Dietary Lysine Requirements of Older Adults Stratified by Age and Sex.

BACKGROUND: Current recommendation for lysine in older adults, 30 mg/kg/d, is based on young adult data. Evidence suggests that amino acid requirements may differ between young and old adults with both sex and age having an effect in the elderly. OBJECTIVES: This study aimed to define the lysine requirements in healthy older adults using the indicator amino acid oxidation (IAAO) method with L-[1-13C] phenylalanine as the indicator and to compare the derived estimates based on age: 60-69 y and >70 y. METHODS: Fourteen healthy males and 16 healthy females [>60 y, body mass index (BMI) = 26.3 kg/m2] were randomly assigned to receive 3-7 lysine intakes from 10 to 80 mg/kg/d. Subjects were adapted to a standard liquid diet providing 1.0 g/kg/d protein and adequate energy, for 2 d, with indicator oxidation measurements performed on day 3. The rate of release of 13CO2 from the oxidation of L-[1-13C] phenylalanine was measured in breath. A 2-phase linear mixed-effect model, and parametric bootstrap were used to determine mean lysine requirements and the 95% confidence intervals (CIs). The overlap of the 95% CI between the 2 age groups were used to compare the requirement estimates. The null hypothesis was accepted if the interval contained zero. RESULTS: The mean and upper 95% CI of the lysine requirement for females were 32.9 and 40.9 and 46.2 and 53.7 mg/kg/d for those aged 60-69 y and >70 y, respectively. The mean and upper 95% CI of the lysine requirement for the 2 groups of males were not different so was combined to yield a mean and 95% CI of 32.2 and 38.2 mg/kg/d. CONCLUSIONS: To our knowledge, this is the first study to report on the lysine requirement in adults aged >60 y. These results provide a basis from which the adequacy of diets to meet lysine needs of older adults can be assessed. The trial was registered at clinicaltrials.gov as NCT02008955 (https://clinicaltrials.gov/study/NCT02008955).

Iron overload induces dysplastic erythropoiesis and features of myelodysplasia in Nrf2-deficient mice.

Iron overload (IOL) is hypothesized to contribute to dysplastic erythropoiesis. Several conditions, including myelodysplastic syndrome, thalassemia and sickle cell anemia, are characterized by ineffective erythropoiesis and IOL. Iron is pro-oxidant and may participate in the pathophysiology of these conditions by increasing genomic instability and altering the microenvironment. There is, however, lack of in vivo evidence demonstrating a role of IOL and oxidative damage in dysplastic erythropoiesis. NRF2 transcription factor is the master regulator of antioxidant defenses, playing a crucial role in the cellular response to IOL in the liver. Here, we crossed Nrf2-/- with hemochromatosis (Hfe-/-) or hepcidin-null (Hamp1-/-) mice. Double-knockout mice developed features of ineffective erythropoiesis and myelodysplasia including macrocytic anemia, splenomegaly, and accumulation of immature dysplastic bone marrow (BM) cells. BM cells from Nrf2/Hamp1-/- mice showed increased in vitro clonogenic potential and, upon serial transplantation, recipients disclosed cytopenias, despite normal engraftment, suggesting defective differentiation. Unstimulated karyotype analysis showed increased chromosome instability and aneuploidy in Nrf2/Hamp1-/- BM cells. In HFE-related hemochromatosis patients, NRF2 promoter SNP rs35652124 genotype TT (predicted to decrease NRF2 expression) associated with increased MCV, consistent with erythroid dysplasia. Our results suggest that IOL induces ineffective erythropoiesis and dysplastic hematologic features through oxidative damage in Nrf2-deficient cells.

The Large-Scale Implementation of a Health Information System in Brazilian University Hospitals: Process and Outcomes.

Governments around the globe are paving the way for healthcare services that can have a profound impact on the overall well-being and development of their nations. However, government programs to implement health information technologies on a large-scale are challenging, especially in developing countries. In this article, the process and outcomes of the large-scale implementation of a hospital information system for the management of Brazilian university hospitals are analyzed. Based on a qualitative approach, this research involved 21 hospitals and comprised a documentary search, interviews with 24 hospital managers and two system user focus groups, and a questionnaire of 736 respondents. Generally, we observed that aspects relating to the wider context of system implementation (macro level), the managerial structure, cultural nuances, and political dynamics within each hospital (meso level), as well as the technology, work activities, and individuals themselves (micro level) acted as facilitators and/or obstacles to the implementation process. The dynamics and complex interactions established between these aspects had repercussions on the process, including the extended time necessary to implement the national program and the somewhat mixed outcomes obtained by hospitals in the national network. Mostly positive, these outcomes were linked to the eight emerging dimensions of practices and work processes; planning, control, and decision making; transparency and accountability; optimization in the use of resources; productivity of professionals; patient information security; safety and quality of care; and improvement in teaching and research. We argued here that to maximize the potential of information technology in healthcare on a large-scale, an integrative and cooperative vision is required, along with a high capacity for change management, considering the different regional, local, and institutional contexts.

