Molecular histopathology of matrix proteins through autofluorescence super-resolution microscopy.

Extracellular matrix diseases like fibrosis are elusive to diagnose early on, to avoid complete loss of organ function or even cancer progression, making early diagnosis crucial. Imaging the matrix densities of proteins like collagen in fixed tissue sections with suitable stains and labels is a standard for diagnosis and staging. However, fine changes in matrix density are difficult to realize by conventional histological staining and microscopy as the matrix fibrils are finer than the resolving capacity of these microscopes. The dyes further blur the outline of the matrix and add a background that bottlenecks high-precision early diagnosis of matrix diseases. Here we demonstrate the multiple signal classification method-MUSICAL-otherwise a computational super-resolution microscopy technique to precisely estimate matrix density in fixed tissue sections using fibril autofluorescence with image stacks acquired on a conventional epifluorescence microscope. We validated the diagnostic and staging performance of the method in extracted collagen fibrils, mouse skin during repair, and pre-cancers in human oral mucosa. The method enables early high-precision label-free diagnosis of matrix-associated fibrotic diseases without needing additional infrastructure or rigorous clinical training.

Scalable-resolution structured illumination microscopy.

Structured illumination microscopy suffers from the need of sophisticated instrumentation and precise calibration. This makes structured illumination microscopes costly and skill-dependent. We present a novel approach to realize super-resolution structured illumination microscopy using an alignment non-critical illumination system and a reconstruction algorithm that does not need illumination information. The optical system is designed to encode higher order frequency components of the specimen by projecting PSF-modulated binary patterns for illuminating the sample plane, which do not have clean Fourier peaks conventionally used in structured illumination microscopy. These patterns fold high frequency content of sample into the measurements in an obfuscated manner, which are de-obfuscated using multiple signal classification algorithm. This algorithm eliminates the need of clean peaks in illumination and the knowledge of illumination patterns, which makes instrumentation simple and flexible for use with a variety of microscope objective lenses. We present a variety of experimental results on beads and cell samples to demonstrate resolution enhancement by a factor of 2.6 to 3.4 times, which is better than the enhancement supported by the conventional linear structure illumination microscopy where the same objective lens is used for structured illumination as well as collection of light. We show that the same system can be used in SIM configuration with different collection objective lenses without any careful re-calibration or realignment, thereby supporting a range of resolutions with the same system.

Colonización nasal por Staphylococcus aureus y Staphylococcus aureusresistente a meticilina (SARM) en auxiliares de enfermería / Nasal carriage of Staphylococcus aureus and Methicillin-resistant Staphylococcusaureus (MRSA) among nursing assistants

Introducción: Staphylococcus aureus, es la principal causa de bacteriemia infecciosa y endocarditis, así como de infecciones osteoarti-culares, de piel y tejidos blandos, su reservorio principal es la mucosa nasal. Los trabajadores de la salud son una fuente importante de transmisión de S. aureus y S. aureus resistente a la meticilina (SARM). Objetivo: determinar la presencia de Staphylococcus aureus y SARM en la fosa nasal de auxiliares de enfermería en la ciudad de Bogotá. Materiales y métodos: estudio descriptivo de corte transversal, en auxiliares de enfermería de diferentes instituciones hospitalarias y clínicas en la ciudad de Bogotá, Colombia. Se realizó un muestreo aleatorio. El tamaño de la muestra fue de 491 hisopados de la fosa nasal derecha de igual número de auxiliares de enfermería que al momento del estudio se encontraban laborando a nivel clínico. Se tomó un intervalo de confianza del 95% y error máximo admisible del 5%, se consideró el valor de p= 0,5. Se realizó un estudio de frecuencias y determinación de prevalencias mediante un análisis univariado. Resultados: la presente investigación encontró queel 28,5% de los participantes fueron portadores del Staphylococcus aureus y el 6,1% fueron SARM. Conclusiones: la colonización por Staphylococcus aureus y SARM es frecuente en auxiliares de enfermería.

