Neuropsychological and Anatomical-Functional Effects of Transcranial Magnetic Stimulation in Post-Stroke Patients with Cognitive Impairment and Aphasia: A Systematic Review.

Transcranial magnetic stimulation (TMS) has been found to be promising in the neurorehabilitation of post-stroke patients. Aphasia and cognitive impairment (CI) are prevalent post-stroke; however, there is still a lack of consensus about the characteristics of interventions based on TMS and its neuropsychological and anatomical-functional benefits. Therefore, studies that contribute to creating TMS protocols for these neurological conditions are necessary. To analyze the evidence of the neuropsychological and anatomical-functional TMS effects in post-stroke patients with CI and aphasia and determine the characteristics of the most used TMS in research practice. The present study followed the PRISMA guidelines and included articles from PubMed, Scopus, Web of Science, ScienceDirect, and EMBASE databases, published between January 2010 and March 2023. In the 15 articles reviewed, it was found that attention, memory, executive function, language comprehension, naming, and verbal fluency (semantic and phonological) are the neuropsychological domains that improved post-TMS. Moreover, TMS in aphasia and post-stroke CI contribute to greater frontal activation (in the inferior frontal gyrus, pars triangularis, and opercularis). Temporoparietal effects were also found. The observed effects occur when TMS is implemented in repetitive modality, at a frequency of 1 Hz, in sessions of 30 min, and that last more than 2 weeks in duration. The use of TMS contributes to the neurorehabilitation process in post-stroke patients with CI and aphasia. However, it is still necessary to standardize future intervention protocols based on accurate TMS characteristics.

Real-life application of a stratification model for HIV care.

INTRODUCTION: HIV infection has become a chronic disease with a good long-term prognosis, necessitating a change in the care model. For this study, we applied a proposal for an Optimal Care Model (OCM) for people with HIV (PHIV), which includes tools for assessing patient complexity and their classification into profiles to optimize care provision. METHODS: Observational, cross-sectional, and retrospective study. Adult PHIV treated at the Tropical Medicine consultations at Ramón y Cajal Hospital from January 1 to June 30, 2023, were included. The complexity calculation and the stratification into profiles for each patient were done according to the OCM. RESULTS: Ninety-four participants were included, 76.6% cisgender men, with a median age of 41 years (range 23-76). Latin America and Africa were the main regions of origin (72.4%). 98% had an undetectable HIV viral load. The degree of complexity was 78.7% low, 11.7% medium, 1% high, and 8.5% extreme. The predominant profile was blue (64.9%), followed by lilac (11.7%), purple (6.3%), and green (4.3%). 7.4% were unclassifiable, of whom 57.2% had high/extreme complexity. Among the unclassifiable, mental health problems were the most common. CONCLUSIONS: The OCM tools for People Living with HIV (PLWH) allow for the classification and stratification of most patients in a consultation with a non-standard population. Patients who did not fit into the pre-established profiles presented high complexity. Creating a profile focused on mental health or mixed profiles could facilitate the classification of more patients.

Changes in Calprotectin (S100A8-A9) and Aldolase in the Saliva of Horses with Equine Gastric Ulcer Syndrome.

Equine gastric ulcer syndrome (EGUS) is a highly prevalent disease that affects horses worldwide. Within EGUS, two different forms have been described: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). The associated clinical signs cause detrimental activity performance, reducing the quality of life of animals. Saliva can contain biomarkers for EGUS that could be potentially used as a complementary tool for diagnosis. The objective of this work was to evaluate the measurements of calprotectin (CALP) and aldolase in the saliva of horses as potential biomarkers of EGUS. For this purpose, automated assays for the quantification of these two proteins were analytically validated and applied for detecting EGUS in a total of 131 horses divided into 5 groups: healthy horses, ESGD, EGGD, combined ESGD and EGGD, and horses with other intestinal pathologies. The assays showed good precision and accuracy in analytical validation, and they were able to discriminate between horses with EGUS and healthy horses, especially in the case of CALP, although they did not show significant differences between horses with EGUS and horses with other diseases. In conclusion, salivary CALP and aldolase can be determined in the saliva of horses and further studies are warranted to elucidate the potential of these analytes as biomarkers in EGUS.

