RESUMEN
In a recent call to action, we described pressing issues in the health-service-psychology (HSP) internship from the perspective of interns. In our article, we sought to initiate a dialogue that would include trainees and bring about concrete changes. The commentaries on our article are a testament to the readiness of the field to engage in such a dialogue, and we applaud the actionable recommendations that they make. In our response to these commentaries, we seek to move the conversation further forward. We observe two themes that cut across these responses: the impetus to gather novel data on training (the "need to know") and the importance of taking action (the "need to act"). We emphasize that in new efforts to gather data and take policy-level action, the inclusion of trainee stakeholders (as well as others involved in and affected by HSP training) is a crucial ingredient for sustainable and equitable change.
RESUMEN
Contextual memory, the ability to remember spatial or temporal features related to an event, is affected in Alzheimer's disease (AD). There is a shortfall of tests that measure contextual memory. To evaluate visuospatial contextual memory, we developed a computerized cognitive test, the MAPP Room Memory Test, which requires participants to identify in which visual scene target items were previously presented. We hypothesized that cognitively-unimpaired carriers of an autosomal dominant AD mutation (Presenilin-1 E280A, n=15) would perform more poorly on this test than non-carrier family members (n=31). Compared to non-carriers, the carriers had significantly worse delayed room recognition. The results indicate that the MAPP Room Memory Test may be sensitive to subtle cognitive changes associated with risk of AD. Future studies with larger samples using the MAPP Room Memory Test and biomarkers are needed to examine whether this test may also be sensitive to the earliest pathological changes in preclinical AD.
Asunto(s)
Enfermedad de Alzheimer , Humanos , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/patología , Mutación , Pruebas NeuropsicológicasRESUMEN
Markerless motion capture has improved physical screening efficiency in sport and occupational settings; however, reliability of kinematic measurements from commercial systems must be established. Further, the impact of torso-borne equipment on these measurements is unclear. The purpose of this study was to evaluate the reliability of HumanTrak, a markerless motion capture system, for estimating peak trunk flexion in squat movements with and without a weighted vest. Eighteen participants completed body weight squats (BWSQ) and overhead squats (OHSQ) to their maximum depth (unrestricted-range) and to a plyometric box (fixed-range) while wearing no body armour (NBA) or 9 kg body armour (BA9). Peak trunk flexion was measured using HumanTrak. Testing was performed in two sessions on one day (intra-day) and one session on a separate day (inter-day) to assess reliability. HumanTrak had a standard error of measurement < 3.74° across all movements and conditions. Reliability was good to excellent (ICC = 0.82-0.96) with very large to nearly perfect Pearson correlations (r > 0.80) for all comparisons except unrestricted-range BWSQ with BA9 (ICC = 0.60-0.71, r = 0.71). HumanTrak was more reliable for intra- than inter-day, but reliability was still excellent for almost all inter-day comparisons (ICC > 0.82). HumanTrak is reliable for detecting differences in peak trunk flexion > 8.5° when body armour is not worn and > 10.5° when body armour is worn. Practitioners can assess meaningful changes in sagittal plane trunk motion when screening squat movements regardless of whether body armour is worn.
Asunto(s)
Captura de Movimiento , Postura , Humanos , Reproducibilidad de los Resultados , Movimiento , Rango del Movimiento Articular , Fenómenos BiomecánicosRESUMEN
Early cognitive changes due to Alzheimer's disease (AD) include difficulties in semantic access and working memory. Using a computerized cognitive test developed by our group, called the Memory for Semantically Related Objects test (MESERO), we evaluated if cognitively unimpaired carriers of an autosomal dominant AD (ADAD) mutation performed worse on this test than non-carrier family members. 35 cognitively unimpaired ADAD mutation carriers and 26 non-carrier family members from a Colombian ADAD cohort took the MESERO on a laptop computer. Cognitively unimpaired ADAD carriers had significantly worse MESERO total scores than non-carrier family members, driven by worse performance in semantically-related object sets; group performances did not differ on semantically unrelated object sets. Findings suggest that MESERO performance may be sensitive to subtle cognitive changes associated with AD. Future MESERO research should examine performances between healthy older adults and people at risk for sporadic AD.
