to sunlight can cause the yellow SA to fade, but the color can be restored by reheating. Therefore, the force-chromogenic and photo-chromogenic properties are reversible. This makes it a promising material for use in color-developing and indicating materials. It is expected to become a sodium alginate cluster pigment with broad application prospects in the future.

RESUMEN

This chapter describes how to test different amyloid preparations for catalytic properties. We describe how to express, purify, prepare and test two types of pathological amyloid (tau and α-synuclein) and two functional amyloid proteins, namely CsgA from Escherichia coli and FapC from Pseudomonas. We therefore preface the methods section with an introduction to these two examples of functional amyloid and their remarkable structural and kinetic properties and high physical stability, which renders them very attractive for a range of nanotechnological designs, both for structural, medical and catalytic purposes. The simplicity and high surface exposure of the CsgA amyloid is particularly useful for the introduction of new functional properties and we therefore provide a computational protocol to graft active sites from an enzyme of interest into the amyloid structure. We hope that the methods described will inspire other researchers to explore the remarkable opportunities provided by bacterial functional amyloid in biotechnology.

Asunto(s)

Amiloide , Proteínas de Escherichia coli , Escherichia coli , Ingeniería de Proteínas , alfa-Sinucleína , Proteínas tau , Amiloide/química , Amiloide/metabolismo , Proteínas de Escherichia coli/metabolismo , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , alfa-Sinucleína/química , alfa-Sinucleína/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Ingeniería de Proteínas/métodos , Proteínas tau/metabolismo , Proteínas tau/química , Humanos , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Pseudomonas/metabolismo , Pseudomonas/química , Catálisis , Dominio Catalítico

RESUMEN

Diagnostic laboratories in Aotearoa, New Zealand (NZ) refer cultures from faecal samples positive for Shiga toxin genes to the national Enteric Reference Laboratory for isolation of Shiga toxin-producing Escherichia coli (STEC) for epidemiological typing. As there was variation in the culture media being referred, a panel of 75 clinical isolates of STEC, representing 28 different serotypes, was used to assess six commercially available media and provide guidance to clinical laboratories. Recommendations were subsequently tested for a 3-month period, where STEC isolations and confirmations were assessed by whole genome sequencing analysis against the culture media referred. CHROMagar™ STEC (CH-STEC; CHROMagar Microbiology, Paris, France) or CH-STEC plus cefixime-tellurite sorbitol MacConkey agar was confirmed inferior to CH-STEC plus blood agar with vancomycin, cefsulodin, and cefixime (BVCC). The former resulted in fewer STEC types (n = 18) being confirmed compared to those from a combination of CH-STEC and BVCC (n = 42). A significant (P < .05) association with an STEC's ability to grow on CH-STEC and the presence of the ter gene cluster, and eae was observed. Culturing screen positive STEC samples onto both CH-STEC and BVCC ensures a consistently higher recovery of STEC from all clinical samples in NZ than CH-STEC alone.

Asunto(s)

Infecciones por Escherichia coli , Proteínas de Escherichia coli , Escherichia coli Shiga-Toxigénica , Humanos , Escherichia coli Shiga-Toxigénica/genética , Cefixima , Agar , Nueva Zelanda , Medios de Cultivo , Vancomicina , Cefsulodina , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/genética

RESUMEN

The capacity differences of seven catechin monomers to produce colors after treating with catechin-free extract were investigated. After 240-min reaction, only (-)-epicatechin (EC) and (+)-catechin (C) presented obvious luminous red color with L* values of 63.32-71.73, a* values of 37.13-46.44, and b* values of 65.64-69.99. Meanwhile, the decrease rate of EC and C was 43.52 %-50.35 %, which were significantly lower than those of other catechin monomers (85.91 %-100 %). The oxidized products of catechin monomers were analyzed by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry coupled with diode array detector, wherein dehydro-dimers and -trimers (oxidative coupling products of catechins' A-B ring) were found to be the major chromogenic compounds of EC and C. Additionally, the antioxidant capacity of catechin monomers only decreased after 30-min reaction, while along with further enzymatic reaction, catechin monomers presented comparable oxyradical scavenging ability (e.g., the DPPH inhibitory rates of catechin monomers were in the range of 24.42 %-50.77 %) to vitamin C (positive control, DPPH inhibitory rate was 27.66 %). Meanwhile, the inhibitory effects of most catechin monomers on α-glucosidase were enhanced in different degrees. These results provided basis for the development of enzymatically-oxidized catechin monomers as functional food color additives.

