Validation and implementation of telecytology at an academic medical center using digital cameras and Microsoft Teams software.

INTRODUCTION: Rapid On-Site Evaluation of cytological samples obtained through fine needle aspiration for adequacy is a critical component of a cytology service; however, it imposes a significant time and cost burden for the practicing pathologist and the cytology service. Telecytology enables adequacy assessment by a pathologist remotely, greatly saving time. Telecytology also allows slide preparation and manipulation at the procedure site by an employee with less training requirements, liberating the cytotechnologist to screen cases and perform other laboratory duties - an important aspect to consider during times of cytotechnologist shortages. We propose a telecytology system with a simple setup of a microscope, microscope camera, laptop, and Microsoft Teams software. MATERIALS AND METHODS: We designed a system consisting of a mobile cart, backup battery, microscope, digital camera, and a laptop computer with microscope imaging software and Microsoft Teams software for image transmission. Validation was performed by 4 pathologists making adequacy assessments on randomly selected previously signed out cases using the telecytology system. RESULTS: Our validation of this system demonstrated a greater than 90% concurrence rate between the original adequacy call and the call made by pathologists using the telecytology system - a benchmark used by most, if not all, published validations of similar telecytology systems. In addition, the adequacy assessment concordance rate between select pathologists exceeded 90%. CONCLUSIONS: In summary, our telecytology system provides excellent adequacy services for the clinicians and patients we serve. The Microsoft Teams software is a great tool for transmission of video microscopy. This system will be used with the goal of saving time and increasing efficiency for the cytopathology department.

Anti-GD2 CAR-NKT cells in relapsed or refractory neuroblastoma: updated phase 1 trial interim results.

Vα24-invariant natural killer T cells (NKTs) have anti-tumor properties that can be enhanced by chimeric antigen receptors (CARs). Here we report updated interim results from the first-in-human phase 1 evaluation of autologous NKTs co-expressing a GD2-specific CAR with interleukin 15 (IL15) (GD2-CAR.15) in 12 children with neuroblastoma (NB). The primary objectives were safety and determination of maximum tolerated dose (MTD). The anti-tumor activity of GD2-CAR.15 NKTs was assessed as a secondary objective. Immune response evaluation was an additional objective. No dose-limiting toxicities occurred; one patient experienced grade 2 cytokine release syndrome that was resolved by tocilizumab. The MTD was not reached. The objective response rate was 25% (3/12), including two partial responses and one complete response. The frequency of CD62L+NKTs in products correlated with CAR-NKT expansion in patients and was higher in responders (n = 5; objective response or stable disease with reduction in tumor burden) than non-responders (n = 7). BTG1 (BTG anti-proliferation factor 1) expression was upregulated in peripheral GD2-CAR.15 NKTs and is a key driver of hyporesponsiveness in exhausted NKT and T cells. GD2-CAR.15 NKTs with BTG1 knockdown eliminated metastatic NB in a mouse model. We conclude that GD2-CAR.15 NKTs are safe and can mediate objective responses in patients with NB. Additionally, their anti-tumor activity may be enhanced by targeting BTG1. ClinicalTrials.gov registration: NCT03294954 .

Epidemiological Characterization of Patients in the First Eight Weeks Following Detection of SARS-CoV-2 B.1.1.529 (omicron) Variant in Cuba.

INTRODUCTION: In November 2021, omicron-a new SARS-CoV-2 variant-was identified in South Africa and almost immediately, WHO declared it a 'variant of concern'. In view of its rapid worldwide spread and its imminent introduction in Cuba, genomic surveillance was strengthened. OBJECTIVE: Describe cases during the first eight epidemiological weeks (epiweeks) of SARS-CoV-2 infection attributable to omicron variant in Cuba by clinical and epidemiological variables. METHODS: From epiweek 48, 2021 to epiweek 4, 2022, 288 nasopharyngeal swabs were processed for sequencing of a 1836 bp fragment of the S gene. Variants were identified according to GISAID database and confirmed by phylogenetic analysis. Variants' association with clinical and epidemiological outcomes was assessed. RESULTS: The first cases of omicron variant were imported, mostly from African countries and the United States. During the period studied, omicron was detected in 83.0% (239/288) of cases processed, while the delta variant was found in 17.0% (49/288). Most persons infected with omicron were symptomatic (63.2%; 151/239) and fully vaccinated (65.3%; 156/239); severe cases and deaths occurred mainly among patients aged ≥65 years (92.9%; 13/14), and 12 of these deaths occurred in fully vaccinated persons (92.3%; 12/13). Omicron spread rapidly throughout the country (from 10% of cases in epiweek 48, 2021, to 100% by epiweek 4, 2022), displacing the formerly predominant delta variant. CONCLUSIONS: Omicron's rapid expansion in Cuba was associated with increased incidence but not with a higher case fatality rate. The relatively milder disease in those infected with this variant could be influenced by the high vaccination coverage, along with the natural immunity acquired as a consequence of previous virus infection.

