Amoebicidal effect of synthetic indoles against Acanthamoeba spp.: a study of cell death.

Organic and synthetic chemistry plays a crucial role in drug discovery fields. Moreover, chemical modifications of available molecules to enhance their efficacy, selectivity and safety have been considered as an attractive approach for the development of new bioactive agents. Indoles, a versatile group of natural heterocyclic compounds, have been widely used in pharmaceutical industry due to their broad spectrum of activities including antimicrobial, antitumoral and anti-inflammatory among others. Herein, we report the amoebicidal activity of different indole analogs on Acanthamoeba castellanii Neff. Among the 40 tested derivatives, eight molecules were able to inhibit this protistan parasite. The structure-activity relationship (SAR) analysis of their anti-Acanthamoeba activity would suggest that a carboxylation of C-3 position and the incorporation of halogen as chlorine/fluorine would enhance their biological profile, presumably by increasing their lipophilicity and therefore their ability to cross the cell membrane. Fluorescence image base system was used to investigate the effect of indole 6o c-6 on the cytoskeleton network and various programmed cell death features. We were able to highlight that the methyl 6-chloro-1H-indole-3-carboxylate could induce program cell death by the mitochondrial dysfunction.

A General and Scalable Synthesis of Polysubstituted Indoles.

A consecutive 2-step synthesis of N-unprotected polysubstituted indoles bearing an electron-withdrawing group at the C-3 position from readily available nitroarenes is reported. The protocol is based on the [3,3]-sigmatropic rearrangement of N-oxyenamines generated by the DABCO-catalyzed reaction of N-arylhydroxylamines and conjugated terminal alkynes, and delivers indoles endowed with a wide array of substitution patterns and topologies.

Recent Advances in the Synthesis of 2H-Pyrans.

In this review, we discuss the nature of the different physicochemical factors affecting the valence isomerism between 2H-pyrans (2HPs) and 1-oxatrienes, and we describe the most versatile synthetic methods reported in recent literature to access to 2HPs, with the only exception of 2HPs fused to aromatic rings (i.e., 2H-chromenes), which are not included in this review.

Agreement Between Preoperative Endometrial Sampling and Surgical Specimen Findings in Endometrial Carcinoma.

OBJECTIVES: The aims of the study were to evaluate agreement between preoperative endometrial samples and surgical specimens in endometrial carcinoma and to correlate this agreement with sample and patient characteristics. METHODS: Patients who received primary surgical treatment for endometrial carcinoma at a tertiary care center and had undergone preoperative endometrial sampling were included. Medical records were reviewed to collect information from pathology reports and data on patient characteristics. RESULTS: The study sample comprised 166 patients (mean age, 64.6 years). The histological results of the biopsies were the following: endometrioid cancer (n = 118), nonendometrioid tumor (n = 38), and hyperplasia (n = 10). The agreement rates were 93.2% for endometrioid and 68.9% for nonendometrioid tumors, with a κ coefficient of 0.73 for tumor cell type. Tumor International Federation of Gynecology and Obstetrics (FIGO) grade was distributed as follows: 37.1% G1, 35.7% G2, and 27.1% G3, with agreement rates of 61.5%, 56%, and 78.9%, respectively. The overall κ coefficient for FIGO grading was 0.46. Only 1.9% of the tumors originally classified as G1 were upgraded to G3, whereas 16% of G2 lesions were upgraded. There was no significant difference in agreement rates for tumor cell type and FIGO grade in relation to any of the studied variables, except that biopsy specimens weighing more than 3 g had significantly better agreement in FIGO grading (P = 0.040). CONCLUSIONS: Preoperative biopsy has suboptimal accuracy for prediction of characteristics in the definitive surgical specimen. Caution must be taken when using preoperative information to determine extent of surgical resection, due to the risk of understaging. Additional information must be combined with the biopsy data to help in the decision-making process.

Carcinogenesis of Vulvar Lesions: Morphology and Immunohistochemistry Evaluation.

