Fluconazole-resistant Candida parapsilosis: fast detection of the Y132F ERG11p substitution, and a proposed microsatellite genotyping scheme.

OBJECTIVES: We propose fast and accurate molecular detection of the Y132F ERG11p substitution directly on pure-cultured C. parapsilosis isolates. We also assessed a discriminative genotyping scheme to track circulating genotypes. METHODS: A total of 223 C. parapsilosis isolates (one patient each) from 20 hospitals, located in Spain and Italy were selected. Isolates were fluconazole-resistant (n=94; harbouring the Y132F ERG11p substitution [n=85], the G458S substitution [n=6], the R398I substitution [n=2], or the wild-type ERG11 gene sequence) or fluconazole-susceptible (n=129). Two targeted-A395T-mutation PCR formats (conventional and real-time) were engineered and optimized on fluconazole-susceptible and fluconazole-resistant pure-cultured isolates, thus skipping DNA extraction. Two genotyping schemes were compared: Scheme 1 (CP1, CP4a, CP6, and B markers), and Scheme 2 (6A, 6B, 6C, CP1, CP4a, and CP6 markers). RESULTS: The screening performed using both PCR formats showed 100% specificity (fluconazole-susceptible isolates; n=129/129) and sensitivity (Y132F isolates; n=85/85) values, however, results were available in 3 and 1.5 hours with the conventional and real-time PCR formats, respectively. Overall, Scheme 1 showed higher genetic diversity than Scheme 2, as shown by the number of alleles detected (n=98; mean 23, range 13-38), the significantly higher observed and expected heterozygosity, and the probability of identity index (2.5x10-6). Scheme 2 markers did not provide further genotypic discrimination of Y132F fluconazole-resistant genotypes. CONCLUSION: Both proposed PCR formats allow to speed up the accurate detection of substitution Y132F ERG11p in C. parapsilosis isolates with 100% specificity and sensitivity. In addition, we recommend CP1, CP4a, CP6, and B microsatellite markers for genotyping fluconazole-resistant isolates.

Fluconazole-resistant Candida parapsilosis genotypes from hospitals located in five Spanish cities and one in Italy: Description of azole-resistance profiles associated with the Y132F ERG11p substitution.

BACKGROUND: Fluconazole-resistant Candida parapsilosis is a matter of concern. OBJECTIVES: To describe fluconazole-resistant C. parapsilosis genotypes circulating across hospitals in Spain and Rome and to study their azole-resistance profile associated with ERG11p substitutions. PATIENTS/METHODS: We selected fluconazole-resistant C. parapsilosis isolates (n = 528 from 2019 to 2023; MIC ≥8 mg/L according to EUCAST) from patients admitted to 13 hospitals located in five Spanish cities and Rome. Additionally, we tested voriconazole, posaconazole, isavuconazole, amphotericin B, micafungin, anidulafungin and ibrexafungerp susceptibility. RESULTS: Of the 53 genotypes found, 49 harboured the Y132F substitution, five of which were dominating city-specific genotypes involving almost half the isolates. Another genotype involved isolates harbouring the G458S substitution. Finally, we found two genotypes with the wild-type ERG11 gene sequence and one with the R398I substitution. All isolates were fully susceptible/wild-type to amphotericin B, anidulafungin, micafungin and ibrexafungerp. The azole-resistance patterns found were: voriconazole-resistant (74.1%) or voriconazole-intermediate (25.2%), posaconazole-resistant (10%) and isavuconazole non-wild-type (47.5%). Fluconazole-resistant and voriconazole non-wild-type isolates were likely to harbour substitution Y132F if posaconazole was wild type; however, if posaconazole was non-wild type, substitution G458S was indicated if isavuconazole MIC was >0.125 mg/L or substitution Y132F if isavuconazole MIC was ≤0.125 mg/L. CONCLUSIONS: We detected a recent clonal spread of fluconazole-resistant C. parapsilosis across some cities in Spain, mostly driven by dominating city-specific genotypes, which involved a large number of isolates harbouring the Y132F ERG11p substitution. Isolates harbouring substitution Y132F can be suspected because they are non-susceptible to voriconazole and rarely posaconazole-resistant.

On phosphorus chemical shift anisotropy in metal (IV) phosphates and phosphonates: A 31 P NMR experimental study.