Asunto(s)
Enfermedad de Alzheimer , Humanos , Anciano , Enfermedad de Alzheimer/psicología , Mutación/genética , Pruebas Neuropsicológicas , ColombiaRESUMEN
Susceptibility testing is an important tool in the clinical setting; its utility is based on the availability of categorical endpoints, breakpoints (BPs), or epidemiological cutoff values (ECVs/ECOFFs). CLSI and EUCAST have developed antifungal susceptibility testing, BPs, and ECVs for some fungal species. Although the concentration gradient strip bioMérieux Etest is useful for routine testing in the clinical laboratory, ECVs are not available for all agent/species; the lack of clinical data precludes development of BPs. We reevaluated and consolidated Etest data points from three previous studies and included new data. We defined ECOFFinder Etest ECVs for three sets of species-agent combinations: fluconazole, posaconazole, and voriconazole and 9 Candida spp.; amphotericin B and 3 nonprevalent Candida spp.; and caspofungin and 4 Aspergillus spp. The total of Etest MICs from 23 laboratories (Europe, the Americas, and South Africa) included (antifungal agent dependent): 17,242 Candida albicans, 244 C. dubliniensis, 5,129 C. glabrata species complex (SC), 275 C. guilliermondii (Meyerozyma guilliermondii), 1,133 C. krusei (Pichia kudriavzevii), 933 C. kefyr (Kluyveromyces marxianus), 519 C. lusitaniae (Clavispora lusitaniae), 2,947 C. parapsilosis SC, 2,214 C. tropicalis, 3,212 Aspergillus fumigatus, 232 A. flavus, 181 A. niger, and 267 A. terreus SC isolates. Triazole MICs for 66 confirmed non-wild-type (non-WT) Candida isolates were available (ERG11 point mutations). Distributions fulfilling CLSI ECV criteria were pooled, and ECOFFinder Etest ECVs were established for triazoles (9 Candida spp.), amphotericin B (3 less-prevalent Candida spp.), and caspofungin (4 Aspergillus spp.). Etest fluconazole ECVs could be good detectors of Candida non-WT isolates (59/61 non-WT, 4 of 6 species).
Asunto(s)
Anfotericina B , Candida , Anfotericina B/farmacología , Antifúngicos/farmacología , Aspergillus , Caspofungina , Pruebas Antimicrobianas de Difusión por Disco , Farmacorresistencia Fúngica , Kluyveromyces , Pruebas de Sensibilidad Microbiana , Pichia , Saccharomycetales , Triazoles/farmacologíaRESUMEN
OBJECTIVES: The primary objective of this study was to ascertain whether there is association between low initial serum progesterone, sonographic parameters and clinical outcomes in women presenting with pregnancies of unknown location (PUL), which are found to be ongoing at their follow up scans in the first trimester. STUDY DESIGN: This was a non-interventional retrospective cohort study of 1056 patients spanning a 14-year period, conducted in the Early Pregnancy Unit (EPU) of an inner-city teaching hospital. Patients who had an ongoing singleton first trimester pregnancy after presenting with PUL were identified and categorised as having low progesterone if it was 32 nmol/l or lower. The crown-rump length (CRL), mean gestational sac diameter (MGSD) and gestational sac volume (GSV) were measured when the embryo was first seen, and the pregnancy outcome recorded. RESULTS: Pregnancies with low progesterone tended to have smaller gestational sacs (GS) on follow up scan (p = 0.001) and the sac was smaller than expected for a given CRL (p = 0.000). There was no ultrasound parameter that was characteristic of low progesterone. The observation of a smaller than expected MGSD for a given CRL remained even when only pregnancies with normal outcomes were analysed. Clinical outcome data were available for 854 (80.9 %) women. Overall, 81.4 % (n = 34/43) of pregnancies with low progesterone resulted in livebirth, compared to 91.7 % (n = 744/811) livebirths in pregnancies with higher levels (p = 0.0454). CONCLUSION: Pregnancies with low progesterone tend to have a smaller GS compared to those with a higher progesterone, and the GSs are smaller than expected for a given CRL. The current study shows that women with low progesterone at the start of pregnancy remain at higher risk of miscarriage, even when the pregnancy is initially found to be viable in the first trimester. These pregnancies also tend to be associated with the sonographic finding of a smaller GS than expected for a given gestational age, regardless of eventual outcome.
Asunto(s)
Progesterona , Ultrasonografía Prenatal , Largo Cráneo-Cadera , Femenino , Edad Gestacional , Humanos , Embarazo , Primer Trimestre del Embarazo , Estudios RetrospectivosRESUMEN
INTRODUCTION: Few studies have examined how manipulating the semantic relationship between objects impacts visual working memory accuracy or reaction time. AIM: To characterize how the semantic relatedness of visual objects impacts working memory accuracy and reaction time in healthy adults using a newly developed mobile-tablet cognitive task. SUBJECTS AND METHODS: A delayed matching to sample paradigm on the tablet task was studied in a sample of 76 community-dwelling adult participants from Spain and Colombia. The tablet task included 80 unique sets of either four or six semantically related or semantically unrelated objects. The accuracy and reaction time of the participants on the task were recorded for analysis. RESULTS: When objects were semantically related, reaction time was greater in the six object sets relative to the four object sets. Age was positively associated with reaction time, but not accuracy across all four task conditions. Participants with fewer years of formal education than the sample median (16 years) exhibited worse response accuracy and slower reaction time on both the four and six semantically related conditions relative to participants with 16 or more years of formal education. CONCLUSIONS: Findings from this study suggest that when objects are semantically related (versus unrelated) and object load is increased, more processing time is needed to determine whether an object was or was not in the encoded set. The results also suggest that greater educational attainment -which likely relates with greater exposure to more technologies- is related with faster and more accurate responses on some task conditions.