Evaluation of a Comprehensive Profile of Salivary Analytes for the Diagnosis of the Equine Gastric Ulcer Syndrome.

In this report, the measurement of salivary biomarkers as an aid for diagnosis of equine gastric ulcer syndrome (EGUS) was studied. A comprehensive panel of 23 salivary analytes was measured in the saliva of horses affected by EGUS and compared to healthy animals and horses with other diseases clinically similar to EGUS but with a negative diagnosis at gastroscopic examination. A total of 147 horses were included in the study and divided into heathy population (n = 12), the EGUS group (n = 110), and the group of horses with other diseases (n = 25). From the 23 analytes studied, 17 showed increased values in EGUS horses when compared to healthy ones, and uric acid, triglycerides, and calcium were significantly increased in horses with EGUS compared to the group of other diseases. The receiver operating characteristic curve analyses showed a modest but significant discriminatory power of those three analytes to identify EGUS from other diseases with similar symptoms. The discriminatory power enhanced when the results of the three analytes were combined. In conclusion, the results showed that selected salivary analytes could have potential use as biomarkers in horses with EGUS.

Role of the Triplet State and Protein Dynamics in the Formation and Stability of the Tryptophan Radical in an Apoazurin Mutant.

The protein, azurin, has enabled the study of the tryptophan radical. Upon UV excitation of tyrosine-deficient apoazurin and in the presence of a Co(III) electron acceptor, the neutral radical (W48•) is formed. The lifetime of W48• in apoazurin is 41 s, which is shorter than the lifetime of several hours in Zn-substituted azurin. Molecular dynamics simulations revealed enhanced fluctuations of apoazurin which likely destabilize W48•. The photophysics of W48 was investigated to probe the precursor state for ET. The phosphorescence intensity was eliminated in the presence of an electron acceptor while the fluorescence was unchanged; this quenching of the phosphorescence is attributed to ET. The kinetics associated with W48• were examined with a model that incorporates intersystem crossing, ET, deprotonation, and decay of the cation radical. The estimated rate constants for ET (6 × 106 s-1) and deprotonation (3 × 105 s-1) are in agreement with a photoinduced mechanism where W48• is derived from the triplet state. The triplet as the precursor state for ET was supported by photolysis of apoazurin with 280 nm in the absence and presence of triplet-absorbing 405 nm light. Absorption bands from the neutral radical were observed only in the presence of blue light.

Bimolecular quenching of tryptophan fluorescence in a membrane protein: Evolution of local solvation and environment during folding into a bilayer.

Fluorescence spectroscopy, including Stern-Volmer quenching, is a valuable tool for the study of protein dynamics. Changes in protein solvation during the folding reaction of a membrane protein, Outer membrane protein A (OmpA), into lipid bilayers was probed with bimolecular fluorescence quenching with acrylamide quencher. Six single-tryptophan OmpA mutants (W7, W15, W57, W102, W129, and W143) allowed for site-specific investigations at varying locations within the transmembrane ß-barrel domain. A sphere-of-action quenching model that combines both static and dynamic components gave rise to Stern-Volmer quenching constants, KD, for OmpA denatured in 8.0 M urea, aggregated in 0.5 M urea, adsorbed onto small unilamellar vesicles (SUVs), and folded in SUVs (t = 6 hrs). The average KD values were KDdenatured(6.4M-1)>KDaggregated5.9M-1>KDadsorbed(1.9M-1)>KDfolded(0.6M-1). With knowledge of the fluorescence lifetimes in the absence of quencher, the bimolecular quenching constants, kq, were derived; the evolution of kq (and therefore KD)during the folding reaction into SUVs (t = 0 hr to t = 6 hrs) revealed desolvation timescales, τdesolv of 41-46 min (W7, W15, W57, W102), 27 min (W129), and 15 min (W143). The evolution of λmax during folding revealed fast and slow components, τenvironmentfast and τenvironmentslow of 7-13 min and 25-84 min, respectively, for all mutants. For the five lipid- facing mutants (W7, W15, W57, W129, and W143), the general trend was τenvironmentfast7-13min<τdesolv15-46min≤τenvironmentslow(25-84min). These results suggest that there is an initial fast step in which there is a large change in polarity to a hydrophobic environment, followed by a slower desolvation process during evolution within the hydrophobic environment. These results complement previous mechanisms of concerted folding and provide insights into site-specific changes in solvation during formation of native ß-barrel structure.

