COVID-19 mortality in an area of northeast Brazil: epidemiological characteristics and prospective spatiotemporal modelling.

This study aimed to analyse the spatial-temporal distribution of COVID-19 mortality in Sergipe, Northeast, Brazil. It was an ecological study utilising spatiotemporal analysis techniques that included all deaths confirmed by COVID-19 in Sergipe, from 2 April to 14 June 2020. Mortality rates were calculated per 100 000 inhabitants and the temporal trends were analysed using a segmented log-linear model. For spatial analysis, the Kernel estimator was used and the crude mortality rates were smoothed by the empirical Bayesian method. The space-time prospective scan statistics applied the Poisson's probability distribution model. There were 391 COVID-19 registered deaths, with the majority among ⩾60 years old (62%) and males (53%). The most prevalent comorbidities were hypertension (40%), diabetes (31%) and cardiovascular disease (15%). An increasing mortality trend across the state was observed, with a higher increase in the countryside. An active spatiotemporal cluster of mortality comprising the metropolitan area and neighbouring cities was identified. The trend of COVID-19 mortality in Sergipe was increasing and the spatial distribution of deaths was heterogeneous with progression towards the countryside. Therefore, the use of spatial analysis techniques may contribute to surveillance and control of COVID-19 pandemic.

Risk clusters of COVID-19 transmission in northeastern Brazil: prospective space-time modelling.

This study aimed to analyse the trend and spatial-temporal clusters of risk of transmission of COVID-19 in northeastern Brazil. We conducted an ecological study using spatial and temporal trend analysis. All confirmed cases of COVID-19 in the Northeast region of Brazil were included, from 7 March to 22 May 2020. We used the segmented log-linear regression model to assess time trends, and the local empirical Bayesian estimator, the global and local Moran indexes for spatial analysis. The prospective space-time scan statistic was performed using the Poisson probability distribution model. There were 113 951 confirmed cases of COVID-19. The average incidence rate was 199.73 cases/100 000 inhabitants. We observed an increasing trend in the incidence rate in all states. Spatial autocorrelation was reported in metropolitan areas, and 178 municipalities were considered a priority, especially in the states of Ceará and Maranhão. We identified 11 spatiotemporal clusters of COVID-19 cases; the primary cluster included 70 municipalities from Ceará state. COVID-19 epidemic is increasing rapidly throughout the Northeast region of Brazil, with dispersion towards countryside. It was identified high risk clusters for COVID-19, especially in the coastal side.

Slow pyrolysis of Spirulina platensis for the production of nitrogenous compounds and potential routes for their separation.

The potential of microalgae Spirulina platensis to the production of nitrogenous compounds in liquid fraction via slow pyrolysis was evaluated. Aiming to identify the best condition which maximized liquid yield, the effects of operational conditions mass load, temperature, and heating rate were evaluated using Experimental Design and Response Surface Methodology techniques and optimized with Differential Evolution methodology. The composition of liquid fraction was analyzed by GC-MS and the effect of the same operational conditions in nitrogenous compounds formation was analyzed. The separation of nitrogenous compounds was evaluated by extraction and adsorption techniques. The results indicated that the heating rate significantly impacted both the liquid yield and the formation of the nitrogenous compounds. At optimal conditions, a maximum liquid yield of 64.59% was obtained. The extraction and adsorption processes showed to be promising routes for the purification of nitrogenous compounds, however, extraction was more selective to separate them.

Dairy production practices and associated risks for bovine vaccinia exposure in cattle, Brazil.

A cross-sectional serosurvey was performed to identify environmental features or practices of dairy farms associated with risk for exposure to vaccinia-like viruses in dairy cattle in Brazil. Sera from 103 cows from 18 farms in Minas Gerais state were examined for Orthopoxvirus-neutralizing antibodies. A database of 243 binary or multiple-selection categorical variables regarding the physical features and surrounding ecology of each property was obtained. Thirteen of 46 presumptive predictor variables were found to be significantly associated with Orthopoxvirus serostatus by univariate logistic regression methods. Use of teat sanitizer and having felids on the property were independently associated with virus exposure by multivariable analysis. Rodents have long been suspected of serving as maintenance reservoirs for vaccinia-like viruses in Brazil. Therefore, domestic felids are not only effective predators of small rodent pests, but also their urine can serve as a deterrent to rodent habitation in buildings such as stables and barns. These results corroborate previous evidence of the high significance of rodents in the Vaccinia virus transmission cycle, and they also raise questions regarding the common use of teat sanitizers in dairy production areas.