OBJECTIVES: The aim of the study was to assess the 2 pathways of vulvar carcinogenesis and correlate immunohistochemical expression of p53 with histopathological findings. MATERIALS AND METHODS: This cross-sectional study included 76 cases. Patients were classified according to the 2004 International Society for the Study of Vulvovaginal Disease Terminology, followed by a review of clinical records and immunohistochemical staining for p53. RESULTS: Fifteen cases were in the human papillomavirus (HPV)-associated pathway (12 cases of usual vulvar intraepithelial neoplasia [VIN] and 3 of warty squamous cell carcinoma [SCC]), and 13 cases were in the HPV-independent pathway (5 cases of differentiated VIN and 8 of keratinizing SCC). Significant differences in p53 expression were observed between the 2 pathways of carcinogenesis: in the lesions related to the HPV-independent pathway, the percentage of p53-positive cells was greater (>25%, p < .001), and the staining pattern was basal (extending into the middle layer) in differentiated VIN and diffuse or infiltrative in warty SCC (p < 0.001). In the lesions HPV-associated pathway, p53 staining was less extensive (≤10% of cells, p < 0.001) and followed basal pattern in usual VIN, whereas warty SCCs were negative for p53 (p < 0.001). CONCLUSIONS: Unique patterns of histological appearance and p53 expression can separate vulvar lesions into 2 distinct pathways of carcinogenesis. We propose that p53 immunohistochemistry may be performed simultaneously with histopathological examination in all cases of VIN and vulvar SCC, because it would aid in definition of the pathway of carcinogenesis and thus enable better clinical follow-up of patients with these conditions.

Telomere length assessment and molecular characterization of TERT gene promoter in periampullary carcinomas.

Genetic and epigenetic alterations of the telomere maintenance machinery like telomere length and telomerase reverse transcriptase (encoded by TERT gene) are reported in several human malignancies. However, there is limited knowledge on the status of the telomere machinery in periampullary carcinomas (PAC) which are rare and heterogeneous groups of cancers arising from different anatomic sites around the ampulla of Vater. In the current study, we investigated the relative telomere length (RTL) and the most frequent genetic and epigenetic alterations in the TERT promoter in PAC and compared it with tumor-adjacent nonpathological duodenum (NDu). We found shorter RTLs (1.27 vs 1.33, P = 0.01) and lower TERT protein expression (p = 0.04) in PAC tissues as compared to the NDu. Although we did not find any mutation at two reactivating hotspot mutation sites of the TERT promoter, we detected polymorphism in 45% (9/20) of the cases at rs2853669 (T > C). Also, we found a hypermethylated region in the TERT promoter of PACs consisting of four CpGs (cg10896616 with Δß 7%; cg02545192 with Δß 9%; cg03323598 with Δß 19%; and cg07285213 with Δß 15%). In conclusion, we identified shorter telomeres with DNA hypermethylation in the TERT promoter region and lower TERT protein expression in PAC tissues. These results could be used further to investigate molecular pathology and develop theranostics for PAC.

Autopsy in a Neonatal Intensive Care Unit: pathological and clinical agreement.

OBJECTIVES: To evaluate neonatal autopsy rates at a tertiary hospital in southern Brazil ascertain the level of agreement between premortem and postmortem diagnosis. METHODS: The authors reviewed all neonatal autopsies performed over a 10-year period and described the percentage of neonates who died and underwent autopsy. The authors tested for agreement between autopsy findings and the cause of death as defined by the neonatologist. Agreement between clinical diagnosis and autopsy findings was classified using the modified Goldman criteria. Additional findings at autopsy were grouped by organ system. Linear regression and multiple comparisons were used for statistical analyses. RESULTS: During the study period, 382 neonates died at the Neonatal Intensive Care Unit (NICU). Consent to perform an autopsy was obtained for 73 (19.1%). The complete agreement between autopsy findings and the neonatologist's premortem diagnosis was found in 48 patients (65.8%). Additional findings were obtained at autopsy in 25 cases (34.2%). In 5 cases (6.9%), the autopsy findings contributed to subsequent genetic counseling. Seven autopsies (9.6%) revealed a diagnosis that would have changed patient management if established premortem. The autopsy rate increased by an average of 1.87% each year. CONCLUSION: Despite a high level of agreement between clinical diagnosis and pathological findings, autopsies provided relevant data regarding the cause of death, providing additional clinical information to neonatologists and allowing genetic counseling of family members.

Dynamic Hydroxyl-Yne Reaction with Phenols.

Dynamic Covalent Chemistry (DCvC) has gained increasing importance in supramolecular chemistry and materials science. Herein we prove the dynamic nature of the exchange between phenols and vinyl ethers. Exchange is fast at room temperature and under mild conditions. The equilibrium constants and the electronic effect of the phenol substituents were calculated. This novel incorporation to the DCvC toolbox could be quite useful, and as a proof it was used for the synthesis of a responsive molecular cage.

Sarcina ventriculi a rare pathogen.