TITLE: Memoria de trabajo visual para objetos relacionados semánticamente en adultos sanos.Introducción. Pocos estudios han examinado cómo la manipulación de la relación semántica entre objetos puede influir en el desempeño o el tiempo de reacción en tareas de memoria de trabajo visual. Objetivo. Caracterizar cómo la relación semántica de los objetos afecta el desempeño o el tiempo de reacción en una tarea de memoria de trabajo en adultos sanos utilizando una tarea cognitiva diseñada para el uso con tableta. Sujetos y métodos. Se usó una tarea de emparejamiento demorado (delayed matching to sample) con una muestra de 76 participantes adultos de España y Colombia. La tarea incluyó 80 conjuntos únicos de cuatro o seis objetos relacionados/no relacionados semánticamente. Se registraron las respuestas y el tiempo de reacción de los participantes. Resultados. Cuando los objetos estaban semánticamente relacionados, el tiempo de reacción fue mayor en la condición de seis objetos con respecto a la condición de cuatro objetos. La edad se asoció positivamente con el tiempo de reacción, pero no con la precisión de las respuestas. Los participantes con menos años de educación formal tuvieron un peor desempeño y un tiempo de reacción más lento en las condiciones somáticamente relacionadas en relación con los participantes con 16 años o más de educación formal. Conclusión. Los resultados sugieren que, cuando los objetos están semánticamente relacionados y aumenta su número, se necesita más tiempo de procesamiento para determinar si un objeto está o no en el grupo de objetos codificado. Además, un mayor nivel educativo se relaciona con respuestas más rápidas y un mejor desempeño en ciertas condiciones de la tarea.
Asunto(s)
Memoria a Corto Plazo , Tiempo de Reacción , Semántica , Adulto , Cognición , Colombia , Humanos , EspañaRESUMEN
Hazara nairovirus (HAZV) is an enveloped trisegmented negative-strand RNA virus classified within the Nairoviridae family of the Bunyavirales order and a member of the same subtype as Crimean-Congo hemorrhagic fever virus, responsible for fatal human disease. Nairoviral subversion of cellular trafficking pathways to permit viral entry, gene expression, assembly, and egress is poorly understood. Here, we generated a recombinant HAZV expressing enhanced green fluorescent protein and used live-cell fluorescent imaging to screen an siRNA library targeting genes involved in cellular trafficking networks, the first such screen for a nairovirus. The screen revealed prominent roles for subunits of the coat protein 1 (COPI)-vesicle coatomer, which regulates retrograde trafficking of cargo between the Golgi apparatus and the endoplasmic reticulum, as well as intra-Golgi transport. We show the requirement of COPI-coatomer subunits impacted at least two stages of the HAZV replication cycle: an early stage prior to and including gene expression and also a later stage during assembly and egress of infectious virus, with COPI-knockdown reducing titers by approximately 1,000-fold. Treatment of HAZV-infected cells with brefeldin A (BFA), an inhibitor of Arf1 activation required for COPI coatomer formation, revealed that this late COPI-dependent stage was Arf1 dependent, consistent with the established role of Arf1 in COPI vesicle formation. In contrast, the early COPI-dependent stage was Arf1 independent, with neither BFA treatment nor siRNA-mediated ARF1 knockdown affecting HAZV gene expression. HAZV exploitation of COPI components in a noncanonical Arf1-independent process suggests that COPI coatomer components may perform roles unrelated to vesicle formation, adding further complexity to our understanding of cargo-mediated transport.IMPORTANCE Nairoviruses are tick-borne enveloped RNA viruses that include several pathogens responsible for fatal disease in humans and animals. Here, we analyzed host genes involved in trafficking networks to examine their involvement in nairovirus replication. We revealed important roles for genes that express multiple components of the COPI complex, which regulates transport of Golgi apparatus-resident cargos. COPI components influenced at least two stages of the nairovirus replication cycle: an early stage prior to and including gene expression and also a later stage during assembly of infectious virus, with COPI knockdown reducing titers by approximately 1,000-fold. Importantly, while the late stage was Arf1 dependent, as expected for canonical COPI vesicle formation, the early stage was found to be Arf1 independent, suggestive of a previously unreported function of COPI unrelated to vesicle formation. Collectively, these data improve our understanding of nairovirus host-pathogen interactions and suggest a new Arf1-independent role for components of the COPI coatomer complex.