Whole-volume vs. segmental CT texture analysis of the liver to assess metachronous colorectal liver metastases.

PURPOSE: It is unclear whether changes in liver texture in patients with colorectal cancer are caused by diffuse (e.g., perfusional) changes throughout the liver or rather based on focal changes (e.g., presence of occult metastases). The aim of this study is to compare a whole-liver approach to a segmental (Couinaud) approach for measuring the CT texture at the time of primary staging in patients who later develop metachronous metastases and evaluate whether assessing CT texture on a segmental level is of added benefit. METHODS: 46 Patients were included: 27 patients without metastases (follow-up >2 years) and 19 patients who developed metachronous metastases within 24 months after diagnosis. Volumes of interest covering the whole liver were drawn on primary staging portal-phase CT. In addition, each liver segment was delineated separately. Mean gray-level intensity, entropy (E), and uniformity (U) were derived with different filters (σ0.5-2.5). Patients/segments without metastases and patients/segments that later developed metachronous metastases were compared using independent samples t tests. RESULTS: Absolute differences in entropy and uniformity between the group without metastases and the group with metachronous metastases group were consistently smaller for the segmental approach compared to the whole-liver approach. No statistically significant differences were found in the texture measurements between both groups. CONCLUSIONS: In this small patient cohort, we could not demonstrate a clear predictive value to identify patients at risk of developing metachronous metastases within 2 years. Segmental CT texture analysis of the liver probably has no additional benefit over whole-liver texture analysis.

Genetic divergence of sugarcane varieties collected in the region of Lavras, Minas Gerais, Brazil.

Genetic diversity among local accessions and varieties subsidize plant breeding programs, allowing the utilization of existing variability in plants that have already adapted to local climate conditions. An alternative to studying genetic variability is the study of diversity. The aim of this research was to study genetic diversity among sugarcane accessions and varieties used for the production of craft-distilled cachaça (distilled sugarcane alcohol) in the region of Lavras, Minas Gerais, Brazil. Using a one-way design, an experiment was conducted in the municipality of Perdões, Minas Gerais to evaluate 35 regional accessions derived from germplasm collection expeditions and four varieties. Using morphological descriptions of 46 multicategorical sugarcane characteristics, dissimilarity and Tocher cluster method analyses were performed. Based on the results, it was concluded that genetic diversity exists among the accessions evaluated for the target traits.

Is there a relationship between polyploidy and stressful environments? A case study of inselbergs in northeastern Brazil.

The aim of this study was to examine the hypothesis that plants with higher ploidy levels are selected by environments under more extremely stressful conditions than the same (or closely related) species from less rigorous terrestrial or epiphytic habitats. Therefore, we analyzed the chromosome numbers of 26 species belonging to 21 genera and 13 families of angiosperms that were collected on 4 inselbergs in Paraiba State, northeastern Brazil. In addition, 13 rupicolous species that grow in the study area, whose chromosome numbers were previously determined, were included in our analysis. Plant chromosome numbers varied between 2n = 12 in Callisia filiformis (Commelinaceae) to 2n = ca. 240 in Epidendrum cinnabarinum (Orchidaceae). The data as a whole do not support the hypothesis that inselberg species have greater levels of ploidy than terrestrial or epiphytic populations of the same or closely related species because these rupicolous species demonstrate chromosomal evolution patterns that are independent of the stressful environments of inselberg formations.

Optical biomarkers of serous and mucinous human ovarian tumor assessed with nonlinear optics microscopies.

BACKGROUND: Nonlinear optical (NLO) microscopy techniques have potential to improve the early detection of epithelial ovarian cancer. In this study we showed that multimodal NLO microscopies, including two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), third-harmonic generation (THG) and fluorescence lifetime imaging microscopy (FLIM) can detect morphological and metabolic changes associated with ovarian cancer progression. METHODOLOGY/PRINCIPAL FINDINGS: We obtained strong TPEF + SHG + THG signals from fixed samples stained with Hematoxylin & Eosin (H&E) and robust FLIM signal from fixed unstained samples. Particularly, we imaged 34 ovarian biopsies from different patients (median age, 49 years) including 5 normal ovarian tissue, 18 serous tumors and 11 mucinous tumors with the multimodal NLO platform developed in our laboratory. We have been able to distinguish adenomas, borderline, and adenocarcinomas specimens. Using a complete set of scoring methods we found significant differences in the content, distribution and organization of collagen fibrils in the stroma as well as in the morphology and fluorescence lifetime from epithelial ovarian cells. CONCLUSIONS/SIGNIFICANCE: NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for serous and mucinous ovarian tumors. The results provide a basis to interpret future NLO images of ovarian tissue and lay the foundation for future in vivo optical evaluation of premature ovarian lesions.