Allogenic Synovia-Derived Mesenchymal Stem Cells for Treatment of Equine Tendinopathies and Desmopathies-Proof of Concept.

Tendon and ligament injuries are frequent in sport horses and humans, and such injuries represent a significant therapeutic challenge. Tissue regeneration and function recovery are the paramount goals of tendon and ligament lesion management. Nowadays, several regenerative treatments are being developed, based on the use of stem cell and stem cell-based therapies. In the present study, the preparation of equine synovial membrane mesenchymal stem cells (eSM-MSCs) is described for clinical use, collection, transport, isolation, differentiation, characterization, and application. These cells are fibroblast-like and grow in clusters. They retain osteogenic, chondrogenic, and adipogenic differentiation potential. We present 16 clinical cases of tendonitis and desmitis, treated with allogenic eSM-MSCs and autologous serum, and we also include their evaluation, treatment, and follow-up. The concerns associated with the use of autologous serum as a vehicle are related to a reduced immunogenic response after the administration of this therapeutic combination, as well as the pro-regenerative effects from the growth factors and immunoglobulins that are part of its constitution. Most of the cases (14/16) healed in 30 days and presented good outcomes. Treatment of tendon and ligament lesions with a mixture of eSM-MSCs and autologous serum appears to be a promising clinical option for this category of lesions in equine patients.

Repurposing Benztropine, Natamycin, and Nitazoxanide Using Drug Combination and Characterization of Gastric Cancer Cell Lines.

Gastric cancer (GC) ranked as the fifth most incident cancer in 2020 and the third leading cause of cancer mortality. Surgical prevention and radio/chemotherapy are the main approaches used in GC treatment, and there is an urgent need to explore and discover innovative and effective drugs to better treat this disease. A new strategy arises with the use of repurposed drugs. Drug repurposing coupled with drug combination schemes has been gaining interest in the scientific community. The main objective of this project was to evaluate the therapeutic effects of alternative drugs in GC. For that, three GC cell lines (AGS, MKN28, and MKN45) were used and characterized. Cell viability assays were performed with the reference drug 5-fluororacil (5-FU) and three repurposed drugs: natamycin, nitazoxanide, and benztropine. Nitazoxanide displayed the best results, being active in all GC cells. Further, 5-FU and nitazoxanide in combination were tested in MKN28 GC cells, and the results obtained showed that nitazoxanide alone was the most promising drug for GC therapy. This work demonstrated that the repurposing of drugs as single agents has the ability to decrease GC cell viability in a concentration-dependent manner.

Surgeon experience, robotic perioperative outcomes, and complications in gynecology.

OBJECTIVE: Robotic surgery is currently on the rise and has been widely applied all over the world. Gynecology offers great opportunities for the development of innovative techniques due to the magnitude of surgical needs. The aim of this study was to correlate perioperative complications, surgical time, and length of hospital stay with surgical diagnosis, procedure performed, and surgeon experience in robot-assisted gynecological surgeries in a 10-year period. METHODS: This was a retrospective, transversal, cross-sectional study involving 632 patients who underwent robotic gynecological surgery from January 2008 to December 2017 in a community hospital in Sao Paulo, Brazil. Medical records of robot-assisted gynecological operations were searched for perioperative complications, operative time, and length of hospital stay, correlating these outcomes with surgical diagnosis, procedure performed, and surgeon experience, considering those with 20 or less robotic procedures and surgeons with more than 20 cases in their career as in-training or qualified surgeons, respectively. RESULTS: Endometriosis (381 cases) was the most common surgical indication, followed by uterine myoma (171 patients). Qualified surgeons had 64% less complications than in-training surgeons (p=0.03) and achieved 20% lower surgical time and 15% shorter length of hospital stay. CONCLUSION: In this study, qualified surgeons with more than 20 robotic procedures had better perioperative outcomes and less complications than in-training surgeons during their first 20 robotic surgeries.

Can the Synergic Contribution of Multigenic Variants Explain the Clinical and Cellular Phenotypes of a Neurodevelopmental Disorder?

We describe an infant female with a syndromic neurodevelopmental clinical phenotype and increased chromosome instability as cellular phenotype. Genotype characterization revealed heterozygous variants in genes directly or indirectly linked to DNA repair: a de novo X-linked HDAC8 pathogenic variant, a paternally inherited FANCG pathogenic variant and a maternally inherited BRCA2 variant of uncertain significance. The full spectrum of the phenotype cannot be explained by any of the heterozygous variants on their own; thus, a synergic contribution is proposed. Complementation studies showed that the FANCG gene from the Fanconi Anaemia/BRCA (FA/BRCA) DNA repair pathway was impaired, indicating that the variant in FANCG contributes to the cellular phenotype. The patient's chromosome instability represents the first report where heterozygous variant(s) in the FA/BRCA pathway are implicated in the cellular phenotype. We propose that a multigenic contribution of heterozygous variants in HDAC8 and the FA/BRCA pathway might have a role in the phenotype of this neurodevelopmental disorder. The importance of these findings may have repercussion in the clinical management of other cases with a similar synergic contribution of heterozygous variants, allowing the establishment of new genotype-phenotype correlations and motivating the biochemical study of the underlying mechanisms.