Asunto(s)

Catequina , Colorimetría , Espectrometría de Masas , Cromatografía Líquida con Espectrometría de Masas , Antioxidantes

RESUMEN

BACKGROUND: Protease 3C (3Cpro) is the only protease encoded in the human hepatitis A virus genome and is considered as a potential target for antiviral drugs due to its critical role in the viral life cycle. Additionally, 3Cpro has been identified as a potent inducer of ferroptosis, a newly described type of cell death. Therefore, studying the molecular mechanism of 3Cpro functioning can provide new insights into viral-host interaction and the biological role of ferroptosis. However, such studies require a reliable technique for producing the functionally active recombinant enzyme. OBJECTIVE: Here, we expressed different modified forms of 3Cpro with a hexahistidine tag on the N- or C-terminus to investigate the applicability of immobilized metal Ion affinity chromatography (IMAC) for producing 3Cpro. METHODS: We expressed the proteins in Escherichia coli and purified them using IMAC, followed by gel permeation chromatography. The enzymatic activity of the produced proteins was assayed using a specific chromogenic substrate. RESULTS: Our findings showed that the introduction and position of the hexahistidine tag did not affect the activity of the enzyme. However, the yield of the target protein was highest for the variant with seven C-terminal residues replaced by a hexahistidine sequence. CONCLUSION: We demonstrated the applicability of our approach for producing recombinant, enzymatically active 3Cpro.

Asunto(s)

Proteasas Virales 3C , Cromatografía de Afinidad , Escherichia coli , Histidina , Oligopéptidos , Histidina/genética , Histidina/metabolismo , Histidina/química , Proteasas Virales 3C/química , Proteasas Virales 3C/metabolismo , Humanos , Oligopéptidos/genética , Oligopéptidos/química , Oligopéptidos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Virales/genética , Proteínas Virales/química , Proteínas Virales/metabolismo , Proteínas Virales/aislamiento & purificación , Proteínas Recombinantes/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/biosíntesis , Virus de la Hepatitis A Humana/genética , Virus de la Hepatitis A Humana/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/aislamiento & purificación , Proteínas Recombinantes de Fusión/metabolismo , Proteínas Recombinantes de Fusión/biosíntesis , Cisteína Endopeptidasas/genética , Cisteína Endopeptidasas/química , Cisteína Endopeptidasas/metabolismo , Expresión Génica

RESUMEN

Organophosphorus nerve agents (OPNAs) pose a great threat to humanity. Possessing extreme toxicity, rapid lethality, and an unassuming appearance, these chemical warfare agents must be quickly and selectively identified so that treatment can be administered to those affected. Chromogenic detection is the most convenient form of OPNA detection, but current methods suffer from false positives. Here, nitrogenous base adducts of dirhodium(II,II) acetate were synthesized and used as chromogenic detectors of diethyl chlorophosphate (DCP), an OPNA simulant. UV-vis spectrophotometry was used to evaluate the sensitivity and selectivity of the complexes in the detection of DCP. Visual limits of detection (LOD) for DCP were as low as 1.5 mM DCP, while UV-vis-based LODs were as low as 0.113 µM. The dirhodium(II,II) complexes were also tested with several potential interferents, none of which produced a visual color change that could be mistaken for OPNA response. Ultimately, the Rh2(OAc)4(1,8-diazabicyclo[5.4.0]undec-7-ene)2 complex showed the best combination of detection capability and interferent resistance. These results, when taken together, show that dirhodium(II,II) paddlewheel complexes with nitrogenous base adducts can produce instant, selective, and sensitive detection of DCP. It is our aim to further explore and apply this new motif to produce even more capable OPNA sensors.

Asunto(s)

Agentes Nerviosos , Rodio , Rodio/química , Agentes Nerviosos/análisis , Agentes Nerviosos/química , Complejos de Coordinación/química , Compuestos Organofosforados/análisis , Compuestos Organofosforados/química , Límite de Detección , Compuestos Cromogénicos/química , Sustancias para la Guerra Química/análisis , Sustancias para la Guerra Química/química