The phosphorus chemical shift anisotropies, 31 PΔcs, and asymmetry parameters η were measured by the 31 P{1 H} NMR experiments in static and low-frequency spinning samples of the zirconium phosphates and phosphonates and also in the mixed Zr (IV)/Sn (IV) phosphate/phosphonate material. The data obtained have shown a 111 connectivity in the HPO4 and PO3 groups, which does not change at modification and intercalation of the materials. The 31 PΔcs values of the phosphonate groups (43-49 ppm) significantly surpass the values characterizing the HPO4 groups (23-37 ppm). The 31 P Δcs values obtained for the metal (IV) phosphates were discussed in terms of P-O distances. The 31 P chemical shift anisotropy parameters can help at elucidation of local structures in phosphate and phosphonate materials.

Acidic Centers on the Surface of a Crystalline α-Sn(IV) Phosphate Characterized by the Solid-State 1H, 2H, 31P, and 119Sn MAS NMR Techniques.

A layered crystalline phosphate α-Sn(HPO4)2·H2O (1), prepared and characterized in the present study by the multinuclear solid-state nuclear magnetic resonance (NMR), powder X-ray diffraction, and thermogravimetric analysis techniques, was treated with D2O and HOD imitating the reaction conditions in a water medium. The 2H solid-echo magic angle spinning NMR spectra of the products have revealed on their surface low mobile water molecules and hydronium ions, forming a structure close to the Zundel cation, [D2O···D-OD2]+. All the deuterons in the hydronium ions are tangled by hydrogen bonds with the water and the surface phosphate groups and stabilized by ionic interactions.

Clinical presentation and virological assessment of confirmed human monkeypox virus cases in Spain: a prospective observational cohort study.

BACKGROUND: In May, 2022, several European countries reported autochthonous cases of monkeypox, which rapidly spread globally. Early reports suggest atypical presentations. We aimed to investigate clinical and virological characteristics of cases of human monkeypox in Spain. METHODS: This multicentre, prospective, observational cohort study was done in three sexual health clinics in Madrid and Barcelona, Spain. We enrolled all consecutive patients with laboratory-confirmed monkeypox from May 11 to June 29, 2022. Participants were offered lesion, anal, and oropharynx swabs for PCR testing. Participant data were collected by means of interviews conducted by dermatologists or specialists in sexually transmitted infections and were recorded using a standard case report form. Outcomes assessed in all participants with a confirmed diagnosis were demographics, smallpox vaccination, HIV status, exposure to someone with monkeypox, travel, mass gathering attendance, risk factors for sexually transmitted infections, sexual behaviour, signs and symptoms on first presentation, virological results at multiple body sites, co-infection with other sexually transmitted pathogens, and clinical outcomes 14 days after the initial presentation. Clinical outcomes were followed up until July 13, 2022. FINDINGS: 181 patients had a confirmed monkeypox diagnosis and were enrolled in the study. 166 (92%) identified as gay men, bisexual men, or other men who have sex with men (MSM) and 15 (8%) identified as heterosexual men or heterosexual women. Median age was 37·0 years (IQR 31·0-42·0). 32 (18%) patients reported previous smallpox vaccination, 72 (40%) were HIV-positive, eight (11%) had a CD4 cell count less than 500 cells per µL, and 31 (17%) were diagnosed with a concurrent sexually transmitted infection. Median incubation was 7·0 days (IQR 5·0-10·0). All participants presented with skin lesions; 141 (78%) participants had lesions in the anogenital region, and 78 (43%) in the oral and perioral region. 70 (39%) participants had complications requiring treatment: 45 (25%) had a proctitis, 19 (10%) had tonsillitis, 15 (8%) had penile oedema, six (3%) an abscess, and eight (4%) had an exanthem. Three (2%) patients required hospital admission. 178 (99%) of 180 swabs from skin lesions collected tested positive, as did 82 (70%) of 117 throat swabs. Viral load was higher in lesion swabs than in pharyngeal specimens (mean cycle threshold value 23 [SD 4] vs 32 [6], absolute difference 9 [95% CI 8-10]; p<0·0001). 108 (65%) of 166 MSM reported anal-receptive sex. MSM who engaged in anal-receptive sex presented with proctitis (41 [38%] of 108 vs four [7%] of 58, absolute difference 31% [95% CI 19-44]; p<0·0001) and systemic symptoms before the rash (67 [62%] vs 16 [28%], absolute difference 34% [28-62]; p<0·0001) more frequently than MSM who did not engage in anal-receptive sex. 18 (95%) of 19 participants with tonsillitis reported practising oral-receptive sex. The median time from onset of lesions to formation of a dry crust was 10 days (IQR 7-13). INTERPRETATION: In our cohort, monkeypox caused genital, perianal, and oral lesions and complications including proctitis and tonsillitis. Because of the variability of presentations, clinicians should have a low threshold for suspicion of monkeypox. Lesion swabs showed the highest viral loads, which, combined with the history of sexual exposure and the distribution of lesions, suggests close contact is probably the dominant transmission route in the current outbreak. FUNDING: None.