Asunto(s)
Factor 1 de Ribosilacion-ADP/genética , Factor 1 de Ribosilacion-ADP/metabolismo , Proteína Coat de Complejo I/genética , Proteína Coat de Complejo I/metabolismo , Nairovirus/genética , Nairovirus/metabolismo , Replicación Viral/fisiología , Animales , Brefeldino A , Retículo Endoplásmico/metabolismo , Regulación Viral de la Expresión Génica , Técnicas de Silenciamiento del Gen , Aparato de Golgi/metabolismo , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/fisiología , Nairovirus/patogenicidad , Transporte de Proteínas , ARN Interferente Pequeño , Replicación Viral/genéticaRESUMEN
PURPOSE: To compare the effects on ocular temperature and tear film parameters following a single application of a latent heat eyelid warming device at a range of temperature settings. METHODS: Fifteen subjects were enrolled in a prospective, investigator-masked, randomised, cross-over trial. On separate days, participants were randomised to 10-minute application of a research latent heat device (Laboratoires Théa) at device temperature settings of 45⯰C, 50⯰C and 55⯰C. Outer eyelid and corneal temperatures, tear film lipid layer grade, and non-invasive tear film breakup time (NIBUT) were measured at baseline and immediately after 10â¯min of device application. RESULTS: Baseline measurements did not differ between treatment groups (all pâ¯>â¯0.05). Ocular temperatures, lipid layer grade and non-invasive tear film stability rose significantly following device application in all treatment groups (all pâ¯<â¯0.05). The 55⯰C setting effected a mean ocular surface temperature rise in the order of +4⯰C from baseline, which was 1.46 and 1.26 times greater than at the 45⯰C and 50⯰C temperature settings, respectively (all pâ¯<â¯0.05). Similarly, improvements in mean non-invasive tear film stability from baseline in the order of +7â¯s were observed, which were 2.43 and 1.66 times greater than those at the lower temperature settings of 45⯰C and 50⯰C, respectively (all pâ¯<â¯0.05). CONCLUSIONS: At all temperature settings, the latent heat device resulted in clinically and statistically significant increases in ocular temperature, lipid layer grade, and non-invasive tear film stability. However, the 55⯰C setting proved to be most effective at raising ocular temperature (in the order of +4⯰C from baseline) and improving tear film stability.
Asunto(s)
Síndromes de Ojo Seco/terapia , Párpados/fisiopatología , Hipertermia Inducida/instrumentación , Adulto , Anciano , Anciano de 80 o más Años , Estudios Cruzados , Síndromes de Ojo Seco/fisiopatología , Diseño de Equipo , Femenino , Estudios de Seguimiento , Calor , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Resultado del Tratamiento , Adulto JovenRESUMEN
The Nairoviridae family of the Bunyavirales order comprises tick-borne, trisegmented, negative-strand RNA viruses, with several members being associated with serious or fatal diseases in humans and animals. A notable member is Crimean-Congo hemorrhagic fever virus (CCHFV), which is the most widely distributed tick-borne pathogen and is associated with devastating human disease, with case fatality rates averaging 30%. Hazara virus (HAZV) is closely related to CCHFV, sharing the same serogroup and many structural, biochemical, and cellular properties. To improve understanding of HAZV and nairovirus multiplication cycles, we developed, for the first time, a rescue system permitting efficient recovery of infectious HAZV from cDNA. This system now allows reverse genetic analysis of nairoviruses without the need for high-level biosafety containment, as is required for CCHFV. We used this system to test the importance of a DQVD caspase cleavage site exposed on the apex of the HAZV nucleocapsid protein arm domain that is cleaved during HAZV infection, for which the equivalent DEVD sequence was recently shown to be important for CCHFV growth in tick but not mammalian cells. Infectious HAZV bearing an uncleavable DQVE sequence was rescued and exhibited growth parameters equivalent to those of wild-type virus in both mammalian and tick cells, showing this site was dispensable for virus multiplication. In contrast, substitution of the DQVD motif with the similarly uncleavable AQVA sequence could not be rescued despite repeated efforts. Together, these results highlight the importance of this caspase cleavage site in the HAZV life cycle but reveal the DQVD sequence performs a critical role aside from caspase cleavage.IMPORTANCE HAZV is classified within the Nairoviridae family with CCHFV, which is one of the most lethal human pathogens in existence, requiring the highest biosafety level (BSL) containment (BSL4). In contrast, HAZV is not associated with human disease and thus can be studied using less-restrictive BSL2 protocols. Here, we report a system that is able to rescue HAZV from cDNAs, thus permitting reverse genetic interrogation of the HAZV replication cycle. We used this system to examine the role of a caspase cleavage site, DQVD, within the HAZV nucleocapsid protein that is also conserved in CCHFV. By engineering mutant viruses, we showed caspase cleavage at this site was not required for productive infection and this sequence performs a critical role in the virus life cycle aside from caspase cleavage. This system will accelerate nairovirus research due to its efficiency and utility under amenable BSL2 protocols.