Rat Olfactory Mucosa Mesenchymal Stem/Stromal Cells (OM-MSCs): A Characterization Study.

Stem/stromal cell-based therapies are a branch of regenerative medicine and stand as an attractive option to promote the repair of damaged or dysfunctional tissues and organs. Olfactory mucosa mesenchymal stem/stromal cells have been regarded as a promising tool in regenerative therapies because of their several favorable properties such as multipotency, high proliferation rate, helpful location, and few associated ethical issues. These cells are easily accessible in the nasal cavity of most mammals, including the rat, can be easily applied in autologous treatments, and do not cope with most of the obstacles associated with the use of other stem cells. Despite this, its application in preclinical trials and in both human and animal patients is still limited because of the small number of studies performed so far and to the nonexistence of a standard and unambiguous protocol for collection, isolation, and therapeutic application. In the present work a validation of a protocol for isolation, culture, expansion, freezing, and thawing of olfactory mucosa mesenchymal stem/stromal cells was performed, applied to the rat model, as well as a biological characterization of these cells. To investigate the therapeutic potential of OM-MSCs and their eventual safe application in preclinical trials, the main characteristics of OMSC stemness were addressed.

Mitoprotective Clinical Strategies in Type 2 Diabetes and Fanconi Anemia Patients: Suggestions for Clinical Management of Mitochondrial Dysfunction.

Oxidative stress (OS) and mitochondrial dysfunction (MDF) occur in a number of disorders, and several clinical studies have attempted to counteract OS and MDF by providing adjuvant treatments against disease progression. The present review is aimed at focusing on two apparently distant diseases, namely type 2 diabetes (T2D) and a rare genetic disease, Fanconi anemia (FA). The pathogenetic links between T2D and FA include the high T2D prevalence among FA patients and the recognized evidence for OS and MDF in both disorders. This latter phenotypic/pathogenetic feature-namely MDF-may be regarded as a mechanistic ground both accounting for the clinical outcomes in both diseases, and as a premise to clinical studies aimed at counteracting MDF. In the case for T2D, the working hypothesis is raised of evaluating any in vivo decrease of mitochondrial cofactors, or mitochondrial nutrients (MNs) such as α-lipoic acid, coenzyme Q10, and l-carnitine, with possibly combined MN-based treatments. As for FA, the established knowledge of MDF, as yet only obtained from in vitro or molecular studies, prompts the requirement to ascertain in vivo MDF, and to design clinical studies aimed at utilizing MNs toward mitigating or delaying FA's clinical progression. Altogether, this paper may contribute to building hypotheses for clinical studies in a number of OS/MDF-related diseases.

Safety model for the introduction of robotic surgery in gynecology / Modelo de segurança para a introdução da cirurgia robótica em ginecologia

Abstract Objective To analyze the perioperative results and safety of performing gynecological surgeries using robot-assisted laparoscopy during implementation of the technique in a community hospital over a 6-year period. Methods This was a retrospective observational study in which the medical records of 274 patients who underwent robotic surgery from September 2008 to December 2014 were analyzed. We evaluated age, body mass index (BMI), diagnosis, procedures performed, American Society of Anesthesiologists (ASA) classification, the presence of a proctor (experienced surgeon with at least 20 robotic cases), operative time, transfusion rate, perioperative complications, conversion rate, length of stay, referral to the intensive care unit (ICU), and mortality. We compared transfusion rate, perioperative complications and conversion rate between procedures performed by experienced and beginner robotic surgeons assisted by an experienced proctor. Results During the observed period, 3 experienced robotic surgeons performed 187 surgeries,while 87 surgeries were performedby 20 less experienced teams, always with the assistance of a proctor. The median patient age was 38 years, and the median BMI was 23.3 kg/m2. The most frequent diagnosis was endometriosis (57%) and the great majority of the patients were classified as ASA I or ASA II (99.6%). The median operative time was 225 minutes, and the median length of stay was 2 days. We observed a 5.8% transfusion rate, 0.8% rate of perioperative complications, 1.1% conversion rate to laparoscopy or laparotomy, no patients referred to ICU, and no deaths. There were no differences in transfusion, complications and conversion rates between experienced robotic surgeons and beginner robotic surgeons assisted by an experienced proctor. Conclusion In our casuistic, robot-assisted laparoscopy demonstrated to be a safe technique for gynecological surgeries, and the presence of an experienced proctor was considered a highlight in the safety model adopted for the introduction of the robotic gynecological surgery in a high-volume hospital and, mainly, for its extension among several surgical teams, assuring patient safety.