RESUMEN

INTRODUCTION: Andexanet alfa (AA) - zhzo, recombinant coagulation factor Xa, is an approved antidote for oral Xa inhibitors (apixaban and rivaroxaban). Unfractionated heparin (UFH) is commonly used for therapeutic, interventional, and surgical indications. Protamine sulfate (PrSO4) is frequently used to neutralize UFH. This study aimed to investigate the comparative neutralization profiles of AA and PrSO4 for heparins of bovine, ovine, and porcine origin. MATERIALS AND METHODS: The neutralization effect of PrSO4 at 25 µg/ml and AA at 100 µg/ml was studied on an approximate surgical/interventional concentration of heparin by supplementing whole blood with each of the heparins at 25 µg/ml. For the clotting profile (activated partial thromboplastin time: aPTT), amidolytic (anti-Xa and anti-IIa), and thrombin generation assay each of the heparin were supplemented from -10-0.62 µg/ml. RESULTS: In the whole blood ACT studies, all three heparins produced strong anti-coagulant effects (400-450 seconds) compared to saline (130-150 seconds). Both AA and PrSO4 almost fully neutralized the anti-coagulant effects of heparins (140-160 seconds). Both antidotes completely reversed the anticoagulant effects of all three heparins in the aPTT and thrombin generation assay. However, PrSO4 was more effective in neutralizing the anti-Xa, and anti-IIa effects than AA, which only partially neutralized these effects. CONCLUSION: Andexanet alfa at 100 µg/ml effectively neutralizes the therapeutic and surgical/interventional concentrations of heparins in in-vitro settings. While differences in the anti-Xa, and anti-IIa effects between heparins were noted, anti-coagulant effect of these agents in the aPTT assay were comparable. A similar neutralization profile was observed in the ACT and thrombin generation assays by both agents.

Asunto(s)

Factor Xa , Antagonistas de Heparina , Heparina , Protaminas , Proteínas Recombinantes , Proteínas Recombinantes/farmacología , Factor Xa/farmacología , Bovinos , Ovinos , Porcinos , Animales , Anticoagulantes/farmacología , Heparina/farmacología , Protaminas/farmacología , Antagonistas de Heparina/farmacología , Coagulación Sanguínea/efectos de los fármacos , Tiempo de Trombina

RESUMEN

Wild birds are common hosts to numerous intracellular parasites such as single-celled eukaryotes of the family Eimeriidae (order Eucoccidiorida, phylum Apicomplexa). We investigated the infection rates, phylogeny, and pathogenicity of Isospora and Lankesterella parasites in wild and captive passerine birds. Blood and tissue samples of 815 wild and 15 deceased captive birds from Europe were tested using polymerase chain reaction and partial sequencing of the mitochondrial cytochrome b and cytochrome c oxidase I and the nuclear 18S rRNA gene. The infection rate for Lankesterella in wild birds was 10.7% compared to 5.8% for Isospora. Chromogenic in situ hybridization with probes targeting the parasites' 18S rRNA was employed to identify the parasites' presence in multiple organs, and hematoxylin-eosin staining was performed to visualize the parasite stages and assess associated lesions. Isospora parasites were mainly identified in the intestine, spleen, and liver. Extraintestinal tissue stages of Isospora were accompanied by predominantly lymphohistiocytic inflammation of varying severity. Lankesterella was most frequently detected in the spleen, lung, and brain; however, infected birds presented only a low parasite burden without associated pathological changes. These findings contribute to our understanding of Isospora and Lankesterella parasites in wild birds.

RESUMEN

Nitrofuran (NF) contamination in food products is a global problem resulting in the banned utilization and importation of nitrofuran contaminated products. A novel chromogenic detection method using a specific DNA aptamer with high affinity and specificity to nitrofurans was developed. Single-stranded DNA aptamers specific to nitrofuran metabolites, including 3-amino-2-oxazolidinone (AOZ), 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), and 1-aminohydantoin (AHD), were isolated using magnetic bead-SELEX. The colorimetric detection of nitrofurans using gold nanoparticles (AuNPs) exhibited an AOZ detection range of 0.01-0.06 ppb with a limit of detection (LOD) of 0.03 ppb. At the same time, this system could detect AMOZ and AHD at a range of 0.06 ppb and 10 ppb, respectively. The fast nitrofuran extraction method was optimized for food, such as fish tissues and honey, adjusted to be completed within 3-6 h. This novel apta-chromogenic detection method could detect NF metabolites with a sensitivity below the minimum required performance limit (MPRL). This analysis will be valuable for screening, with a shortened time of detection for aquaculture products such as shrimp and fish muscle tissues.

Asunto(s)

Aptámeros de Nucleótidos , Contaminación de Alimentos , Nanopartículas del Metal , Nitrofuranos , Nitrofuranos/análisis , Nitrofuranos/metabolismo , Nanopartículas del Metal/química , Contaminación de Alimentos/análisis , Aptámeros de Nucleótidos/química , Oxazolidinonas/análisis , Oxazolidinonas/metabolismo , Oro/química , Límite de Detección , Hidantoínas/análisis , Animales , Miel/análisis , Colorimetría/métodos , Análisis de los Alimentos/métodos