Characterizing genes associated with cancer using the CRISPR/Cas9 system: A systematic review of genes and methodological approaches.

The CRISPR/Cas9 system enables a versatile set of genomes editing and genetic-based disease modeling tools due to its high specificity, efficiency, and accessible design and implementation. In cancer, the CRISPR/Cas9 system has been used to characterize genes and explore different mechanisms implicated in tumorigenesis. Different experimental strategies have been proposed in recent years, showing dependency on various intrinsic factors such as cancer type, gene function, mutation type, and technical approaches such as cell line, Cas9 expression, and transfection options. However, the successful methodological approaches, genes, and other experimental factors have not been analyzed. We, therefore, initially considered more than 1,300 research articles related to CRISPR/Cas9 in cancer to finally examine more than 400 full-text research publications. We summarize findings regarding target genes, RNA guide designs, cloning, Cas9 delivery systems, cell enrichment, and experimental validations. This analysis provides valuable information and guidance for future cancer gene validation experiments.

Metastatic colorectal micropapillary carcinoma presenting as lymphangitic lung carcinomatosis.

Pulmonary lymphangitic carcinomatosis denotes the infiltration of tumor cells into the lung parenchymal lymphatic channels. Breast, lung, stomach, and colon adenocarcinoma are the most common origin of this invasion pattern. The micropapillary variant of colorectal adenocarcinoma has a high rate of lymph node metastases and poor overall survival. A 49 year-old man with a 6 months history of persistent cough and a relevant occupational chemical exposure had a computed tomography that showed bilateral interstitial lung infiltrates. The lung biopsy demonstrated a micropapillary adenocarcinoma with diffusely obstruction of the lung parenchymal lymphatics. The immunohistochemistry confirmed a colorectal origin. The colonoscopy evidenced a mass with identical morphology. Colorectal micropapillary carcinoma with metastatic lung lymphangitic carcinomatosis can occur, as a persistent cough, as presenting symptom in extraordinarily rare cases. To the best of our knowledge, this is the first case of an alive patient with colorectal metastatic micropapillary carcinoma presenting with lymphangitic lung carcinomatosis.

Transcriptomic and cellular analyses of CRISPR/Cas9-mediated edition of FASN show inhibition of aggressive characteristics in breast cancer cells.

Several genes are significantly mutated in breast cancer but only a small percentage of mutations are well-known to contribute to cancer development. FASN is involved in de novo lipogenesis and the regulation of ERα signaling. However, the effect of genetic mutations affecting FASN in breast cancer has not thoroughly studied. Therefore, we used the CRISPR/Cas9 system to edit the FASN locus in MCF-7 cells and evaluated its biological effect. We obtained four clones carrying mutations and frameshifts in the acyl-transferase domain of FASN. We found that clones had reduced proliferation, migration, viability, and showed alterations in cell cycle profiles. RNA-Seq analysis demonstrates that a lack of fully functional FASN may have a more significant role in proliferation-related genes than in lipid metabolism. We conclude that functional knockouts in FASN contributes to decrease the proliferation and migration of breast cancer cells contrary to point mutations in breast cancer patients.

What a city eats: Examining the dietary preferences of families living in communities at high risk for food insecurity.