Functional results of swallowing and aspiration after oral cancer treatment and microvascular free flap reconstruction: A retrospective observational assessment.

This study aimed to assess the functional swallowing outcomes in cohort of oral cancer patients treated with tumor resection and reconstruction by means of microvascular free flaps. Duration from onset to the last examination was classified into three periods: less than 12 months, from 13 to 24 months, and more than 24 months. Type of feeding, dysphagia, and laryngeal aspiration were the dependent variables, and the study was mainly focused on the data from the multivariate analysis. Fifty-one patients were included in the study. Twenty-one patients had dysphagia, 11 showed stasis in vallecula and pyriform sinuses, 9 experienced laryngeal penetration, and 8 aspirations. Surgery combined with radiochemotherapy was associated with a higher prevalence of dysphagia (p = 0.03). Frequency of dysphagia was higher in the first year after treatment and less frequent after 24 months (p = 0.003). Dysphagia was associated with laryngeal penetration (p = 0.001), and this with aspiration (p < 0.0005). In conclusion, as it seems that the method of reconstruction has a major influence on swallowing, when there are relevant alternatives in the way the reconstruction is done, always the approach should be chosen that reduces dysphagia to a minimum.

A Silent Corticotroph Pituitary Carcinoma: Lessons From an Exceptional Case Report.

Nowadays, neither imaging nor pathology evaluation can accurately predict the aggressiveness or treatment resistance of pituitary tumors at diagnosis. However, histological examination can provide useful information that might alert clinicians about the nature of pituitary tumors. Here, we describe our experience with a silent corticothoph tumor with unusual pathology, aggressive local invasion and metastatic dissemination during follow-up. We present a 61-year-old man with third cranial nerve palsy at presentation due to invasive pituitary tumor. Subtotal surgical approach was performed with a diagnosis of silent corticotroph tumor but with unusual histological features (nuclear atypia, frequent multinucleation and mitotic figures, and Ki-67 labeling index up to 70%). After a rapid regrowth, a second surgical intervention achieved successful debulking. Temozolomide treatment followed by stereotactic fractionated radiotherapy associated with temozolomide successfully managed the primary tumor. However, sacral metastasis showed up 6 months after radiotherapy treatment. Due to aggressive distant behavior, a carboplatine-etoposide scheme was decided but the patient died of urinary sepsis 31 months after the first symptoms. Our case report shows how the presentation of a pituitary tumor with aggressive features should raise a suspicion of malignancy and the need of follow up by multidisciplinary team with experience in its management. Metastases may occur even if the primary tumor is well controlled.

Early perfusion changes in multiple sclerosis patients as assessed by MRI using arterial spin labeling.