Ex vivo expansion of CD56+ NK and NKT-like lymphocytes from peripheral blood mononuclear cells of patients with ovarian neoplasia.

Methods for ex vivo expansion of natural killer (NK) cells have allowed obtaining enough numbers of human NK cells for clinical trials. However, the evaluation of these methods has been mostly limited to haematological malignancies. This study aimed at evaluating a method for selective expansion of NK cells when applied in peripheral blood mononuclear cells (PBMC) of patients with ovarian neoplasia. PBMC from 13 volunteer patients with ovarian neoplasia, seven benign and six malignant tumours, were cultured in CellGro medium supplemented with anti-CD3 (9-10 initial days), IL-2 and foetal bovine serum for 21 days. The resulting effector cells were evaluated for their phenotype, cytotoxicity and cytokine secretion. PBMC cultures resulted in multiple populations (NK, NKT and T) of effector cells, enriched with CD56(+) lymphocytes. NK cells from patients with benign and malignant ovarian neoplasia were expanded 139.6 ± 63.4 and 82.7 ± 25.3-fold, respectively, being the largest lymphocyte subtype among CD56(+) population. Effector cells expanded from patients with malignant ovarian neoplasia had higher proportion of T lymphocytes and altered cytokine production patterns, characterized by lower INF-γ, TNF-α and higher IL-4, compared with patients with benign ovarian neoplasia. Effector cells were cytotoxic against K562 and OVCAR3 cell lines. Cytotoxicity was significantly higher (P < 0.05) using magnetically separated CD56(+) effector cell fractions compared with CD56-deprived ones. The present study demonstrates the feasibility of the culture system employed to generate effector cells, enriched with CD56(+) lymphocytes, from PBMC of patients with ovarian neoplasia. NK cells were the largest lymphocyte subtype among the CD56(+) population and the main variable among the final effector cell preparation affecting target cell killing.

Stromal cells play a role in cervical cancer progression mediated by MMP-2 protein.

Metalloproteinases, especially metalloproteinase-2 (MMP-2), are known for their role in the degradation of the extracellular matrix. Nevertheless, a thorough understanding of MMP-2 expression in neoplastic lesions of the uterine cervix has yet to be accomplished. This study aimed to analyze the MMP-2 expression in cervical intraepithelial neoplasia III (CIN3) and in cervical squamous cell carcinoma, in tumor cells and adjacent stromal cells. MMP-2 expression was assessed by an immunohistochemical technique. MMP-2 expression was greater in the stromal cells of invasive carcinomas than in CIN3 (p < 0.0001). MMP-2 expression in stromal cells correlates with the clinical stage, gradually increasing as the tumor progresses (p = 0.04). This study corroborates that stromal cells play an important role in tumor invasion and progression, mediated by the progressive enhancement of MMP-2 expression from CIN3 to advanced invasive tumor. The intense MMP-2 expression most probably is associated with poor tumor prognosis.

Endocervical glandular cell abnormalities in conventional cervical smears: evaluation of the performance of cytomorphological criteria and HPV testing in predicting neoplasia.

OBJECTIVE: To analyse the correlation between cytomorphological criteria in smears with atypical glandular cells (AGC) or adenocarcinoma in situ (AIS) and human papillomavirus (HPV) reflex test results with different neoplastic histological diagnoses, particularly to distinguish between glandular and squamous neoplasia. METHODS: A series of 155 women with glandular abnormalities in their conventional cervical smears was included: 106 with AGC, 35 with AGC associated with high-grade squamous intraepithelial lesion (HSIL) and 14 with AIS. Two reviewers evaluated 35 cytomorphological criteria and hybrid capture II (HCII) was performed in all cases. Colposcopy was carried out in all cases and biopsy in 126/155. For statistical purposes, predictive values and odds ratio (OR) were calculated, followed by chi-square automatic interaction detection. RESULTS: Histology detected 56 cases of squamous and 17 of glandular intraepithelial or invasive neoplasia. Predictive values of the papillary groups and feathering criteria for glandular neoplasia were, respectively, 80.0% and 73.3%. Feathering was the criterion with the highest OR for distinguishing glandular from squamous neoplasia and also for distinguishing between glandular and non-neoplastic diagnosis. Rosettes and pseudostratified strips did not perform as well. Multivariant Classification and Regression Trees analysis identified feathering as the best criterion for distinguishing between glandular, squamous and non-neoplastic diagnoses regardless of HPV status. CONCLUSIONS: Feathering was the best criterion for predicting glandular neoplasia.