Resumo Objetivo Analisar os resultados perioperatórios e a segurança da realização de cirurgias ginecológicas por laparoscopia robô-assistida durante a implementação da técnica num hospital comunitário ao longo de 6 anos. Métodos Este foi umestudo retrospectivo observacional, comanálise dos prontuários de 274 pacientes que se submeteramà cirurgia robótica de setembro de 2008 a dezembro de 2014. Avaliamos idade, índice de massa corpórea (IMC), diagnóstico, procedimentos realizados, classificação da Sociedade Americana de Anestesiologia (ASA), presença de um preceptor (cirurgião experiente, compelomenos 20casos robóticos), tempocirúrgico, taxa de transfusão, complicações perioperatórias, taxa de conversão, tempo de internação, encaminhamento para Unidade de Terapia Intensiva (UTI) e mortalidade. Comparamos taxa de transfusão, complicações perioperatórias e taxa de conversão entre procedimentos realizados por cirurgiões experientes com a técnica e cirurgiões iniciantes na robótica, sempre assistidos por um preceptor experiente. Resultados Durante o período observado, 3 cirurgiões experientes realizaram 187 cirurgias, enquanto que 87 cirurgias foram realizadas por 20 equipes menos experientes, sempre com a presença de um preceptor. A mediana da idade foi 38 anos, e a mediana do IMC foi 23,3 kg/m2. O diagnósticomais frequente foi endometriose (57%) e a grande maioria das pacientes foi classificada como ASA I ou ASA II (99,6%). O tempo de cirurgia teve uma mediana de 225 minutos, e o tempo de permanência hospitalar teve uma mediana de 2 dias. Observamos 5,8% de taxa de transfusão, 0,8% de taxa de complicações perioperatórias, 1,1% de taxa de conversão para laparoscopia ou laparotomia e não houve pacientes encaminhadas à UTI, nem óbitos. Não houve diferença nos índices de transfusão, complicações e conversão entre cirurgiões experientes e cirurgiões iniciantes na robótica, assistidos por umpreceptor experiente. Conclusão Em nossa casuística, a laparoscopia robô-assistida demonstrou ser uma técnica segura para cirurgias ginecológicas, e a presença de um preceptor experiente foi considerada um ponto de destaque no modelo de segurança adotado para a introdução da cirurgia robótica em ginecologia num hospital de grande volume e, principalmente, na sua expansão entre diversas equipes cirúrgicas, mantendo a segurança das pacientes.

Safety Model for the Introduction of Robotic Surgery in Gynecology.

OBJECTIVE: To analyze the perioperative results and safety of performing gynecological surgeries using robot-assisted laparoscopy during implementation of the technique in a community hospital over a 6-year period. METHODS: This was a retrospective observational study in which the medical records of 274 patients who underwent robotic surgery from September 2008 to December 2014 were analyzed. We evaluated age, body mass index (BMI), diagnosis, procedures performed, American Society of Anesthesiologists (ASA) classification, the presence of a proctor (experienced surgeon with at least 20 robotic cases), operative time, transfusion rate, perioperative complications, conversion rate, length of stay, referral to the intensive care unit (ICU), and mortality. We compared transfusion rate, perioperative complications and conversion rate between procedures performed by experienced and beginner robotic surgeons assisted by an experienced proctor. RESULTS: During the observed period, 3 experienced robotic surgeons performed 187 surgeries, while 87 surgeries were performed by 20 less experienced teams, always with the assistance of a proctor. The median patient age was 38 years, and the median BMI was 23.3 kg/m2. The most frequent diagnosis was endometriosis (57%) and the great majority of the patients were classified as ASA I or ASA II (99.6%). The median operative time was 225 minutes, and the median length of stay was 2 days. We observed a 5.8% transfusion rate, 0.8% rate of perioperative complications, 1.1% conversion rate to laparoscopy or laparotomy, no patients referred to ICU, and no deaths. There were no differences in transfusion, complications and conversion rates between experienced robotic surgeons and beginner robotic surgeons assisted by an experienced proctor. CONCLUSION: In our casuistic, robot-assisted laparoscopy demonstrated to be a safe technique for gynecological surgeries, and the presence of an experienced proctor was considered a highlight in the safety model adopted for the introduction of the robotic gynecological surgery in a high-volume hospital and, mainly, for its extension among several surgical teams, assuring patient safety.