BACKGROUND: Gadolinium-perfusion magnetic resonance (MR) identifies gray matter abnormalities in early multiple sclerosis (MS), even in the absence of structural differences. These perfusion changes could be related to the cognitive disability of these patients, especially in the working memory. Arterial spin labeling (ASL) is a relatively recent perfusion technique that does not require intravenous contrast, making the technique especially attractive for clinical research. PURPOSE: To verify the perfusion alterations in early MS, even in the absence of cerebral volume changes. To introduce the ASL sequence as a suitable non-invasive method in the monitoring of these patients. MATERIAL AND METHODS: Nineteen healthy controls and 28 patients were included. The neuropsychological test EDSS and SDMT were evaluated. Cerebral blood flow and bolus arrival time were collected from the ASL study. Cerebral volume and cortical thickness were obtained from the volumetric T1 sequence. Spearman's correlation analyzed the correlation between EDSS and SDMT tests and perfusion data. Differences were considered significant at a level of P < 0.05. RESULTS: Reduction of the cerebral blood flow and an increase in the bolus arrival time were found in patients compared to controls. A negative correlation between EDSS and thalamus transit time, and between EDSS and cerebral blood flow in the frontal cortex, was found. CONCLUSION: ASL perfusion might detect changes in MS patients even in absent structural volumetric changes. More longitudinal studies are needed, but perfusion parameters could be biomarkers for monitoring these patients.

Phosphorylated ribosomal protein S6 correlation with p21 expression and inverse association with tumor size in oral squamous cell carcinoma.

BACKGROUND: The purpose of this study was to investigate the clinical relevance of phosphorylated ribosomal protein S6 (p-S6), a surrogate marker of mammalian target of rapamycin (mTOR) activation, and p21 in a series of 125 patients with oral squamous cell carcinomas (OSCCs). METHODS: Immunohistochemical analysis was performed to ascertain the phosphorylation status of p-S6 at Ser235/236 and Ser240/244, p21, and p53 protein expression. RESULTS: Expression of phosphorylated S6 protein on either serine 235/236 or serine 240/244 was detected in 83% and 88% tumors, respectively, and both of them were inversely and significantly correlated with the tumor size and local infiltration. Positive p21 expression was found in 91.5% of the cases, and was inversely correlated with tumor size. In OSCC, p21 expression correlates with p-S6 levels, a surrogate marker of mTOR activation, independently of p53 status. CONCLUSION: Expression of both p21 and p-S6 was found to inversely associate with tumor size but not survival outcomes in patients with OSCC.

Insights into Protein Structure and Dynamics by Ultraviolet and Visible Resonance Raman Spectroscopy.

Raman spectroscopy is a form of vibrational spectroscopy based on inelastic scattering of light. In resonance Raman spectroscopy, the wavelength of the incident light falls within an absorption band of a chromophore, and this overlap of excitation and absorption energy greatly enhances the Raman scattering efficiency of the absorbing species. The ability to probe vibrational spectra of select chromophores within a complex mixture of molecules makes resonance Raman spectroscopy an excellent tool for studies of biomolecules. In this Current Topic, we discuss the type of molecular insights obtained from steady-state and time-resolved resonance Raman studies of a prototypical photoactive protein, rhodopsin. We also review recent efforts in ultraviolet resonance Raman investigations of soluble and membrane-associated biomolecules, including integral membrane proteins and antimicrobial peptides. These examples illustrate that resonance Raman is a sensitive, selective, and practical method for studying the structures of biological molecules, and the molecular bonding, geometry, and environments of protein cofactors, the backbone, and side chains.

Relevance of level IIb neck dissection in oral squamous cell carcinoma.

BACKGROUND: The purpose of this study was to determine the prevalence of level IIb metastasis in patients with oral squamous cell carcinomas (OSCCs). MATERIAL AND METHODS: A prospective analysis of 56 patients with OSCC who underwent surgical treatment of the primary lesion with simultaneous neck dissection was performed. During neck dissection, level IIb lymph nodes were separately removed and processed. Neck dissection was bilateral in 26 patients (46%) and unilateral in 30 patients (54%). RESULTS: The mean number of nodes found in the level IIb specimens was 4.7 (range: 0-8 nodes). The prevalence of metastasis at level IIb was 0% in pN0 necks and 3.4% in pN+ necks, with an overall prevalence of 1.8%. A significant association between metastasis to level IIb and type of neck dissection was observed. There were no isolated metastases to level IIb without the involvement of other nodes in the remaining neck specimen. Four regional recurrences were observed during follow-up. CONCLUSIONS: Based on our findings, we suggest that dissection of the level IIb region in patients with OSCC may be required only in patients with multilevel neck metastasis or if level IIa metastasis is found intraoperatively.