Elucidating structure and dynamics of crystalline α-zirconium phosphates intercalated with water and methanol by multinuclear solid-state MAS NMR: A comprehensive NMR approach.

Solid-state NMR experiments on 2 H, 31 P, 13 C, and 1 H nuclei, including 31 P T1 , 1 H T1 , and 1 H T1ρ measurements, as well as on the kinetics of proton-phosphorus cross-polarization have been performed to characterize the crystalline and amorphous α-zirconium phosphates, which were intercalated with D2 O and/or CD3 OD. The 13 C{1 H} CP MAS NMR experiment performed for compound 1-CD3 OD (Zr (HPO4 )2. 0.2CD3 OD) with carbon cross-polarization via protons of phosphate groups has provided a prove that the methanol was intercalated into the interlayer spaces of this compound. The variable-temperature 2 H solid-echo MAS NMR spectra of intercalated compounds demonstrated that the methanol molecules, in contrast to the mobile water, were immobile, keeping, however, free CD3 rotations around the C3 -axis. It has been demonstrated that the intercalated species, D2 O and CD3 OD, do not affect the high-frequency motions of the phosphate groups. By utilizing local structural models that satisfy the constraints of the experimental data, it has been suggested that the immobile methanol molecules are located in the cavity between two neighboring layers of the zirconium phosphates. Thus, the present work illustrates the reliable criteria in a comprehensive NMR approach to structural and dynamic studies of such systems.

Unexpected structural/motional mode of water intercalated into an α-crystalline zirconium phosphate deduced by 31 P and 2 H solid-state MAS NMR spectra.

In developing the approach to understanding dynamics of intercalates in layered materials, crystalline-layered zirconium phosphate Zr (HPO4 )2 ·0.35D2 O has been prepared and characterized by the 1 H, 31 P, and 2 H solid-state MAS NMR spectra, including 31 P and 2 H T1 measurements. At temperatures >253 K, the intercalated water shows two spectrally-distinguished deuterons unprecedentedly with different DQCC's and 2 H T1 times, one of which is hydrogen bonded. The collected data allowed to identify an unexpected bonding/dynamic mode of water molecules, which experience fast rotation around the hydrogen bond, formed with a zirconium-coordinated oxygen. The low-temperature 2 H MAS NMR experiments have demonstrated the presence of additional hydrogen bond P(H)O˙˙˙ DO, population of which grows on cooling to 195 K corresponding to the doubly hydrogen-bonded immobile water molecule.

SARS-CoV-2 could be spread through hospital medication dispensed to patients: A prospective observational study.

ABSTRACT: Our objective was to analyze in vitro the persistence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in the packaging material of the drugs dispensed to hospital wards. Additionally, to evaluate if the protection with a double plastic bag prevents the contamination of the medication dispensed to an intensive care unit (ICU).On the first part, different materials containing different drugs within an ICU were sampled to confirm the lack of contamination by SARS-CoV-2. The confirmation of the virus was performed using real time reverse transcription polymerase chain reaction. As a control group, in the microbiology laboratory we inoculated the virus into the different surfaces containing the same drugs included in the first part. Samples were obtained with a sterile swab at 3, 6, 8, 10, 14, 21, and 30 days after inoculation and analyzed through real time reverse transcription polymerase chain reaction.None of the studied materials containing the drugs within an ICU was contaminated by SARS-CoV-2. In the second part, SARS-CoV-2 was found in all surfaces for up to 30 days.The use of double-bag unit-dose system to deliver medication in a pandemic seems effective to prevent the potential transmission of SARS-CoV-2. A striking SARS-CoV-2 RNA stability of up to 30 days was found in the surfaces containing the drugs.

On Librational and Rotational Motions of Aromatic Rings in Layered Sn(IV) and Zr(IV) Phosphonate Materials: A Variable-Temperature 13C, 31P Solid-State NMR Study.