RESUMEN

BACKGROUND: The current preoperative malignancy risk evaluation for thyroid nodules involves stepwise diagnostic modalities including ultrasonography, thyroid function serology and fine-needle aspiration (FNA) cytopathology, respectively. We aimed to substantiate the stepwise contributions of each diagnostic step and additionally investigate the diagnostic significance of quantitative chromogenic imprinted gene in-situ hybridization (QCIGISH)-an adjunctive molecular test based on epigenetic imprinting alterations. METHODS: A total of 114 cytopathologically-diagnosed and histopathologically-confirmed thyroid nodules with complete ultrasonographic and serological examination records were evaluated using QCIGISH in the study. Logistic regression models for thyroid malignancy prediction were developed with the stepwise addition of each diagnostic modality and the contribution of each step evaluated in terms of discrimination performance and goodness-of-fit. RESULTS: From the baseline model using ultrasonography [area under the receiver operating characteristics curve (AUROC): 0.79; 95% confidence interval (CI): 0.71-0.86], significant improvements in thyroid malignancy discrimination were observed with the stepwise addition of thyroid function serology (AUROC: 0.82; 95% CI: 0.74-0.90; P=0.23) and FNA cytopathology (AUROC: 0.88; 95% CI: 0.81-0.94; P=0.02), respectively. The inclusion of QCIGISH as an adjunctive molecular test further advanced the preceding model's diagnostic performance (AUROC: 0.95; 95% CI: 0.91-1.00, P=0.007). CONCLUSIONS: Our study demonstrated the significant stepwise diagnostic contributions of standard clinical assessments in the malignancy risk stratification of thyroid nodules. However, the addition of molecular imprinting detection further enabled a more accurate and definitive preoperative evaluation especially for morphologically indeterminate thyroid nodules and cases with potentially discordant results among standard modalities.

Asunto(s)

Impresión Genómica , Humanos , Femenino , Masculino , Persona de Mediana Edad , Adulto , Neoplasias de la Tiroides/genética , Biopsia con Aguja Fina/métodos , Nódulo Tiroideo/genética , Anciano , Glándula Tiroides/patología

RESUMEN

The aim of this study was to assess the utility of CHROMID® Colistin R for direct detection of colistin-resistant Gram-negative bacteria from positive blood cultures. A total of 390 blood cultures from hospitalised patients containing Gram-negative bacteria were included in this study. These blood cultures were referred to clinical laboratories in the United Kingdom and Türkiye. A further 16 simulated positive blood culture bottles were included that contained a range of colistin-resistant strains as well as susceptible control strains. Fluid from each positive blood culture was diluted 1/200 in saline and 10 µL aliquots cultured onto cystine-lactose-electrolyte-deficient agar and CHROMID® Colistin R. All recovered bacteria were identified, and for Gram-negative bacteria, their minimum inhibitory concentration of colistin was measured using the broth microdilution method. From a total of 443 Gram-negative isolates, 57 colistin-resistant isolates were recovered, of which 53 (93%) grew on CHROMID® Colistin R within 18 h. Of the 377 isolates determined to be colistin-susceptible, only 9 isolates were able to grow, including 6 isolates of Pseudomonas aeruginosa. For positive blood cultures that are shown to contain Gram-negative bacteria, culture on CHROMID® Colistin R is a useful diagnostic tool to detect susceptibility or resistance to colistin within 18 h.

RESUMEN

Yeast infections are challenging human and animal medicine due to low rates of detection and the emergence of unknown ecology isolates. The aim of this study was to verify the biochemical identification of yeasts and yeast-like microorganisms obtained from animals comparing the results with chromogenic media and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF MS). Between January and August 2023, yeast and yeast-like isolates from samples of animals with suspicion of mycosis were identified using Vitek® 2 Compact, Brilliance® Candida Agar and MALDI Biotyper® MSP. A total of 39 cases were included, and 45 isolations were obtained. Cryptococcus neoformans (15.5%, 7/45), Meyerozyma guilliermondii (13.3%, 6/45), Candida parapsilosis (11.1%, 5/45), Candida albicans and Candida tropicalis (8.9%, each one 4/45) were the most identified organisms. There was full agreement with the three identification methods in 71.1% (32/45) of the isolates, disagreement on species in 17.8% (8/45), disagreement on genus and species in 6.7% (3/45) and, in 4.4% (2/45), there was no matched pattern in MALDI-TOF to compare the results. Biochemical methods are a good option in laboratories where proteomics are not available, and chromogenic media enhances diagnostics by detecting mixed infections. Surveillance must be implemented to improve the detection of agents shared between humans and animals.