Sarcina ventriculi is a gram-positive bacterium, able to survive in extreme low pH environment. It's first description dates from 1842, by John Goodsir. Since then, just a few cases have been reported. In veterinary medicine, especially in ruminants, it causes bloating, vomiting, gastric perforation and death of the animal. It is commonly associated with delayed gastric emptying or obstruction to gastric outlet, although it's pathogenicity in humans is not fully understood. We report two cases with identification of the bacteria in gastric specimens stained with hematoxylin-eosin staining, in different clinical settings. The first patient is a young female patient, presenting cardiac arrest and death after gastric perforation and the second patient an adult male presenting with gastric adenocarcinoma, treated with partial gastrectomy followed by adjuvant chemoradiation. In our literature review, we identified forty-five cases reporting Sarcina ventriculi appearance, with a sudden increase since 2010.

Short and Modular Synthesis of Substituted 2-Aminopyrroles.

We herein describe a simple and metal-free domino methodology to synthesize 2-aminopyrroles from alkynyl vinyl hydrazides. The domino reaction involves a novel propargylic 3,4-diaza-Cope rearrangement and a tandem isomerization/5-exo-dig N-cyclization reaction. By using this approach, a number of 2-aminopyrroles with diverse substituents have been prepared.

Use of clomiphene citrate protocol for controlled ovarian stimulation impairs endometrial maturity.

INTRODUCTION: Despite recent advances in assisted reproduction techniques and recent knowledge regarding embryo and endometrium quality, implantation and birth rates remain low. The objective of this study was to investigate whether clomiphene citrate alters endometrial maturation in infertile patients. METHODS: In a prospective self-matched cohort study, we assessed the ovulation of women in spontaneous and stimulated cycles (with clomiphene citrate). We determined the ovulation day by ultrasound scanning. In both cycles, we took four blood samples (BS1 - at early proliferative phase, BS2 - at mid proliferative phase, BS3 - after ovulation and BS4 - at mid luteal phase) to determine the serum concentrations of FSH, LH, estradiol and progesterone. We retrieved an endometrial biopsy five days after ovulation, followed by blinded analysis and classification according to Noyes criteria, in both cycles. RESULTS: Twenty-two participants completed the study. There were significant differences in FSH BS3 (p=0.001), in LH BS3 and BS4 (p<0.001 and p=0.049, respectively), in estradiol BS2, BS3 and BS4 (p<0.001, p=0.024 and p<0.001, respectively) and in progesterone BS3 and BS4 (p=0.028 and p<0.001, respectively). Considering Noyes criteria, there was a one-day delay when comparing the stimulated cycle with the spontaneous cycle (p=0.004), and a two-day delay when comparing the stimulated cycle with the biopsy day. CONCLUSION: This study indicates that ovarian stimulation with clomiphene citrate delays the endometrial maturity, and could possibly impair the implantation process due to asynchrony.

Design, Synthesis and Characterization of HIV-1 CA-Targeting Small Molecules: Conformational Restriction of PF74.

Small molecules targeting the PF74 binding site of the HIV-1 capsid protein (CA) confer potent and mechanistically unique antiviral activities. Structural modifications of PF74 could further the understanding of ligand binding modes, diversify ligand chemical classes, and allow identification of new variants with balanced antiviral activity and metabolic stability. In the current work, we designed and synthesized three series of PF74-like analogs featuring conformational constraints at the aniline terminus or the phenylalanine carboxamide moiety, and characterized them using a biophysical thermal shift assay (TSA), cell-based antiviral and cytotoxicity assays, and in vitro metabolic stability assays in human and mouse liver microsomes. These studies showed that the two series with the phenylalanine carboxamide moiety replaced by a pyridine or imidazole ring can provide viable hits. Subsequent SAR identified an improved analog 15 which effectively inhibited HIV-1 (EC50 = 0.31 µM), strongly stabilized CA hexamer (ΔTm = 8.7 °C), and exhibited substantially enhanced metabolic stability (t1/2 = 27 min for 15 vs. 0.7 min for PF74). Metabolic profiles from the microsomal stability assay also indicate that blocking the C5 position of the indole ring could lead to increased resistance to oxidative metabolism.