Introduction: Staphylococcus aureus is a human pathogen of clinical severe relevance, it is the leading cause of infectious bacteremia and endocarditis, as well as osteoarticular, skin, and soft tissue infections; its, main reservoir is the nasal mucosa. Healthcare workers are a significant source of transmission of S. aureus and methicillin-resistant S. aureus (MRSA). Objective: To determine the presence of Staphylococcus aureus and MRSA in the nostrils of nursing assistants in Bogotá's city. Materials and methods: a descriptive cross-sec-tional study in nursing assistants from different hospitals and clinical institutions in Bogotá's city, Colombia. Random sampling was carried out. The sample size was 491 swabs from the right nostril from the same number of nursing assistants working at the clinical level at the time of the study. A confidence interval of 95% and maximum permissible error of 5% were taken, the value of p = 0.5 was considered. A study of frequencies and determination of prevalence was carried out through univariate analysis. Results: the present investigation found that 28.5% of the participants were carriers of Staphylococcus aureus, and 6.1% were methicillin-resistant S. aureus(MRSA). Conclusions: colonization by Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) is common in nursing assistants.

Chip-based multimodal super-resolution microscopy for histological investigations of cryopreserved tissue sections.

Histology involves the observation of structural features in tissues using a microscope. While diffraction-limited optical microscopes are commonly used in histological investigations, their resolving capabilities are insufficient to visualize details at subcellular level. Although a novel set of super-resolution optical microscopy techniques can fulfill the resolution demands in such cases, the system complexity, high operating cost, lack of multi-modality, and low-throughput imaging of these methods limit their wide adoption for histological analysis. In this study, we introduce the photonic chip as a feasible high-throughput microscopy platform for super-resolution imaging of histological samples. Using cryopreserved ultrathin tissue sections of human placenta, mouse kidney, pig heart, and zebrafish eye retina prepared by the Tokuyasu method, we demonstrate diverse imaging capabilities of the photonic chip including total internal reflection fluorescence microscopy, intensity fluctuation-based optical nanoscopy, single-molecule localization microscopy, and correlative light-electron microscopy. Our results validate the photonic chip as a feasible imaging platform for tissue sections and pave the way for the adoption of super-resolution high-throughput multimodal analysis of cryopreserved tissue samples both in research and clinical settings.

Fluorescence fluctuation-based super-resolution microscopy using multimodal waveguided illumination.

Photonic chip-based total internal reflection fluorescence microscopy (c-TIRFM) is an emerging technology enabling a large TIRF excitation area decoupled from the detection objective. Additionally, due to the inherent multimodal nature of wide waveguides, it is a convenient platform for introducing temporal fluctuations in the illumination pattern. The fluorescence fluctuation-based nanoscopy technique multiple signal classification algorithm (MUSICAL) does not assume stochastic independence of the emitter emission and can therefore exploit fluctuations arising from other sources, as such multimodal illumination patterns. In this work, we demonstrate and verify the utilization of fluctuations in the illumination for super-resolution imaging using MUSICAL on actin in salmon keratocytes. The resolution improvement was measured to be 2.2-3.6-fold compared to the corresponding conventional images.

Deriving high contrast fluorescence microscopy images through low contrast noisy image stacks.

Contrast in fluorescence microscopy images allows for the differentiation between different structures by their difference in intensities. However, factors such as point-spread function and noise may reduce it, affecting its interpretability. We identified that fluctuation of emitters in a stack of images can be exploited to achieve increased contrast when compared to the average and Richardson-Lucy deconvolution. We tested our methods on four increasingly challenging samples including tissue, in which case results were comparable to the ones obtained by structured illumination microscopy in terms of contrast.

Artefact removal in ground truth deficient fluctuations-based nanoscopy images using deep learning.