OBJETIVO: Analisar os resultados perioperatórios e a segurança da realização de cirurgias ginecológicas por laparoscopia robô-assistida durante a implementação da técnica num hospital comunitário ao longo de 6 anos. MéTODOS: Este foi um estudo retrospectivo observacional, com análise dos prontuários de 274 pacientes que se submeteram à cirurgia robótica de setembro de 2008 a dezembro de 2014. Avaliamos idade, índice de massa corpórea (IMC), diagnóstico, procedimentos realizados, classificação da Sociedade Americana de Anestesiologia (ASA), presença de um preceptor (cirurgião experiente, com pelo menos 20 casos robóticos), tempo cirúrgico, taxa de transfusão, complicações perioperatórias, taxa de conversão, tempo de internação, encaminhamento para Unidade de Terapia Intensiva (UTI) e mortalidade. Comparamos taxa de transfusão, complicações perioperatórias e taxa de conversão entre procedimentos realizados por cirurgiões experientes com a técnica e cirurgiões iniciantes na robótica, sempre assistidos por um preceptor experiente. RESULTADOS: Durante o período observado, 3 cirurgiões experientes realizaram 187 cirurgias, enquanto que 87 cirurgias foram realizadas por 20 equipes menos experientes, sempre com a presença de um preceptor. A mediana da idade foi 38 anos, e a mediana do IMC foi 23,3 kg/m2. O diagnóstico mais frequente foi endometriose (57%) e a grande maioria das pacientes foi classificada como ASA I ou ASA II (99,6%). O tempo de cirurgia teve uma mediana de 225 minutos, e o tempo de permanência hospitalar teve uma mediana de 2 dias. Observamos 5,8% de taxa de transfusão, 0,8% de taxa de complicações perioperatórias, 1,1% de taxa de conversão para laparoscopia ou laparotomia e não houve pacientes encaminhadas à UTI, nem óbitos. Não houve diferença nos índices de transfusão, complicações e conversão entre cirurgiões experientes e cirurgiões iniciantes na robótica, assistidos por um preceptor experiente. CONCLUSãO: Em nossa casuística, a laparoscopia robô-assistida demonstrou ser uma técnica segura para cirurgias ginecológicas, e a presença de um preceptor experiente foi considerada um ponto de destaque no modelo de segurança adotado para a introdução da cirurgia robótica em ginecologia num hospital de grande volume e, principalmente, na sua expansão entre diversas equipes cirúrgicas, mantendo a segurança das pacientes.

Caracterização de variáveis clínicas e do desenvolvimento motor de recém-nascidos prematuros / Characterization of clinical variables and neuromotor development of preterm infants

A Organização Mundial de Saúde considera prematuro aquele nascido com idade gestacional entre 20 a 37 semanas. Com os avanços tecnológicos existentes nas Unidades de Terapia Intensiva Neonatal, tem-se maior sobrevida destes, que apresentam desenvolvimento neuromotor inferior aos recém-nascidos a termo. Este estudo objetivou caracterizar o desenvolvimento motor e variáveis clínicas de prematuros nascidos em um hospital público. Trata- se de um estudo misto. A pontuação na Escala Motora Infantil de Alberta, peso ao nascimento, idade gestacional, tempo de internação na Unidade de Terapia Intensiva Neonatal e Apgar de 1o e 5o minuto de prematuros com idade corrigida entre 0 e 18 meses foram coletadas dos prontuários, entre maio de 2013 a abril de 2014. Para análise estatística, utilizou-se o teste do Qui-Quadrado (nível de significância p<0,05). Foram incluídos 267 prematuros, com mediana de idade gestacional de 32(30 - 33) semanas, de tempo de internação de 35 (25 - 58,5) dias e de peso médio de 1430 (11-75 - 1690) gramas. Foi encontrado em 10,49% das crianças o Apgar no 5o minuto inferior a 7. Verificou-se que 36,89% dos prematuros entre 0 e 5 meses, 39,6% entre 06 e 12 meses e 23,5% entre 13 e 18 meses estavam com riscos ou atrasos motores evidentes. Não foi observada correlação entre o Apgar do 5o minuto e o desenvolvimento motor. A maioria das crianças entre 06 e 12 meses apresentou risco para atrasos motores, justificando a importância do seguimento de prematuros em serviços de referência, mesmo após o primeiro ano de vida.

The World Health Organization considers preterm those born with a gestational age between 20 and 37 weeks. With the technological advances in Neonatal Intensive Care Units, we have seen a higher survival rate among preterms, who present lower neuromotor development than full term newborns. The aim of this study was to characterize the motor development and clinical variables of preterm infants born in a public hospital. It is a mixed study. The score on the Alberta Infant Motor Scale, birth weight, gestational age, time of stay in the Neonatal Intensive Care Unit, and the 1st and 5th minute Apgar of preterm infants with ages between 0 and 18 months corrected age, were collected from their files between May 2013 and April 2014. For statistical analysis, the Chisquare test (significance level of p < 0.05) was used. A total of 267 premature babies were included, with median gestational age of 32 (30-33) weeks, duration of stay of 35 (25-58.5) days, and mean weight of 1,430 (1,175-1,690) grams. A 5th minute Apgar score below 7 was found in 10.49% of the children. It was found that 36.89% of the infants between 0 and 5 months, 39.6% between 6 and 12 months, and 23.5% between 13 and 18 months of age were at risk of or had evident motor delay. No correlation was observed between the 5th minute Apgar and motor development. Most of the infants between 6 and 12 months were at risk for motor delay, justifying the importance of the follow-up for premature infants in specialized services even after the first year of life.

Increased red cell distribution width in Fanconi anemia: a novel marker of stress erythropoiesis.