Expression of p16 and cervical infection with high-risk human papillomavirus are not related to p53 activity in cervical intraepithelial neoplasia.

The objective of the study was to investigate the expression of p53 and p16(INK4a) in cervical intraepithelial neoplasia (CIN) and their relation with disease severity and high-risk human papillomavirus (HR-HPV) status. A series of 125 women with previous positive Pap smear were selected for this cross-sectional study. All patients underwent gynecological examination, including colposcopy. Specimens for Pap smears, Hybrid Capture 2 (HC2) test, and pathologic analysis were obtained. After evaluation of CIN grade, immunohistochemical detection of proteins p53 and p16(INK4a) was performed on paraffin-embedded sections. The extent of immunoexpression of both proteins was analyzed in relation to CIN grade and HR-HPV status. CIN was graded as 1 in 21, 2 in 17, and 3 in 87 specimens. p16(INK4a) positivity (at least 5% of epithelial cells stained) was found in 99 of 125 cases (79.2%) and was significantly higher in high-grade lesions as compared to low-grade CIN (P < 0.001). The expression of p53 did not differ across histologic strata. Protein expression neither of p16(INK4a) nor of p53 correlated with HR-HPV status. Expression of p16(INK4a) was not related with that of p53. Our study gives further support to previous findings of strong association of p16(INK4a) immunostaining with severity of epithelial atypia, but this protein may not be considered a predictor of HR-HPV status determined with HC2. By contrast, immunoexpression of p53 was related neither to CIN grade nor to HR-HPV status.

Cell proliferation activity unrelated to COX-2 expression in ovarian tumors.

The objective of this study was to assess the expression of Cyclooxygenase-2 (COX-2) and cell proliferation activity (Ki67 expression) in benign, borderline, and malignant serous and mucinous ovarian tumors. Expression of COX-2 and Ki67 proteins were evaluated by immunohistochemistry, in paraffin-embedded sections of ovarian epithelial tumors. The study included 113 serous (67 benign, 15 borderline, and 31 malignant) and 85 mucinous (48 benign, 28 borderline, and 9 malignant) tumors, removed from women who underwent laparotomy between January 1997 and December 2003. From benign to malignant tumors, there was a progressive positive trend in COX-2 expression in both serous and mucinous tumors, more evident in mucinous ones (P < 0.001). Comparing histologic types, COX-2 expression was more prominent in serous than in mucinous benign tumors (P < 0.01), but this difference was not significant in the borderline (P= 0.11) or malignant categories (P= 0.71). There was a progressive Ki67 positivity in line with the tumor histologic gradient for both serous (P < 0.01) and mucinous lesions (P < 0.01), but this increasing expression did not correlate with COX-2 expression in the present series (P= 0.78). There was a higher COX-2 expression in serous ovarian adenomas than in mucinous ones. COX-2 positivity increases in line with the morphologic gradient, from benign to malignant in both histologic types, but it was more prominent in mucinous lesions, pointing to different oncogenic pathways related to different histologic types. A correlation between the expression of COX-2 and Ki67 was not found, suggesting that COX-2 may be required for carcinogenesis, but this pathway is not responsible for cell proliferation in ovarian tumors.

Molecular imaging in hereditary forms of parkinsonism.

The development of in vivo molecular imaging to evaluate the dopamine (DA) system with positron-emission tomography and single photon emission computed tomography has been of key importance on monitoring in vivo nigrostriatal neuronal loss in Parkinson's disease (PD), mostly through assessments of pre- and post-synaptic DA receptors. The discoveries of genes related to hereditary forms of parkinsonism (PARK1, PARK2, PARK6, PARK7 and PARK8) have increased our understanding either of distinct subtypes of clinical expression in PD or its etiology. This article revises current data on molecular neuroimaging of genetic forms of parkinsonism comparing and contrasting its main features with the classical sporadic forms. Awareness of the spectrum variance in the genotype and its respective PD phenotype are useful to distinguish different pathophysiological mechanisms of PD.

Prediction of high-grade cervical disease with human papillomavirus detection in women with glandular and squamous cytologic abnormalities.