Asunto(s)
Caspasas/metabolismo , Interacciones Huésped-Patógeno , Nairovirus/fisiología , Proteínas de la Nucleocápside/metabolismo , Replicación Viral , Animales , Línea Celular , ADN Complementario/genética , ADN Viral/genética , Humanos , Genética InversaRESUMEN
The Nairoviridae family within the Bunyavirales order comprise tick-borne segmented negative-sense RNA viruses that cause serious disease in a broad range of mammals, yet cause a latent and lifelong infection in tick hosts. An important member of this family is Crimean-Congo haemorrhagic fever virus (CCHFV), which is responsible for serious human disease that results in case fatality rates of up to 30â%, and which exhibits the most geographically broad distribution of any tick-borne virus. Here, we explored differences in the cellular response of both mammalian and tick cells to nairovirus infection using Hazara virus (HAZV), which is a close relative of CCHFV within the CCHFV serogroup. We show that HAZV infection of human-derived SW13 cells led to induction of apoptosis, evidenced by activation of cellular caspases 3, 7 and 9. This was followed by cleavage of the classical apoptosis marker poly ADP-ribose polymerase, as well as cellular genome fragmentation. In addition, we show that the HAZV nucleocapsid (N) protein was abundantly cleaved by caspase 3 in these mammalian cells at a conserved DQVD motif exposed at the tip of its arm domain, and that cleaved HAZV-N was subsequently packaged into nascent virions. However, in stark contrast, we show for the first time that nairovirus infection of cells of the tick vector failed to induce apoptosis, as evidenced by undetectable levels of cleaved caspases and lack of cleaved HAZV-N. Our findings reveal that nairoviruses elicit diametrically opposed cellular responses in mammalian and tick cells, which may influence the infection outcome in the respective hosts.
Asunto(s)
Apoptosis , Infecciones por Bunyaviridae/fisiopatología , Nairovirus/metabolismo , Proteínas de la Nucleocápside/metabolismo , Garrapatas/virología , Secuencias de Aminoácidos , Animales , Infecciones por Bunyaviridae/enzimología , Infecciones por Bunyaviridae/genética , Infecciones por Bunyaviridae/virología , Caspasa 3/genética , Caspasa 3/metabolismo , Caspasa 7/genética , Caspasa 7/metabolismo , Línea Celular , Interacciones Huésped-Patógeno , Humanos , Nairovirus/química , Nairovirus/genética , Proteínas de la Nucleocápside/química , Proteínas de la Nucleocápside/genética , Procesamiento Proteico-PostraduccionalRESUMEN
The study of individuals with autosomal dominant Alzheimer's disease affords one of the best opportunities to characterize the biological and cognitive changes of Alzheimer's disease that occur over the course of the preclinical and symptomatic stages. Unifying the knowledge gained from the past three decades of research in the world's largest single-mutation autosomal dominant Alzheimer's disease kindred - a family in Antioquia, Colombia with the E280A mutation in the Presenilin1 gene - will provide new directions for Alzheimer's research and a framework for generalizing the findings from this cohort to the more common sporadic form of Alzheimer's disease. As this specific mutation is virtually 100% penetrant for the development of the disease by midlife, we use a previously defined median age of onset for mild cognitive impairment for this cohort to examine the trajectory of the biological and cognitive markers of the disease as a function of the carriers' estimated years to clinical onset. Studies from this cohort suggest that structural and functional brain abnormalities - such as cortical thinning and hyperactivation in memory networks - as well as differences in biofluid and in vivo measurements of Alzheimer's-related pathological proteins distinguish Presenilin1 E280A mutation carriers from non-carriers as early as childhood, or approximately three decades before the median age of onset of clinical symptoms. We conclude our review with discussion on future directions for Alzheimer's disease research, with specific emphasis on ways to design studies that compare the generalizability of research in autosomal dominant Alzheimer's disease to the larger sporadic Alzheimer's disease population.