Image denoising or artefact removal using deep learning is possible in the availability of supervised training dataset acquired in real experiments or synthesized using known noise models. Neither of the conditions can be fulfilled for nanoscopy (super-resolution optical microscopy) images that are generated from microscopy videos through statistical analysis techniques. Due to several physical constraints, a supervised dataset cannot be measured. Further, the non-linear spatio-temporal mixing of data and valuable statistics of fluctuations from fluorescent molecules that compete with noise statistics. Therefore, noise or artefact models in nanoscopy images cannot be explicitly learned. Here, we propose a robust and versatile simulation-supervised training approach of deep learning auto-encoder architectures for the highly challenging nanoscopy images of sub-cellular structures inside biological samples. We show the proof of concept for one nanoscopy method and investigate the scope of generalizability across structures, and nanoscopy algorithms not included during simulation-supervised training. We also investigate a variety of loss functions and learning models and discuss the limitation of existing performance metrics for nanoscopy images. We generate valuable insights for this highly challenging and unsolved problem in nanoscopy, and set the foundation for the application of deep learning problems in nanoscopy for life sciences.

Soft thresholding schemes for multiple signal classification algorithm.

Multiple signal classification algorithm (MUSICAL) exploits temporal fluctuations in fluorescence intensity to perform super-resolution microscopy by computing the value of a super-resolving indicator function across a fine sample grid. A key step in the algorithm is the separation of the measurements into signal and noise subspaces, based on a single user-specified parameter called the threshold. The resulting image is strongly sensitive to this parameter and the subjectivity arising from multiple practical factors makes it difficult to determine the right rule of selection. We address this issue by proposing soft thresholding schemes derived from a new generalized framework for indicator function design. We show that the new schemes significantly alleviate the subjectivity and sensitivity of hard thresholding while retaining the super-resolution ability. We also evaluate the trade-off between resolution and contrast and the out-of-focus light rejection using the various indicator functions. Through this, we create significant new insights into the use and further optimization of MUSICAL for a wide range of practical scenarios.

Periodontally Accelerated Osteogenic Orthodontics (PAOO) technique in cleft patients: A complement to orthognathic surgery in dentoalveolar expansion. A case series report.

The aim of this study was to describe a surgical technique that can be used to solve dentofacial deformities in cleft palate patients with maxillary hypoplasia in order to increase maxillary alveolar bone width, without modifying the skeletal base, and therefore, keeping the velopharyngeal function unaltered. Four patients with a history of cleft palate not associated with syndrome and treated under conventional surgical protocol during their childhood, underwent PAOO surgery incorporating L-PRF, followed by an accelerated orthodontic treatment with checkups every two weeks. All patients reached the desired occlusion without modifying their skeletal bases and velopharyngeal function. Orthodontic treatments were finished between 10 and 14 months after surgery without complications. There were no observed complications in the velopharyngeal postsurgical function and an increase in the arch width was achieved in all cases, along with a reduced orthodontic treatment time. The clinical results obtained confirm that PAOO technique is a safe and reliable complement to orthognathic surgery in the surgical treatment of cleft patients. By increasing the perimeter of the maxillary dentoalveolar ridge, the segmentation of the maxilla could be avoided, increasing the post operatory stability of these patients.

MusiJ: an ImageJ plugin for video nanoscopy.

We present an open-source implementation of the fluctuation-based nanoscopy method MUSICAL for ImageJ. This implementation improves the algorithm's computational efficiency and takes advantage of multi-threading to provide orders of magnitude faster reconstructions than the original MATLAB implementation. In addition, the plugin is capable of generating super-resolution videos from large stacks of time-lapse images via an interleaved reconstruction, thus enabling easy-to-use multi-color super-resolution imaging of dynamic systems.