Tissue-engineered oral mucosa for mucosal reconstruction in a pediatric patient with hemifacial microsomia and ankyloglossia.

Many types of soft tissue grafts have been used for the reconstruction of oral mucosal defects. The best results are achieved with mucosal grafts; however, when large areas must be grafted, sufficient donor tissue is not available. Tissue engineering represents an alternative method to obtain sufficient autologous tissue for reconstructing oral wounds. Herein we present a pediatric patient with hemifacial microsomia and congenital ankyloglossia requiring multiple surgical interventions, and in which an autologous full-thickness tissue-engineered oral mucosa was used for successful oral reconstruction. Our study demonstrates that even under challenging conditions, robust tissue-engineered products, such as the fibrin-based oral mucosa described here, can achieve successful tissue regeneration.

Epidemiología de la mortalidad infantil en la provincia Granma / Infant Mortality Epidemiology in Granma Province

Introducción: la mortalidad infantil refleja la panorámica socioeconómica de un país, define su nivel de vida influido por factores ecológicos y de atención médica. Objetivos: caracterizar epidemiológicamente la mortalidad infantil de la provincia Granma durante los años 2001 a 2011, así como, pronosticar su comportamiento en el 2012. Métodos: se emplearon técnicas de análisis de las siguientes series cronológicas de mortalidad infantil: tasas, causas de muerte, componente neonatal (precoz y tardía), posneonatal y fallecidos por meses. Se utilizaron frecuencias absolutas, relativas y tasas; los resultados se mostraron básicamente a través del gráfico aritmético simple. Se empleó el método de alisamiento exponencial para analizar tendencia, método de Holt para pronosticar la tasa, y el multiplicativo para la estacionalidad. Resultados: la mortalidad infantil mostró tendencia cíclica descendente, para este 2012 se pronosticó tasa entre 3,6 y 5,4. Las principales causas de muerte fueron las afecciones perinatales (49,4%), malformaciones congénitas (30,9%), infecciones respiratorias agudas (10,2%) y sepsis (9,5%), que mostraron un comportamiento muy inestable. Predominó el componente posneonatal con tendencia descendente en los últimos cinco años y la mortalidad infantil mostró patrón estacional. Conclusiones: la mortalidad infantil en la provincia presenta tendencia descendente similar al país aunque con picos que revelan resultados inconsistentes.

Introduction: infant mortality reflects a country's socioeconomic panoramic that defines the standard of living influenced by environmental factors and health care. Objective: to characterize the epidemiology of infant mortality in Granma province from 2001 to 2011 as well as to predict its behaviour in 2012. Methods: the following techniques of analysis of infant mortality were used: rates, causes of death, neonatal component (early and late), and post-neonatal deaths per month were used. Absolute frequencies, relative and rates were used. The results were basically through simple arithmetic graph. Exponential smoothing method was used to analyze trend and the Holt method was used to predict the rate, and multiplicative seasonality. Results: infant mortality showed cyclical trend downward for 2012 is forecast between 3.6 and 5.4 rate. The main causes of death were perinatal conditions (49.4%), congenital malformations (30.9%), acute respiratory infections (10.2%) and sepsis (9.5%), which showed very unstable behavior. Postneonatal component predominated with downward trend in the last five years and infant mortality showed a seasonal pattern. Conclusions: infant mortality in the province has similar downward trend in the country but with peaks reveal inconsistent results.

In oral squamous cell carcinoma, high FAK expression is correlated with low P53 expression.