Asunto(s)
Enfermedad de Alzheimer/fisiopatología , Péptidos beta-Amiloides/metabolismo , Encéfalo/diagnóstico por imagen , Fragmentos de Péptidos/metabolismo , Presenilina-1/genética , Adolescente , Adulto , Enfermedad de Alzheimer/diagnóstico por imagen , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/sangre , Péptidos beta-Amiloides/líquido cefalorraquídeo , Compuestos de Anilina , Enfermedades Asintomáticas , Biomarcadores/sangre , Biomarcadores/líquido cefalorraquídeo , Biomarcadores/metabolismo , Encéfalo/metabolismo , Encéfalo/fisiopatología , Niño , Colombia , Imagen de Difusión Tensora , Progresión de la Enfermedad , Electroencefalografía , Glicoles de Etileno , Femenino , Neuroimagen Funcional , Humanos , Imagen por Resonancia Magnética , Masculino , Pruebas de Estado Mental y Demencia , Persona de Mediana Edad , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/líquido cefalorraquídeo , Tomografía de Emisión de Positrones , Radiofármacos , Tomografía Computarizada de Emisión de Fotón Único , Adulto JovenRESUMEN
Hyperforin (HYP), one of the main bioactive compounds in extracts of Hypericum perforatum, is a potential drug candidate for the treatment of skin diseases. Since extracts have proven to support wound healing, in the present study effects of HYP on human dermal fibroblasts (HDF) were evaluated in 2D and 3D in vitro dermal constructs. Viability and cytotoxicity assays as well as a live-dead cell staining were performed to test at which concentration HYP reduces viability and/or shows cytotoxicity. Furthermore a differentiation between cytotoxic, anti-proliferative and anti-migratory effects was done. For the latter purpose a 2D migration assay was performed. HDF-induced contraction of a 3D artificial dermal (AD) construct was determined at given HYP concentration. Induction of apoptosis was examined by determination of caspase 3/7 activities. HYP reduced viability of HDF down to 70% at concentrations of 5-10µM. This decrease was not due to cytotoxicity but to a reduction in proliferation as shown from both the proliferation assay and the cytotoxicity assay as well as from live-dead cell staining. The 2D migration assay showed that HYP reduced migration activity of HDF cells at a concentration of 10µM. At this concentration HYP also reduced the HDF-induced contraction of collagen gels as 3D AD constructs. Apoptotic effects of HYP were excluded performing a caspase 3/7 activity detecting assay. The results show for the first time that HYP may be rather a potential candidate for treatment of hypertrophic scars than promoting effects which are understood as important in wound healing.
Asunto(s)
Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Cicatriz Hipertrófica , Fibroblastos/efectos de los fármacos , Prepucio/efectos de los fármacos , Floroglucinol/análogos & derivados , Terpenos/farmacología , Animales , Movimiento Celular/fisiología , Proliferación Celular/fisiología , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Células Cultivadas , Cicatriz Hipertrófica/tratamiento farmacológico , Cicatriz Hipertrófica/patología , Relación Dosis-Respuesta a Droga , Fibroblastos/fisiología , Prepucio/citología , Prepucio/fisiología , Humanos , Recién Nacido , Masculino , Floroglucinol/farmacología , Floroglucinol/uso terapéutico , Ratas , Terpenos/uso terapéutico , Resultado del Tratamiento , Cicatrización de Heridas/efectos de los fármacos , Cicatrización de Heridas/fisiologíaRESUMEN
Due to the limited chemical stability of the natural hyperforin molecule, a more stable form of hyperforin, i.e., the hyperforin dicyclohexylammonium salt (HYP-DCHA) has been used for ex vivo and in vitro experiments in recent years, but its actual stability under typical cell culture conditions has never been studied before. In this contribution the stability of HYP-DCHA was examined under typical cell culture conditions. Different cell culture media with and without fetal calf serum (FCS) supplementation were studied with regard to further stabilization of HYP-DCHA determined with HPLC analysis. Furthermore, albumin nanoparticles were examined as a stabilizing carrier system for HYP-DCHA. In this context, the interaction between HYP-DCHA and albumin nanoparticles (ANP) was examined with regard to size and loading with HYP . The effects of HYP-DCHA either supplied in cell culture medium or loaded on ANP on viability and cytotoxicity were studied in vitro on HaCaT monolayers (human keratinocyte cell line). HYP-DCHA supplied in FCS-containing medium was recovered completely after 24h of incubation. However, a lack of FCS caused a total loss of HYP-DCHA after less than 24h incubation time. Supplying HYP-DCHA loaded on ANP in an FCS-free medium resulted in a recovery of about 60% after 24h incubation. HYP-DCHA supplied in medium along with FCS showed a slow dose-dependent decrease in viability of HaCaT cells without any cytotoxic effects (antiproliferative effect). Treatment with HYP-DCHA with a lack of FCS resulted in a significantly faster decrease in viability which was mainly due to cytotoxicity. The latter was true for HYP-DHCA-loaded ANP where increased cytotoxicity was observed despite the presence of FCS. The results show that the stability of the widely used HYP-DCHA is rather limited under cell culture conditions. Especially a lack of FCS leads to degradation and/or oxidation of HYP-DCHA probably causing an increased cytotoxicity. In contrast, FCS supplementation fairly stabilizes HYP-DCHA under cell culture conditions while albumin nanoparticles may serve the same stabilization purpose despite increasing cytotoxic effects onto the cells themselves.