BACKGROUND: Red cell distribution width (RDW), a classical parameter used in the differential diagnosis of anemia, has recently been recognized as a marker of chronic inflammation and high levels of oxidative stress (OS). Fanconi anemia (FA) is a genetic disorder associated to redox imbalance and dysfunctional response to OS. Clinically, it is characterized by progressive bone marrow failure, which remains the primary cause of morbidity and mortality. Macrocytosis and increased fetal hemoglobin, two indicators of bone marrow stress erythropoiesis, are generally the first hematological manifestations to appear in FA. However, the significance of RDW and its possible relation to stress erythropoiesis have never been explored in FA. In the present study we analyzed routine complete blood counts from 34 FA patients and evaluated RDW, correlating with the hematological parameters most consistently associated with the FA phenotype. RESULTS: We showed, for the first time, that RDW is significantly increased in FA. We also showed that increased RDW is correlated with thrombocytopenia, neutropenia and, most importantly, highly correlated with anemia. Analyzing sequential hemograms from 3 FA patients with different clinical outcomes, during 10 years follow-up, we confirmed a consistent association between increased RDW and decreased hemoglobin, which supports the postulated importance of RDW in the evaluation of hematological disease progression. CONCLUSIONS: This study shows, for the first time, that RDW is significantly increased in FA, and this increment is correlated with neutropenia, thrombocytopenia, and highly correlated with anemia. According to the present results, it is suggested that increased RDW can be a novel marker of stress erythropoiesis in FA.

Epigenetic Alterations in Fanconi Anaemia: Role in Pathophysiology and Therapeutic Potential.

Fanconi anaemia (FA) is an inherited disorder characterized by chromosomal instability. The phenotype is variable, which raises the possibility that it may be affected by other factors, such as epigenetic modifications. These play an important role in oncogenesis and may be pharmacologically manipulated. Our aim was to explore whether the epigenetic profiles in FA differ from non-FA individuals and whether these could be manipulated to alter the disease phenotype. We compared expression of epigenetic genes and DNA methylation profile of tumour suppressor genes between FA and normal samples. FA samples exhibited decreased expression levels of genes involved in epigenetic regulation and hypomethylation in the promoter regions of tumour suppressor genes. Treatment of FA cells with histone deacetylase inhibitor Vorinostat increased the expression of DNM3Tß and reduced the levels of CIITA and HDAC9, PAK1, USP16, all involved in different aspects of epigenetic and immune regulation. Given the ability of Vorinostat to modulate epigenetic genes in FA patients, we investigated its functional effects on the FA phenotype. This was assessed by incubating FA cells with Vorinostat and quantifying chromosomal breaks induced by DNA cross-linking agents. Treatment of FA cells with Vorinostat resulted in a significant reduction of aberrant cells (81% on average). Our results suggest that epigenetic mechanisms may play a role in oncogenesis in FA. Epigenetic agents may be helpful in improving the phenotype of FA patients, potentially reducing tumour incidence in this population.

[Histological classification and quality of life in women with endometriosis]. / Classificação histológica e qualidade de vida em mulheres portadoras de endometriose.

PURPOSE: To assess the relationship between the histological classification and the quality of life of patients operated for endometriosis. METHODS: A cross-sectional observational study, with assessment of 32 biopsies of the intestine, peritoneum and uterosacral ligament from 40 women with deep endometriosis. The quality of life (QOL) was determined by applying the SF-36 questionnaire pre-operatively and at 6 and 12 months postoperatively. Biopsies were histologically classified into pure stromal (EP), glandular differentiated (GD), glandular undifferentiated (GI) and mixed (GM), remaining in the sample only GI and GM, which are related to eight domains of the SF-36. RESULTS: According to the histologic type, the following distribution was observed: peritoneum 63% GI and 35% GM; intestine 19% GI and 24% GM; uterosacral ligament with 41% GI and 35% GM. Regarding the QOL and the histological classification, in the intestine only GM was associated with improvement of social and emotional aspects from 0 to 6 months; the domains general health status (p=0.01) and social aspect (p=0.04) were significantly related to improvement of the QOL from 0 to 6 months, and the general health status tended to improve from 0 to 12 months. Regarding pain (p=0.06) and the emotional aspect (p=0.05), the QOL tended to improve from 0 to 6 months and the vital capacity (p=0.1) improved from 0 to 6 months and from 0 to 12 months. Regarding the emotional aspect, evolution of the two histological types was not favorable for improvement in MG from 0 to 6 months. No significant relationships between histologic type and QOL were evident in the uterosacral ligament samples. CONCLUSION: Improvement in the QOL of women undergoing laparoscopic surgery for deep endometriosis is associated with the histologic grade. The peritoneal biopsy of GI revealed improved QOL after surgery.

Polyacrylic acid coated and non-coated iron oxide nanoparticles are not genotoxic to human T lymphocytes.