Focal adhesion kinase (FAK) and p53 have been associated with metastatic activity and a poor prognosis in oral squamous cell carcinoma (OSCC). Recently, a feedback mechanism in which FAK regulates p53 has been proposed. The present study aims to determine the role of p53 in FAK regulation in these tumors. FAK and p53 expression was examined by immunohistochemistry in normal oral mucosa and in 67 oral squamous cell carcinomas. p16(INK4a) was also studied in view of its association with human papillomavirus infection. The association between FAK and p53 was subsequently analyzed. FAK expression in OSCCs was heterogeneous: 22 (33 %) cases showed weak expression, 16 (24 %) showed moderate expression, and 22 (33 %) cases showed high expression. Regarding p53, 31 of 67 (46 %) available tumor specimens showed negative staining, and 36 of 67 (54 %) showed positive nuclear staining for p53. FAK expression was inversely correlated with p53 expression (Fisher's exact test, p = 0.005). There was no association between p16(INK4a) and p53 or FAK expression. In conclusion, our results support the hypothesis that FAK activity might be involved in the down-regulation of p53 expression in OSCC.

Clinical outcomes after the use of complete autologous oral mucosa equivalents: preliminary cases.

OBJECTIVE: Previously, we reported how to obtain complete autologous oral mucosa equivalents (CAOMEs) composed of an autologous plasma scaffold and fibroblasts together with immature keratinocytes able to build an oral epithelium with a structure similar to that of the oral mucosa. In this study, we present the clinical outcomes after applying our CAOMEs as grafts. STUDY DESIGN: Four patients who needed a CAOME to restore a defect of oral mucosa were selected. Two of the patients suffered from ankyloglossia, and the other 2 required a restoration of the keratinized gum of the alveolar rim. To assess the outcomes, the scale designed by Ewers et al. was used. RESULTS: Clinical and functional improvements were achieved in the patients with ankyloglossia. In cases of gum restoration, the mucosa was regenerated and a prosthetic restoration with implants was achieved. CONCLUSIONS: The results obtained points to the potential use of CAOME in intraoral lining.

Use of autologous skin equivalents with artificial dermal matrix (integra) in donor site coverage in radial forearm free flaps: preliminary cases.

PURPOSE: The radial forearm flap is one of the most commonly used methods for intraoral reconstruction in oral carcinoma surgery. One of its disadvantages is the residual functional and unaesthetic defect in the donor site. The objective of this report is to describe preliminary cases of a novel method to cover such donor sites based on the use of autologous skin equivalents (ASEs) and an artificial dermal matrix (Integra, Prim, Barcelona, Spain). MATERIALS AND METHODS: The donor sites of 2 patients were treated with the artificial dermal matrix after raising a radial forearm flap. A skin biopsy and a blood sample were taken to construct an ASE. After 3 weeks, the ASE was applied over the dermal template and left to heal. The functional and esthetic results were recorded. RESULTS: Good functional and esthetic results were achieved, with correct wrist motility, although a natural skin color could not be achieved. Neither the Integra nor the ASE was rejected. Total wound coverage was achieved at 4 months, and completely normal skin was observed at 6 months. CONCLUSIONS: This technique of combining an artificial dermal matrix with an ASE could be an alternative method to cover the donor sites of radial forearm flaps.

Förster resonance energy transfer as a probe of membrane protein folding.

The folding reaction of a ß-barrel membrane protein, outer membrane protein A (OmpA), is probed with Förster resonance energy transfer (FRET) experiments. Four mutants of OmpA were generated in which the donor fluorophore, tryptophan, and acceptor molecule, a naphthalene derivative, are placed in various locations on the protein to report the evolution of distances across the bilayer and across the protein pore during a folding event. Analysis of the FRET efficiencies reveals three timescales for tertiary structure changes associated with insertion and folding into a synthetic bilayer. A narrow pore forms during the initial stage of insertion, followed by bilayer traversal. Finally, a long-time component is attributed to equilibration and relaxation, and may involve global changes such as pore expansion and strand extension. These results augment the existing models that describe concerted insertion and folding events, and highlight the ability of FRET to provide insight into the complex mechanisms of membrane protein folding. This article is part of a Special Issue entitled: Membrane protein structure and function.