Calcium Signaling Alterations Caused by Epigenetic Mechanisms in Pancreatic Cancer: From Early Markers to Prognostic Impact.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with high mortality rates. PDAC initiation and progression are promoted by genetic and epigenetic dysregulation. Here, we aimed to characterize the PDAC DNA methylome in search of novel altered pathways associated with tumor development. We examined the genome-wide DNA methylation profile of PDAC in an exploratory cohort including the comparative analyses of tumoral and non-tumoral pancreatic tissues (PT). Pathway enrichment analysis was used to choose differentially methylated (DM) CpGs with potential biological relevance. Additional samples were used in a validation cohort. DNA methylation impact on gene expression and its association with overall survival (OS) was investigated from PDAC TCGA (The Cancer Genome Atlas) data. Pathway analysis revealed DM genes in the calcium signaling pathway that is linked to the key pathways in pancreatic carcinogenesis. DNA methylation was frequently correlated with expression, and a subgroup of calcium signaling genes was associated with OS, reinforcing its probable phenotypic effect. Cluster analysis of PT samples revealed that some of the methylation alterations observed in the Calcium signaling pathway seemed to occur early in the carcinogenesis process, a finding that may open new insights about PDAC tumor biology.

Performance of cardiopulmonary resuscitation feedback systems in a long-distance train with distributed traction.

BACKGROUND: Out-of-hospital cardiac arrest is common in public locations, including public transportation sites. Feedback devices are increasingly being used to improve chest-compression quality. However, their performance during public transportation has not been studied yet. OBJECTIVE: To test two CPR feedback devices representative of the current technologies (accelerometer and electromag- netic-field) in a long-distance train. METHODS: Volunteers applied compressions on a manikin during the train route using both feedback devices. Depth and rate measurements computed by the devices were compared to the gold-standard values. RESULTS: Sixty-four 4-min records were acquired. The accelerometer-based device provided visual help in all experiments. Median absolute errors in depth and rate were 2.4 mm and 1.3 compressions per minute (cpm) during conventional speed, and 2.5 mm and 1.2 cpm during high speed. The electromagnetic-field-based device never provided CPR feedback; alert messages were shown instead. However, measurements were stored in its internal memory. Absolute errors for depth and rate were 2.6 mm and 0.7 cpm during conventional speed, and 2.6 mm and 0.7 cpm during high speed. CONCLUSIONS: Both devices were accurate despite the accelerations and the electromagnetic interferences induced by the train. However, the electromagnetic-field-based device would require modifications to avoid excessive alerts impeding feedback.

Synthesis and Utility of 2,2-Dimethyl-2 H-pyrans: Dienes for Sequential Diels-Alder/Retro-Diels-Alder Reactions.

The practical use of 2,2-dimethyl-2 H-pyrans as electron-rich dienes in sequential Diels-Alder/retro-Diels-Alder (DA/rDA) domino processes to generate aromatic platforms has been demonstrated. Different polysubstituted alkyl 2-naphthoates have been synthesized by the DA/rDA reaction of benzynes and 2,2-dimethyl-2 H-pyrans. The use of other activated alkynes allows the access of substituted alkyl benzoate derivatives.

Autopsy in a Neonatal Intensive Care Unit: pathological and clinical agreement

Abstract Objectives To evaluate neonatal autopsy rates at a tertiary hospital in southern Brazil ascertain the level of agreement between premortem and postmortem diagnosis. Methods The authors reviewed all neonatal autopsies performed over a 10-year period and described the percentage of neonates who died and underwent autopsy. The authors tested for agreement between autopsy findings and the cause of death as defined by the neonatologist. Agreement between clinical diagnosis and autopsy findings was classified using the modified Goldman criteria. Additional findings at autopsy were grouped by organ system. Linear regression and multiple comparisons were used for statistical analyses. Results During the study period, 382 neonates died at the Neonatal Intensive Care Unit (NICU). Consent to perform an autopsy was obtained for 73 (19.1%). The complete agreement between autopsy findings and the neonatologist's premortem diagnosis was found in 48 patients (65.8%). Additional findings were obtained at autopsy in 25 cases (34.2%). In 5 cases (6.9%), the autopsy findings contributed to subsequent genetic counseling. Seven autopsies (9.6%) revealed a diagnosis that would have changed patient management if established premortem. The autopsy rate increased by an average of 1.87% each year. Conclusion Despite a high level of agreement between clinical diagnosis and pathological findings, autopsies provided relevant data regarding the cause of death, providing additional clinical information to neonatologists and allowing genetic counseling of family members.

Misdiagnosis of Streptococcus gallolyticus endocarditis.

Death certificate inaccuracy is of major concern both in the public health domain and in individual health care, since it may yield untruthful data on the incidence, prevalence, and lethality of medical entities, and may hamper prophylactic measures among those who share, with the deceased, the common genetic, environmental, or behavioral risk factors. An effective way to settle this haziness relies on the increase of autopsy performance, increasing manifold the exactitude as well as facing surprising diagnoses. In this report, the authors present the case of a middle-aged woman who sought medical care because of back pain accompanied by weight loss. She died suddenly and unexpectedly in the Emergency Room. In this case, due to the unusual clinical presentation and the patient's unexpected death, the causa mortis would not have been elucidated if the autopsy had not been undertaken.