The use of iron oxide nanoparticles (ION) for diagnostic and therapeutic purposes requires a clear favorable risk-benefit ratio. This work was performed with the aim of studying the ability of polyacrylic acid (PAA)-coated and non-coated ION to induce genotoxicity in human T lymphocytes. For that purpose, their influence on cell cycle progression and on the induction of chromosome aberrations was evaluated. Blood samples collected from healthy human donors were exposed to PAA-coated and non-coated ION, at different concentrations, for 48h. The obtained results showed that, for all culture conditions, the tested ION are not genotoxic and do not influence the cell cycle arrest. Their possible cumulative effect with the iron-dependent genotoxic agent BLM was also evaluated. Blood samples collected from healthy human donors were exposed to ION, at different concentrations, for 48h, in the presence of a pre-determined toxic concentration of BLM. The obtained results showed that, for all culture conditions, the tested ION do not potentiate the clastogenic effects of BLM.

Increased levels of chromosomal aberrations and DNA damage in a group of workers exposed to formaldehyde.

Formaldehyde (FA) is a commonly used chemical in anatomy and pathology laboratories as a tissue preservative and fixative. Because of its sensitising properties, irritating effects and cancer implication, FA accounts probably for the most important chemical-exposure hazard concerning this professional group. Evidence for genotoxic effects and carcinogenic properties in humans is insufficient and conflicting, particularly in regard to the ability of inhaled FA to induce toxicity on other cells besides first contact tissues, such as buccal and nasal cells. To evaluate the effects of exposure to FA in human peripheral blood lymphocytes, a group of 84 anatomy pathology laboratory workers exposed occupationally to FA and 87 control subjects were tested for chromosomal aberrations (CAs) and DNA damage (comet assay). The level of exposure to FA in the workplace air was evaluated. The association between genotoxicity biomarkers and polymorphic genes of xenobiotic-metabolising and DNA repair enzymes were also assessed. The estimated mean level of FA exposure was 0.38±0.03 ppm. All cytogenetic endpoints assessed by CAs test and comet assay % tail DNA (%TDNA) were significantly higher in FA-exposed workers compared with controls. Regarding the effect of susceptibility biomarkers, results suggest that polymorphisms in CYP2E1 and GSTP1 metabolic genes, as well as, XRCC1 and PARP1 polymorphic genes involved in DNA repair pathways are associated with higher genetic damage in FA-exposed subjects. Data obtained in this study show a potential health risk situation of anatomy pathology laboratory workers exposed to FA (0.38 ppm). Implementation of security and hygiene measures may be crucial to decrease risk. The obtained information may also provide new important data to be used by health care programs and by governmental agencies responsible for occupational health and safety.

Classificação histológica e qualidade de vida em mulheres portadoras de endometriose / Histological classification and quality of life in women with endometriosis

OBJETIVO: Avaliar a relação entre a classificação histológica em pacientes operadas de endometriose e a qualidade de vida. MÉTODOS: Estudo observacional transversal, com avaliação de 32 biópsias de intestino, peritônio e ligamento uterossacro em 40 mulheres portadoras de endometriose profunda. Para a análise da qualidade de vida (QV) foi aplicado o questionário SF-36 antes da cirurgia e após 6 e 12 meses. As biópsias foram classificadas histologicamente em estromal puro (EP), glandular diferenciada (GD), indiferenciada (GI) e mista (GM), sendo que ficaram na amostra apenas o GI e GM, relacionadas aos oito domínios do SF-36. RESULTADOS: Observamos a seguinte distribuição de acordo com o tipo histológico: no peritônio 63% GI e 22% GM; no intestino 19% GI e 24% G e no uterossacro 41% GI e 35% GM. Quanto à QV e à classificação histológica, a evolução dos casos com implante no intestino mostrou que apenas o tipo GM apresentou melhora de 0 a 6 meses no aspecto social e no aspecto emocional. Ainda entre esses casos, o GI mostrou que os domínios estado geral da saúde (p=0,01) e aspecto social (p=0,04) têm relação significativa com a melhora da QV de 0 a 6 meses e uma tendência à melhora do estado geral da saúde de 0 a 12 meses. Quanto à dor (p=0,06) e ao aspecto emocional (p=0,05), observamos tendência à melhora na QV de 0 a 6 meses e na capacidade vital (p=0,1) de 0 a 6 e de 0 a 12 meses. No que se refere ao aspecto emocional, foi observado que entre as pacientes com tipo histológico GI, diferente do GM, não houve evolução favorável de 0 a 6 meses. No uterossacro não observamos relações significativas entre tipo histológico e QV. CONCLUSÃO: A melhora da QV em mulheres submetidas ao tratamento cirúrgico laparoscópico de endometriose profunda apresenta associação com grau de diferenciação histológica. Apenas as pacientes com endometriose classificada como indiferenciada e com lesões no peritônio mostraram melhora da QV após a cirurgia. .