Asunto(s)
Queratinocitos/efectos de los fármacos , Nanopartículas/química , Albúmina Sérica Bovina/química , Albúmina Sérica Bovina/farmacología , Terpenos/química , Terpenos/farmacología , Técnicas de Cultivo de Célula/métodos , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Estabilidad de Medicamentos , Humanos , Queratinocitos/fisiología , Nanopartículas/administración & dosificación , Espectrometría de Masa por Ionización de Electrospray/métodosRESUMEN
An indicator of movement quality and potential injury risk during Functional Movement Screen (FMS) testing is the presence of asymmetry when comparing the left and right sides of the body. The aim of the study was to investigate the reproducibility of the injury risk model proposed in our previous research (Chalmers et al. 2017; derivation study) that showed an increased injury risk for elite junior Australian football players demonstrating ≥2 asymmetrical FMS subtests. We used a direct replication design. Players underwent pre-season FMS testing, and an injury surveillance system monitored 277 male participants during the subsequent regular season competition. Designated club officials monitored the weekly competition participation of players. The definition of an injury was "a trauma or medical condition which caused a player to miss a competitive game". Cox proportional hazards regression models were used to investigate the relationship between asymmetry and number of games played before first injury (ie, survival time). The level of reproducibility was determined according to statistical significance, effect size, and subjective assessment. Demonstrating asymmetry during FMS testing was not associated with a significant increase in prospective injury risk in the replication study (P > .05). Moreover, effect sizes (hazard ratios) from the derivation dataset were not within the 95% confidence intervals of the respective asymmetry predictor in the replication dataset. Subjectively, researchers were in agreement that the findings from the derivation data were not successfully reproduced. Clinicians and researchers should be cautious about using FMS asymmetry findings to derive injury risk for junior football players.
Asunto(s)
Traumatismos en Atletas/diagnóstico , Movimiento , Fútbol/lesiones , Adolescente , Australia , Humanos , Masculino , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Reproducibilidad de los Resultados , Factores de RiesgoRESUMEN
AIMS: To assess the independent role of severe hypoglycemia on 7-year cumulative incidence of prolonged QTc in a large cohort of patients with type 1 diabetes. METHODS: People with type 1 diabetes recruited by the EURODIAB Prospective Complications Study who had normal QTc were examined at baseline and after 7 years with standardized methods (n = 1415; mean age ± SD 32.1 ± 9.6 years; diabetes duration 14.2 ± 8.8 years). Hypoglycemic episodes were assessed by a questionnaire. QTc was calculated according to Bazett's formula. In logistic regression analysis, we examined the role of severe hypoglycemia (none, 1-2, or 3 and more episodes/year) on the cumulative incidence of prolonged QTc, independently of age, sex, HbA1c, blood pressure, BMI, physical activity, distal symmetrical and autonomic neuropathy. RESULTS: In total, 264/1415 (17%) patients had incident prolonged QTc. Compared to those with persistently normal QTc, a greater proportion of incident cases had 3 and more hypoglycemic episodes at baseline (16.3 vs 11.2%, p = 0.03) and after 7 years (15.2 vs 9.6%, p = 0.01). In logistic regression analysis, 3 or more episodes of severe hypoglycemia at baseline did not increase cumulative incidence of prolonged QTc (OR 1.34, 95% CI 0.88-2.03). By contrast, severe hypoglycemia at the follow-up examination was associated with higher incidence of QTc prolongation (OR 1.68, 1.09-2.58), which reverted to not significant after adjustment for diabetic neuropathy. CONCLUSIONS: Severe hypoglycemia was not associated with incidence QTc prolongation in type 1 diabetic patients from the EURODIAB PCS.
Asunto(s)
Diabetes Mellitus Tipo 1/epidemiología , Angiopatías Diabéticas/epidemiología , Hipoglucemia/epidemiología , Síndrome de QT Prolongado/epidemiología , Adulto , Presión Sanguínea , Estudios de Cohortes , Diabetes Mellitus Tipo 1/complicaciones , Angiopatías Diabéticas/complicaciones , Neuropatías Diabéticas/complicaciones , Neuropatías Diabéticas/epidemiología , Femenino , Estudios de Seguimiento , Humanos , Hipoglucemia/etiología , Incidencia , Síndrome de QT Prolongado/complicaciones , Masculino , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
The CLSI epidemiological cutoff values (ECVs) of antifungal agents are available for various Candida spp., Aspergillus spp., and the Mucorales. However, those categorical endpoints have not been established for Fusarium spp., mostly due to the difficulties associated with collecting sufficient CLSI MICs for clinical isolates identified according to the currently recommended molecular DNA-PCR-based identification methodologies. CLSI MIC distributions were established for 53 Fusarium dimerum species complex (SC), 10 F. fujikuroi, 82 F. proliferatum, 20 F. incarnatum-F. equiseti SC, 226 F. oxysporum SC, 608 F. solani SC, and 151 F. verticillioides isolates originating in 17 laboratories (in Argentina, Australia, Brazil, Canada, Europe, Mexico, and the United States). According to the CLSI guidelines for ECV setting, ECVs encompassing ≥97.5% of pooled statistically modeled MIC distributions were as follows: for amphotericin B, 4 µg/ml (F. verticillioides) and 8 µg/ml (F. oxysporum SC and F. solani SC); for posaconazole, 2 µg/ml (F. verticillioides), 8 µg/ml (F. oxysporum SC), and 32 µg/ml (F. solani SC); for voriconazole, 4 µg/ml (F. verticillioides), 16 µg/ml (F. oxysporum SC), and 32 µg/ml (F. solani SC); and for itraconazole, 32 µg/ml (F. oxysporum SC and F. solani SC). Insufficient data precluded ECV definition for the other species. Although these ECVs could aid in detecting non-wild-type isolates with reduced susceptibility to the agents evaluated, the relationship between molecular mechanisms of resistance (gene mutations) and MICs still needs to be investigated for Fusarium spp.