RESUMEN

The design and development of a fluorescence sensor aimed at detecting and quantifying trace amounts of toxic transition metal ions within environmental, biological, and aquatic samples has garnered significant attention from diagnostic and testing laboratories, driven by the imperative to mitigate the health risks associated with these contaminants. In this context, we present the utilization of a heterocyclic symmetrical Schiff Base derivative for the purpose of fluorogenic and chromogenic detection of Co2+, Cu2+ and Hg2+ ions. The characterization of the ligand involved a comprehensive array of techniques, including physical assessments, optical analyses, NMR, FT-IR, and mass spectrometric examinations. The mechanism of ligand-metal complexation was elucidated through the utilization of photophysical parameters and FT-IR spectroscopic analysis, both before and after the interaction between the ligand and the metal salt solution. The pronounced alterations observed in absorption and fluorescence spectra, along with the distinctive chromogenic changes, following treatment with Co2+, Cu2+ and Hg2+, affirm the successful formation of complexes between the ligands and the treated metal ions. Notably, the receptor's complexation response exhibited selectivity towards Co(II), Cu(II), and Hg(II), with no observed chromogenic changes, spectral variations, or band shifts for the various tested metal ions, including Na+, Ag+, Ni2+, Mn2+, Pd2+, Pb2+, Cd2+, Zn2+, Sn2+, Fe2+, Fe3+, Cr3+ and Al3+. This absence of interaction between these metal ions and the ligand could be attributed to their compact or inadequately conducive conduction bands for complexation with the ligand's structural composition. To quantify the sensor's efficacy, fluorescence titration spectra were employed to determine the detection limits for Co2+, Cu2+ and Hg2+, yielding values of 2.92 × 10-8, 8.91 × 10-8, and 4.39 × 10-3 M, respectively. The Benesi-Hildebrand plots provided association constant values for the ligand-cobalt, ligand-copper, and ligand-mercury complexes as 0.74, 2.52, and 13.89 M-1, respectively.

RESUMEN

Detection of fluoride (F-), acetate (AcO-), and cyanide (CN-) anions is vital from the biological and environmental aspects. In the present contributions, we have introduced a simple Salen-type chromogenic sensor, BEN, to detect these biologically important anions. Changes in UV-visible absorption spectra and color of BEN solution from very pale yellow to pink color are similar for each of these anions and found to be reversible only in the case of F- ions in attendance of HSO4- ions. The estimated limit of detection of BEN solution for detecting F-, AcO-, and CN- anions is found to be below the micromolar (µM) concentration level. Our fabricated handy paper test kit is suitable for qualitatively naked-eye detection of the anions. An immediate quantitative estimation of these important anions is possible using our BEN employing a smartphone, avoiding any costly experimental setup.

RESUMEN

In 2010, the reporting of thrombotic adverse events for one subcutaneous and certain intravenous immunoglobulins (IGs) raised some concerns. In Europe, regulatory bodies rapidly revised compendial specifications for therapeutic IGs to ensure they do not exhibit thrombogenic (procoagulant) activity (PCA). At the global level, a working group (GWG) was launched with the aim of assessing PCA measurement methods and limits, considering results obtained by human IG manufacturers during in-process controls. The GWG created three dedicated subgroups to investigate the FXIa chromogenic assay, the non-activated partial thromboplastin time (NAPTT) test and the thrombin generation assay (TGA). The European Directorate for the Quality of Medicines & HealthCare (EDQM) was responsible for co-ordinating the subgroup in charge of evaluating the FXIa chromogenic assay in a study that assessed the sensitivity and robustness of two commercial chromogenic FXIa test kits. The impact of IG product formulation on FXIa recovery and the suitability of PCA-containing IG products as potential reference standards/controls were also assessed. IG materials representative of marketed products were provided to four laboratories for a study that was carried out in two steps: 1) two chromogenic FXIa test kit manufacturers assessed the performance and determined optimal test conditions by their respective methods, 2) two OMCLs studied both kits using an optimised study design. Regarding sensitivity, the study results identified suitable dose-response intervals and limits with both chromogenic FXIa test kits. This allowed the establishment of dilution ranges for optimal detection of FXIa/PCA in 5 % and 10 % IG products in the range of 1-6 mIU/mL. However, careful optimisation of the sample dilutions was required (notably to avoid potential matrix effects) and the choice of the mode of data acquisition (kinetic or end-point method) contributed to sensitivity in routine use. Importantly, the composition of IG products was of minor concern for FXIa determination with both test kits. Potential reference materials evaluated in the study behaved as expected and could be useful should a separate reference standard to the FXIa WHO IS be deemed necessary in future.

Asunto(s)

Inmunoglobulina G , Inmunoglobulinas Intravenosas , Humanos , Trombina , Pruebas de Coagulación Sanguínea , Estándares de Referencia