The objective of this study was to assess whether human papillomavirus (HPV) detection with hybrid capture II (HC II) can help predict the presence and the nature, glandular or squamous, of histologic cervical lesions in women referred due to atypical glandular cells (AGC) or high-grade squamous intraepithelial lesion (HSIL). A total of 247 women were included. Referral Pap smears comprised AGC (51 cases), AGC plus HSIL (28 cases), adenocarcinoma in situ (10 cases), and HSIL (158 cases). All patients were tested for high-risk HPV with HC II and had a histologic assessment of their cervix. Histologic analysis showed 38 women with (15.3%) cervicitis, 194 with (75.5%) squamous lesions, and 15 with (9.2%) glandular neoplasia. The overall rate of high-risk HPV detection was 77%. Almost 70% of AGC-HPV-negative patients did not have a pathologically proven cervical neoplasia, whereas 76% of women with AGC-HPV-positive result were diagnosed with a squamous or glandular neoplasia. Most (95%) of the lesions in patients with AGC-HSIL were of squamous nature, and HPV detection did not contribute to their differentiation from glandular lesions. We conclude that in women with AGC, HPV positivity strongly correlated with the presence of glandular or squamous cervical lesion but did not help distinguishing women with squamous from those with glandular neoplasia.

PTEN and p53 expression in primary ovarian carcinomas: immunohistochemical study and discussion of pathogenetic mechanisms.

Proapoptotic molecules have been studied in epithelial ovarian neoplasms as possible indicators of the pathogenetic pathways, as targets for new therapeutic approaches, and as prognostic markers. PTEN and p53 are proteins that have many different regulatory functions, including apoptosis. We have studied their immunohistochemical expression in 70 cases of primary ovarian carcinomas (26 serous, 27 endometrioid, and 17 mucinous) and compared the results with morphologic parameters (histologic grade, subtype) and clinical data (age, stage, tumor size). Statistical analyses showed a significantly higher expression of p53 in histologically high-grade tumors (grades 2 and 3), mainly of the serous subtype. A statistical tendency of higher expression of p53 in older patients (P= 0.08) was also observed. The loss of expression of PTEN was significantly more frequent in grade 1 endometrioid adenocarcinomas. These markers did not show association with volume or stage of the tumor. p53 is associated with serous carcinoma, loss of differentiation, and older patients, whereas PTEN inactivation is an early event in carcinogenesis of the endometrioid subtype, as observed in type I endometrial carcinoma. Our results are in keeping with different pathogenetic pathways in subtypes of ovarian carcinoma, prompting the search for new strategies of prevention and treatment.

Atypical glandular cells: criteria to discriminate benign from neoplastic lesions and squamous from glandular neoplasia.

OBJECTIVE: To evaluate the presence of some criteria in cervical smears with atypical glandular cells and their correlation with histological patterns to identify pre-neoplastic and neoplastic lesions. METHODS: Seventy-three women referred with an atypical glandular cell smear, who had undergone conization or hysterectomy, were included in this study. Referral Pap smears were reviewed using the set of 27 cyto-morphological criteria that was correlated with the histological diagnosis. RESULTS: Histological results showed intraepithelial or invasive neoplasia in 35 (48%) cases and benign lesions in 38 (52%) cases. After logistic regression and decision tree analysis an increased nuclear/cytoplasmic ratio and the presence of dyskeratotic cells were strongly associated with intraepithelial or invasive neoplasia and the differential cyto-morphological criteria for glandular lesions were decreased cytoplasm, irregular nuclear membranes and the presence of nucleoli. CONCLUSION: The analysis of individual cyto-morphological criteria can better predict intraepithelial or invasive neoplasia and differentiate glandular from squamous lesions.

Nuclear alterations of cells and atypical metaplastic cells in cervical smears are predictive criteria of high-grade cervical intraepithelial neoplasia.

Diagnoses based on the screening of cervical smears show low interobserver reproducibility and are frequently discordant with the final histological diagnosis. The aim of this study was to identify which of the cytomorphologic criteria used in the screening of cervical smears were most predictive of the histopathological grades of cervical intraepithelial neoplasia. The abnormal cervical smears of 206 women were reviewed blindly according to 22 pre-established cytomorphological criteria. Colposcopic evaluation was carried out in all cases. The marked presence of several nuclear criteria frequently found together in the same smear was associated with high grade intraepithelial neoplasia regardless of the presence of any other criteria. On the other hand, when the nuclear criteria were less evident, the cluster of criteria related to metaplastic cells was predictive of a diagnosis of high-grade intraepithelial neoplasia. Focusing on selected cytological criteria can aggregate predictive value to cervical smear diagnoses.