According to the solid-state 13C, 31P NMR study and 13C chemical shift anisotropy (CSA) measurements, aromatic rings in the layered metal(IV) phosphonate materials behave as low-energy rotors at rotation activation energy, Eact, of 1.4-3.0 kcal/mol. The rotational mechanism consists of 180° flips and librations around C(1)-C(4) axis. The amplitude of the librations, added to the flips, grows with temperature, shifting the reorientations toward rotational diffusion at high temperatures.

Solvent-Free Synthesis of Nano Zirconium Phenylphosphonates with Molten Phenylphosphonic Acid.

Nanosized α-zirconium phenylphosphonate particles were successfully prepared by the reaction between different zirconium sources and molten phenylphosphonic acid in the absence of solvent. The resultant nanoplates exhibit particle sizes in the range of 15 to 30 nm. The use of a topotactic anion exchange method starting from α-zirconium phosphate instead resulted in the generation of 15 to 180 nm plates, while also resulting in nanoparticles with a higher degree of crystallinity. The topotactic anion exchange of the phosphate groups by phenylphosphonate groups could be performed to completion when performed in molten phenylphosphonic acid. Characterization of both the final products as well as the individual steps in the anion exchange were performed by powder XRD, fast neutron activation analysis, TGA, FTIR spectroscopy, TEM, solid-state NMR and XPS.

Pyridine-d5 as a 2H NMR probe for investigation of macrostructure and pore shapes in a layered Sn(iv) phosphonate-phosphate material.

Isotropic and anisotropic motions and molecular states of pyridine-d5, adsorbed on the surface within the pores of a layered Sn(iv) phosphonate-phosphate material (1) have been characterized thermodynamically and kinetically by solid-state NMR. The data obtained provide formulation of macrostructure and shapes of pores in 1.

Zirconium Phosphate: The Pathway from Turbostratic Disorder to Crystallinity.

We show how the structural order of nanocrystalline zirconium phosphates (ZrP) is tuned by the synthetic methods and conditions through the use of synchrotron X-ray atomic pair distribution function analysis. With different synthetic route and different phosphoric acid concentrations in the synthesis, the product zirconium phosphates vary from turbostratically disordered nanoscale structures to fully ordered ones. We show that a change in the structural order leads to different ion-exchange properties. The samples are characterized using multiple techniques, including powder X-ray diffraction, ion exchange, thermogravimetric analysis, transmission electron microscopy, fast neutron activation analysis, solid-state NMR spectroscopy, IR spectroscopy, and X-ray photoelectron spectroscopy.

Comparison of 2 molecular assays and a serologic test in diagnosing Mycoplasma pneumoniae infection in paediatrics patients.

Two commercial polymerase chain reaction (PCR) assays (a real-time PCR [Cepheid] and an oligochromatographic test [Speed-oligo]) and 1 serology test (Serodia-Myco II) for detecting Mycoplasma pneumoniae in nasopharyngeal aspirates and serum samples were studied. Among the 145 samples, 32 serum pairs were serologically positive for M. pneumoniae. Of these, in 30 nasopharyngeal aspirates, M. pneumoniae was detected using the real-time PCR assay and 25 using Speed-oligo, corresponding to a sensitivity of 93.7% and 78.1%, respectively. Among the 94 samples with negative serology, we only obtained 1 positive result by real-time PCR assay. In the group of samples from healthy children, no positive results were obtained.

Mujeres en tratamiento de cáncer, acogidas por un albergue de apoyo: circunstancias y perspectivas de cuidado de enfermería / Women receiving cancer treatment supported by a housing support: circumstances and perspectives in nursing care