Cáncer Bucomaxilofacial en Niños III: Tumores Malignos del Tejido Blando / Oral and Maxillofacial Cancer in Children III: Malignant Soft Tissue Tumors

RESUMEN: El grupo de neoplasias malignas de tejido blando de la región de cabeza y cuello en pacientes pediátricos está representado por carcinomas, sarcomas, melanomas y tumores de diferenciación incierta. La neoplasia más prevalente en la población pediátrica es el Rabdomiosarcoma, seguido por el carcinoma de células escamosas. Los rangos de presentación son muy amplios, siendo los grupos entre 2-6 años y 15-19 años los que presentan mayor incidencia. Se ha planteado que la etiología de estas neoplasias es incierta. El tratamiento de estas neoplasias es comúnmente de enfoque multimodal, combinando un procedimiento quirúrgico con quimioterapia y radioterapia. El pronóstico y sobrevida del paciente dependerán principalmente del momento en que se realice el diagnóstico de la lesión. Un diagnóstico y tratamiento temprano favorecen las posibilidades de sobrevida y el pronóstico del paciente. Este estudio corresponde a la 3ra parte de "Cáncer bucomaxilofacial en niños". Se hará referencia a los distintos tumores malignos del tejido blando en la población pediátrica en el territorio de cabeza y cuello, abarcando sus generalidades, etiología, epidemiología, tratamiento y pronóstico.

ABSTRACT: Head and neck malignant tumors in pediatric patients comprise carcinoma, sarcoma, melanoma and tumours of uncertain differentiation. Within the pediatric population, the most prevalent neoplasm is rhabdomyosarcoma, followed by squamous cell carcinoma. There is a wide range in the presentation, and it varies significantly with age groups of 2-6 and 15-19 year-olds who present the higher incidence rates. For this reason, it has been suggested that the etiology of head and neck neoplasms remains unclear. Treating these pathologies usually involves a multimodal approach that combines surgery, radiation and chemotherapy. Prognosis and survival rates depend mainly of the stage at the time of diagnosis. Early diagnosis and treatment can improve prognosis and survival rates. In this 3rd part of "Maxillofacial Cancer in Pediatric Patients", we studied a variety of malignant tumors in head and neck soft tissue from a paediatric sample. Specifically, we aim to analyze their etiology, epidemiology, treatment and prognosis.

Calidad del agua y características habitacionales de un barrio en Bogotá / Water quality and housing characteristics of a neighborhood in Bogota

Resumen Introducción. El agua suministrada a las viviendas debe ser apta para el consumo humano, es decir, debe ser limpia, no tener color ni sabor y sobre todo debe estar libre de microbios y parásitos. Objetivo. Se indagó la calidad microbiológica y fisicoquímica del agua de un barrio marginal de Bogotá; asimismo se determinaron las condiciones habitacionales. Método. Estudio descriptivo transversal. Se realizó un muestreo del agua utilizada para el consumo humano en 25 casas, a la cual se le realizó el análisis fisicoquímico y microbiológico. Además se diligenció un formato de condiciones de vivienda. Conclusiones. se evidenció que la calidad de agua que consumen los habitantes del barrio Villa Cindy cumple con todos los parámetros de potabilidad exigidos el Ministerio de Salud y Protección Social, de Ambiente y Desarrollo Territorial en la Resolución 2115 de 2007. El 80% de los habitantes del barrio viven en condiciones dignas, un 20% son invasores de la ribera del río Bogotá y habitan en condiciones indignas.

Abstract Introduction. The water supplied to households must be suitable for human consumption, it means, should be clean, have no color or taste, but above all must be free of microbes and parasites. Objective. Microbiological and physicochemical water properties were determined in a Bogotá slum; also, the living conditions were determined. Method. Descriptive cross-sectional study. Water used for human consumption were sampled in 25 houses, which underwent the physicochemical and microbiological analysis. In addition, a format of housing conditions was filled out. Conclusions. It was observed that the quality of water consumed by the residents of the neighborhood Villa Cindy meets all potability parameters required by Ministry of Health and Social Protection, Environment and Territorial Development in Resolution 2115 of 2007. 80% of the local people was living in decent conditions, 20% were invading the hillside of the Bogota river and lived in inhuman conditions.