INTRODUCTION: Food insecurity (FI) is the lack of consistent access to enough food for an active and healthy life. Community-based hunger relief programs often serve as emergency food sources for families with FI. However, these programs may not provide foods that diverse populations of people prefer. We sought to evaluate the dietary patterns and preferences of families living in food-insecure neighborhoods and utilizing a community-based hunger relief program, in order to improve the utilization of local nutritional programs. METHODS: We examined the Help Our People Eat (HOPE) community-based mobile meal program. Free-listing interviews (n = 63) were conducted with English-(66%) and Spanish-speaking (34%) participants of the program. Participants were asked about FI risk, food preferences, and dietary behaviors at home. RESULTS: The majority of participants (90%) had children in the household. About 60% reported not being able to afford the type of food they enjoyed. Most participants reported using stoves for cooking (80%). Participants overwhelmingly cooked with chicken, beef, and pork. The most common side dishes included potatoes, rice, and salad. Most participants reported no interest in cooking differently or learning new recipes. CONCLUSIONS: A common theme throughout interviews was that families prefer similar meals, but may prepare them differently based on the language spoken. Food preferences consisted of a high intake of carbohydrate-rich meals, perhaps because these foods may be cheaper and easier to access. Notably, new recipes and cooking methods were not a priority for these families, possibly due to the time and effort needed to learn them.

Temporal and age-dependent effects of haptoglobin deletion on intracerebral hemorrhage-induced brain damage and neurobehavioral outcomes.

Intracerebral hemorrhage (ICH) is a devastating stroke subtype and the presence of extracorpuscular hemoglobin (Hb) exacerbates brain damage. Haptoglobin (Hp) binds Hb, which prevents its oxidation and participation in neurotoxic reactions. Multiple studies have investigated the role of Hp under conditions of intravascular hemolysis, but little is known about its role in the brain and following ICH where extravascular hemolysis is rampant. Young and aged wildtype and Hp-/- mice underwent the autologous blood or collagenase ICH model. Early after ICH, Hp-/- mice display 58.0 ±â€¯5.6% and 36.7 ±â€¯6.9% less brain damage in the autologous blood and collagenase ICH models, respectively. In line with these findings, Hp-/- mice display less neurological deficits on several neurobehavioral tests. Hp-/- mice have less Perl's iron content, HO1 expression, and blood brain barrier dysfunction, but no difference in brain Hb content, astrogliosis and angiogenesis/neovascularization. At the later endpoint, the young cohort displays 27.8 ±â€¯9.3% less brain damage, while no difference is seen with the aged cohort. For both cohorts, no differences are seen in HO1 levels or iron accumulation, but young Hp-/- mice display less thalamic astrogliosis and striatal microgliosis. This study reveals that the presence or absence of Hp exerts important time- and age-dependent influences on ICH outcomes.

The absence of the CD163 receptor has distinct temporal influences on intracerebral hemorrhage outcomes.

Hemoglobin (Hb) toxicity precipitates secondary brain damage following intracerebral hemorrhage (ICH). CD163 is an anti-inflammatory Hb scavenger receptor and CD163-positive macrophages/microglia locally accumulate post-bleed, yet no studies have investigated the role of CD163 after ICH. ICH was induced in wildtype and CD163-/- mice and various anatomical and functional outcomes were assessed. At 3 d, CD163-/- mice have 43.4 ± 5.0% (p = 0.0002) and 34.8 ± 3.4% (p = 0.0003) less hematoma volume and tissue injury, respectively. Whereas, at 10 d, CD163-/- mice have 49.2 ± 15.0% larger lesions (p = 0.0385). An inflection point was identified, where CD163-/- mice perform better on neurobehavioral testing and have less mortality before 4 d, but increased mortality and worse function after 4 d (p = 0.0389). At 3 d, CD163-/- mice have less Hb, iron, and blood-brain barrier dysfunction, increased astrogliosis and neovascularization, and no change in heme oxygenase 1 (HO1) expression. At 10 d, CD163-/- mice have increased iron and VEGF immunoreactivity, but no significant change in HO1 or astrogliosis. These novel findings reveal that CD163 deficiency has distinct temporal influences following ICH, with early beneficial properties but delayed injurious effects. While it is unclear why CD163 deficiency is initially beneficial, the late injurious effects are consistent with the key anti-inflammatory role of CD163 in the recovery phase of tissue damage.