Se realizó un estudio de tipo cualitativo basado en 13 entrevistas a profundidad a mujeres con cáncer entre los 33 y los 70 años, con diferentes diagnósticos, provenientes de varias regiones de Colombia, vinculadas al régimen subsidiado, y que son acogidas por un albergue durante su estadía en Bogotá con motivo de su tratamiento en el Instituto Nacional de Cancerología (INC) con el fin de conocer sus circunstancias y sus necesidades de cuidado de enfermería. De las mujeres, 8 tienen un cáncer de tipo ginecológico y de este grupo 5 viven con cáncer de mama. En 4 de los 13 casos el diagnóstico fue tardío. Para todas las mujeres ha sido difícil asimilar su diagnóstico, asociándolo a la posibilidad de morir; luego ha sobrevenido una etapa de aceptación de su realidad de salud y de afrontamiento. Para este grupo, el INC y el Albergue constituyen redes sociales significativas; la lejanía de sus allegados es una dificultad que destacan del hecho de tener que desplazarse a la capital. La mayoría de las participantes buscan sentirse activas; los aspectos psicosociales son prioritarios entre sus necesidades; desean profundizar en su conocimiento de la enfermedad; es importante para ellas desarrollar mayores habilidades de auto cuidado para minimizar efectos adversos y síntomas. Fortalecer las redes de acompañamiento e interacción generadas entre ellas así como apoyarlas para fortalecer sus perspectivas de afrontamiento de su enfermedad y de autocuidado hacen parte de las estrategias que se proponen para otorgar mayor sentido a la cotidianidad y para favorecer su salud mental y su calidad de vida durante el tiempo que duran sus tratamientos en la capital.

Trauma torácico, prevención en el área laboral y conducta en las instituciones sanitarias / Thoracic trauma, prevention in the working area and behavior at the health institutions

El trauma constituye uno de los problemas más grandes de la sociedad moderna y especialmente el torácico ocupa un lugar importante con respecto a la mortalidad. Se realizó una revisión bibliográfica sobre el trauma torácico, la incidencia, evaluación y pronóstico de las lesiones, con el propósito de actualizar los conocimientos sobre su tratamiento en las instituciones sanitarias y promover medidas que disminuyan la incidencia de estos en los centros laborales(AU)

Trauma is one of the biggest problems in modern society and the thoracic trauma occupies an important place as regards mortality. A bibliographical revision about thoracic trauma, its update, incidence, evaluation and prognosis of the injuries was made in order to update the knowledge about its treatment at the health institutions and to improve measures that reduce its incidence at the working centers(AU)

Comparacion del monogliceril - formol detergente con dos tecnicas coproparasitologicas / Comparison betwen monogliceril - detergent formol with two coproparasitological techniques

Se estudiaron 2134 muestras de heces en varios laboratorios de la Red Nacional de Salud de Cuba. todas las muestras fueron observadas a dobles ciegas por las tecnicas: Directo, ritchie y Monogliceril-formol 5 por ciento (FM-5 por ciento ) con el interes de introducir una alternativa a la tecnica original de Ritchie al sustituir el reactivo eter, de importacion por el Monogliceril de produccion nacional. La tecnica MF 5 por ciento demostro tener una sensibilidad y especificidad similr a la tecnica de Ritchie y superior a la tecnica de montaje directo. Los ensayos realizados en el Instituto Pedro Kouri demostraron que el Monogliceril puede emplearse usando la centrifugacion o la sedimentacion con identicos resultados y tiene ademas valor como preservo de las heces por lo que se puede prescindir del Formol sin alterar los resultados finales. la tecnica resulta barata, sencilla y facil de realizar, por lo que es factible su extension a cualquier area de salud del pais a un costo inferior a la tecnica original de Ritchie

Control de giardiasis en una zona suburbana de la provincia de la Habana / Control of giardiasis in province ciudad de la habana

Se asignaron aleatoriamente 250 personas (49 familias) y 230 personas (52 familias) a grupos I y II respectivamente, que son objeto de dos intervenciones diferentes. Los grupos tenían en común las condiciones de saneamiento básico, eran atendidos por el mismo grupo de atención primaria y una prevalencia de G. lamblia de 21 por ciento en cada uno de ellos. Al grupo I se les administró quimioterápia con tinidazol a 50 mg/Kg de peso sin pasar de 2 gramos en dosis repetidas por 3 días y a sus miembros se les orientó con medidas de higiene que se verificaron en el hogar en un período de 30 días por dos años. Al grupo II se les indicó recetas médicas con tinidazol a igual dosis y a sus miembros se les orientó con medidas higiénicas desde la porta médica pero no se efectuaron los controles de su aplicación en el hogar. En 1992 se observó un importante descenso en la prevalencia de GH. lamblia y no se observó reinfección e incidencia en el grupo I; en cambio en el grupo II estos indicadores no se redujeron significativamente. Se recomienda que la estrategia de dispensarización del equipo médico de atención primaria unido a la quimioterapia controlada sea el método adecuado para reducir esta parasitosis en la comunidad.