Sarcina ventriculi a rare pathogen

Sarcina ventriculi is a gram-positive bacterium, able to survive in extreme low pH environment. It's first description dates from 1842, by John Goodsir. Since then, just a few cases have been reported. In veterinary medicine, especially in ruminants, it causes bloating, vomiting, gastric perforation and death of the animal. It is commonly associated with delayed gastric emptying or obstruction to gastric outlet, although it's pathogenicity in humans is not fully understood. We report two cases with identification of the bacteria in gastric specimens stained with hematoxylin-eosin staining, in different clinical settings. The first patient is a young female patient, presenting cardiac arrest and death after gastric perforation and the second patient an adult male presenting with gastric adenocarcinoma, treated with partial gastrectomy followed by adjuvant chemoradiation. In our literature review, we identified forty-five cases reporting Sarcina ventriculi appearance, with a sudden increase since 2010.

Cranial vault reconstruction with bone morphogenetic protein, calcium phosphate, acellular dermal matrix, and calcium alginate in mice.

PURPOSE: To evaluate experimental cranial vault reconstructions, by combining bone morphogenetic protein type 2 (BMP-2) and different matrices. METHODS: Fourty-nine animals were initially included (seven per group). We designed an experimental, open, prospective and comparative study, divided in seven groups: 1 - BMP-2+calcium phosphate (BT); 2 - BMP-2+acellular dermal matrix (BM); 3 - BMP-2+calcium alginate (BA); 4 - TCP; 5 - MDM; 6 - ALG; 7 - Bone autograft (BAG). A bone failure was created in left parietal bone of adult male mice. At the same procedure reconstruction was performed. After five weeks, animals were sacrificed, and reconstruction area was removed to histological analysis. After exclusion due to death or infection, thirty-eight animals were evaluated (BT=5; BM=6; BA=6; TCP=7; MDM=3; ALG=6; BAG=5). RESULTS: A higher incidence of infection has occurred in MDM group (57%, P=0.037). In cortical fusion, groups BAG, TCP, and BMP-2+TCP (BT) obtained the best scores, comparing to the others (P=0.00846). In new bone formation, groups BT, BAG, and TCP have presented the best scores (P=0.00835). When neovascularization was considered, best groups were BMP-2+MDM (BM), BMP-2+ALG (BA), TCP, and MDM (P=0.001695). BAG group was the best in bone marrow formation, followed by groups BT and TCP (P=0.008317). CONCLUSIONS: Bone morphogenetic protein type 2 increased bone regeneration in experimental skull reconstruction, especially when combined to calcium phosphate. Such association was even comparable to bone autograft, the gold-standard treatment, in some histological criteria.

Cranial vault reconstruction with bone morphogenetic protein, calcium phosphate, acellular dermal matrix, and calcium alginate in mice

PURPOSE: To evaluate experimental cranial vault reconstructions, by combining bone morphogenetic protein type 2 (BMP-2) and different matrices. METHODS: Fourty-nine animals were initially included (seven per group). We designed an experimental, open, prospective and comparative study, divided in seven groups: 1 - BMP-2+calcium phosphate (BT); 2 - BMP-2+acellular dermal matrix (BM); 3 - BMP-2+calcium alginate (BA); 4 - TCP; 5 - MDM; 6 - ALG; 7 - Bone autograft (BAG). A bone failure was created in left parietal bone of adult male mice. At the same procedure reconstruction was performed. After five weeks, animals were sacrificed, and reconstruction area was removed to histological analysis. After exclusion due to death or infection, thirty-eight animals were evaluated (BT=5; BM=6; BA=6; TCP=7; MDM=3; ALG=6; BAG=5). RESULTS: A higher incidence of infection has occurred in MDM group (57%, P=0.037). In cortical fusion, groups BAG, TCP, and BMP-2+TCP (BT) obtained the best scores, comparing to the others (P=0.00846). In new bone formation, groups BT, BAG, and TCP have presented the best scores (P=0.00835). When neovascularization was considered, best groups were BMP-2+MDM (BM), BMP-2+ALG (BA), TCP, and MDM (P=0.001695). BAG group was the best in bone marrow formation, followed by groups BT and TCP (P=0.008317). CONCLUSIONS: Bone morphogenetic protein type 2 increased bone regeneration in experimental skull reconstruction, especially when combined to calcium phosphate. Such association was even comparable to bone autograft, the gold-standard treatment, in some histological criteria. .