Novel mutation in Sjogren-Larsson syndrome is associated with divergent neurologic phenotypes.

Sjögren-Larsson syndrome is an inherited disorder of lipid metabolism caused by mutations in the ALDH3A2 gene that codes for fatty aldehyde dehydrogenase, which results in accumulation of fatty aldehydes and alcohols and is characterized by ichthyosis, intellectual disability, and spastic diplegia/quadriplegia. The authors describe 2 unrelated Honduran patients who carried the same novel homozygous nonsense mutation (c.1309A>T, p.K437X) and ALDH3A2 DNA haplotype, but widely differed in disease severity. One patient exhibited spastic quadriplegia with unusual neuroregression, whereas the other patient had the usual static form of spastic diplegia with neurodevelopmental disabilities. Biochemical analyses showed a similar profound deficiency of fatty aldehyde dehydrogenase activity and impaired fatty alcohol metabolism in both patients' cultured fibroblasts. These results indicate that variation in the neurologic phenotype of Sjögren-Larsson syndrome is not strictly determined by the ALDH3A2 mutation or the biochemical defect as expressed in cultured fibroblasts, but by unidentified epigenetic/environmental factors, gene modifiers, or other mechanisms.

Expanding the neurologic phenotype of oculodentodigital dysplasia in a 4-generation Hispanic family.

We report a 4-generation Hispanic family with oculodentodigital dysplasia whose members were found to have typical phenotypic characteristics of this disorder, as well as a variable expression of neurologic manifestations in multiple generations ranging from a mild spastic gait to moderate to severe spastic tetraparesis/quadriplegia with epilepsy and an abnormal brain and spinal cord magnetic resonance imaging result. Gene testing documented a previously reported missense mutation in GJA1 (connexin 43) exon 2 (c.389T>C;p.I130T). Our evaluation not only expands the phenotypes associated with GJA1 gene mutations but also demonstrates that a great degree of variability in neurological defects can exist within a single family without evidence of genetic anticipation. A genotype-phenotype correlation between the p.I130T mutation and neurologic dysfunction appears more likely with the addition of this report's neurologic and GJA1 gene mutation findings. These findings expand the neurologic phenotype and prognosis and underscore the importance of counseling families with oculodentodigital dysplasia about the possibility of neurologic involvement.

Factores de riesgo asociados a la depresión en pacientes de la consulta dermatológica en dos hospitales de la ciudad de Barranquilla (Colombia) / Associated risk factors to depression in dermatology care at hospitals in the city of Barranquilla (Colombia)

objetivos: Analizar los factores personales y familiares asociados a la depresión en los pacientes dela consulta externa dermatológica del Hospital Universidad del Norte y del Hospital Nazareth de laciudad de Barranquilla (Colombia).Resultados: Se encontró una fuerte asociación entre la presencia de depresión y los antecedentespersonales y familiares de la enfermedad, la evolución prolongada de la enfermedad dermatológica, laspérdidas laborales y afectivas, la presencia de enfermedad concomitante y la disfuncionalidad familiar.Conclusión: Los resultados sugieren que la depresión en los pacientes con trastornos dermatológicosestá asociada a la influencia de factores similares a los encontrados en la población general, a lo cualse añade el tiempo de evolución de la enfermedad dermatológica y la disfuncionalidad familiar.

Objectives: To analyze the association between personal and familiar factors and the presence ofdepression in dermatologic patients that attend the outpatient service at the Hospital Universidaddel Norte and the Hospital Nazareth in Barranquilla.Methods: A cross sectional study was designed within a population of 339 dermatology outpatients.The sample was selected by systematic sampling methods. Cases were defined as having some degreeof depression diagnosed by the Hamilton Rating Scale for Depression and controls as those who werenegative. Individual factors studied were: sex, marriage and economic status, length of evolution ofdermatological disorder, former history of depression, loss of job and loss of loved ones. Familiar factorsanalyzed were family history of psychiatric disorders, depression and familiar function. Data analysiswas computerized and the prevalence odds ratios (POR) and their confidence intervals werecalculated. The significance tests for association utilized were Chi-squared and Fisher.Results: Depression was associated to: personal and familiar background of depression, length ofevolution of the dermatological problem ...