Asunto(s)
Antifúngicos/farmacología , Fusarium/efectos de los fármacos , Pruebas de Sensibilidad Microbiana/métodos , Américas , Farmacorresistencia Fúngica Múltiple , Europa (Continente) , Fusarium/genética , Fusarium/aislamiento & purificación , Humanos , Reacción en Cadena de la Polimerasa/métodosRESUMEN
BACKGROUND/OBJECTIVES: Diet and lifestyle advice for type 1 diabetes (T1DM) patients is based on little evidence and putative effects on glycaemic control. Therefore, we investigated the longitudinal relation between dietary and lifestyle variables and HbA1c levels in patients with type 1 diabetes. SUBJECTS/METHODS: A 7-year prospective cohort analysis was performed in 1659 T1DM patients (52% males, mean age 32.5 years) participating in the EURODIAB Prospective Complications Study. Baseline dietary intake was assessed by 3- day records and physical activity, smoking status and alcohol intake by questionnaires. HbA1c during follow-up was centrally assessed by immunoassay. Analysis of variance (ANOVA) and restricted cubic spline regression analyses were performed to assess dose-response associations between diet and lifestyle variables and HbA1c levels, adjusted for age, sex, lifestyle and body composition measures, baseline HbA1c, medication use and severe hypoglycaemic attacks. RESULTS: Mean follow-up of our study population was 6.8 (s.d. 0.6) years. Mean HbA1c level was 8.25% (s.d. 1.85) (or 66.6 mmol/mol) at baseline and 8.27% (s.d. 1.44) at follow-up. Physical activity, smoking status and alcohol intake were not associated with HbA1c at follow-up in multivariable ANOVA models. Baseline intake below the median of vegetable protein (<29 g/day) and dietary fibre (<18 g/day) was associated with higher HbA1c levels. Restricted cubic splines showed nonlinear associations with HbA1c levels for vegetable protein (P (nonlinear)=0.008) and total dietary fibre (P (nonlinear)=0.0009). CONCLUSIONS: This study suggests that low intake of vegetable protein and dietary fibre are associated with worse glycaemic control in type 1 diabetes.
Asunto(s)
Diabetes Mellitus Tipo 1/sangre , Dieta/efectos adversos , Conducta Alimentaria , Hemoglobina Glucada/análisis , Adolescente , Adulto , Análisis de Varianza , Glucemia/metabolismo , Registros de Dieta , Fibras de la Dieta/efectos adversos , Proteínas en la Dieta/efectos adversos , Ejercicio Físico , Femenino , Estudios de Seguimiento , Humanos , Estilo de Vida , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Análisis de Regresión , Fumar , Encuestas y Cuestionarios , Verduras , Adulto JovenRESUMEN
Primary localized amyloidosis of the airways is an uncommon disorder characterized by amyloid deposits in the airway mucosa. In contrast to systemic amyloidosis other organs are not involved. Among the entities of airway amyloidosis, tracheobronchial amyloidosis is comparatively the most common subtype in the lower respiratory tract and laryngeal amyloidosis in the upper respiratory tract. The pathophysiology of localized airway amyloidosis is poorly understood. The clinical presentation is variable and often non-specific. No general consensus exists with regard to optimal treatment resulting in a variety of modalities used in clinical practice to manage this disorder. We report the case of a 50 year old woman with multifocal localized amyloidosis of the tracheobronchial tree and the upper airways. Tracheobronchial amyloidosis was treated with endoscopic debulking and external beam radiation, sinunasal amyloid deposits were surgically excised and are currently under surveillance. The importance of this extremely rare case lies in the multifocal presentation of an uncommon disorder requiring a multidisciplinary approach to offer optimal treatment including external beam radiation.