PURPOSE: To assess the relationship between the histological classification and the quality of life of patients operated for endometriosis. METHODS: A cross-sectional observational study, with assessment of 32 biopsies of the intestine, peritoneum and uterosacral ligament from 40 women with deep endometriosis. The quality of life (QOL) was determined by applying the SF-36 questionnaire pre-operatively and at 6 and 12 months postoperatively. Biopsies were histologically classified into pure stromal (EP), glandular differentiated (GD), glandular undifferentiated (GI) and mixed (GM), remaining in the sample only GI and GM, which are related to eight domains of the SF-36. RESULTS: According to the histologic type, the following distribution was observed: peritoneum 63% GI and 35% GM; intestine 19% GI and 24% GM; uterosacral ligament with 41% GI and 35% GM. Regarding the QOL and the histological classification, in the intestine only GM was associated with improvement of social and emotional aspects from 0 to 6 months; the domains general health status (p=0.01) and social aspect (p=0.04) were significantly related to improvement of the QOL from 0 to 6 months, and the general health status tended to improve from 0 to 12 months. Regarding pain (p=0.06) and the emotional aspect (p=0.05), the QOL tended to improve from 0 to 6 months and the vital capacity (p=0.1) improved from 0 to 6 months and from 0 to 12 months. Regarding the emotional aspect, evolution of the two histological types was not favorable for improvement in MG from 0 to 6 months. No significant relationships between histologic type and QOL were evident in the uterosacral ligament samples. CONCLUSION: Improvement in the QOL of women undergoing laparoscopic surgery for deep endometriosis is associated with the histologic grade. The peritoneal biopsy of GI revealed improved QOL after surgery. .

Long-term outcomes on quality of life in women submitted to laparoscopic treatment for bowel endometriosis.

STUDY OBJECTIVE: To evaluate the long-term effects of laparoscopic surgery on quality of life in women with bowel endometriosis. DESIGN: Observational prospective cohort study (Canadian Task Force classification II). SETTING: Central Hospital of Santa Casa, Sao Paulo, Brazil. PATIENTS: Forty-five patients answered a short-form, 36-item, quality-of-life questionnaire (SF-36) at 3 different times. INTERVENTIONS: Between June 2007 and September 2008, patients underwent laparoscopic surgery to treat deep infiltrative endometriosis, with colorectal resection. MEASUREMENTS AND MAIN RESULTS: Forty-five patients with bowel endometriosis were followed up from 2007 to 2012. Before surgery, all patients exhibited signs suggestive of bowel endometriosis at magnetic resonance imaging and transrectal ultrasound. The patients underwent laparoscopic surgery for resection of the endometriosis lesions, including colorectal resection. The patients completed the questionnaire before surgery (T0), at 12 (T12) and 48 (T48) months after surgery. The 8 items of the SF-36 questionnaire at the different time points of application were compared. For each domain attribute, a score of 0 to 100 was assigned, where 0 signified the worst quality of life, and 100 the best. Statistical analysis was performed using analysis of variance. If differences were detected, multiple comparisons were performed using the Tukey test. Analysis of each domain revealed improved quality of life when comparing the period before surgery with 12 and 48 months after surgery. There was a significant increase (p < .001) in the scores in all of the SF-36 domains when comparing T0 vs T12 and T0 vs T48, with higher average scores at T48 corresponding to the domains of physical functioning, role physical, and social functioning (scores of 85.56, 75.69, and 73.61, respectively). CONCLUSION: Laparoscopic treatment of bowel endometriosis improved the long-term quality of life of patients.

Bone marrow cell transcripts from Fanconi anaemia patients reveal in vivo alterations in mitochondrial, redox and DNA repair pathways.

Fanconi anaemia (FA) is a genetic cancer predisposition disorder associated with cytogenetic instability, bone marrow failure and a pleiotropic cellular phenotype, including low thresholds of responses to oxidative stress, cross-linking agents and selected cytokines. This study was aimed at defining the scope of abnormalities in gene expression using the publicly available FA Transcriptome Consortium (FTC) database (Gene Expression Omnibus, 2009 and publicly available as GSE16334). We evaluated the data set that included transcriptomal analyses on RNA obtained from low-density bone marrow cells (BMC) from 20 patients with FA and 11 healthy volunteers, by seeking to identify changes in expression of over 22,000 genes, including a set of genes involved in: (i) bioenergetic pathways; (ii) antioxidant activities; (iii) response to stress and metal-chelating proteins; (iv) inflammation-related cytokines and (v) DNA repair. Ontological analysis of genes expressed at magnitudes of 1.5-fold or greater demonstrated significant suppression of genes in the categories of (i) energy metabolism; (ii) antioxidant activities; and (iii) stress and chelating proteins. Enhanced expression was found for 16 of 26 genes encoding inflammatory cytokines. A set of 20 of 21 transcripts for DNA repair activities were down-regulated; four of these transcripts related to type II topoisomerase. The data provide evidence for alterations in gene regulation of bioenergetic activities, redox-related activities, stress and metal-chelating proteins, and of some selected DNA repair activities in patients with FA.