Parkinson's disease and multiple system atrophy patient iPSC-derived oligodendrocytes exhibit alpha-synuclein-induced changes in maturation and immune reactive properties.

SignificanceOur results demonstrate the existence of early cellular pathways and network alterations in oligodendrocytes in the alpha-synucleinopathies Parkinson's disease and multiple system atrophy. They further reveal the involvement of an immune component triggered by alpha-synuclein protein, as well as a connection between (epi)genetic changes and immune reactivity in multiple system atrophy. The knowledge generated in this study could be used to devise novel therapeutic approaches to treat synucleinopathies.

Characteristics of patients diagnosed with cervical cancer in Brazil: preliminary results of the prospective cohort EVITA study (EVA001/LACOG 0215).

OBJECTIVES: Cervical cancer is the fourth most common cancer in women worldwide. Epidemiological and quality of life (QoL) data in patients with cervical cancer from low- and middle-income countries are scarce. We aimed to describe sociodemographic and clinicopathological characteristics and quality of life of patients with cervical cancer at diagnosis in Brazil. METHODS: EVITA is a prospective cohort study of newly diagnosed patients with cervical cancer from May 2016 to December 2017, stages I-IVB, from 16 Brazilian sites representing the five Brazilian regions. At baseline, medical evaluation was performed and European Organization for Research and Treatment of Cancer (EORTC) QLQ-CX24/C30 questionnaires were administered. RESULTS: A total of 631 patients were included. Mean±SD age was 49.3±13.9 years; skin color was non-white in 65.3%, and 68.0% had ≤8 years of formal education. In total, 85.1% of patients had a Pap smear. The main reasons reported by patients for not having a Pap smear were: lack of interest (46.9%), shame or embarrassment (19.7%), lack of knowledge (19.7%), and difficulty with access (9.1%). Most patients were diagnosed with locally advanced or metastatic disease (FIGO clinical stage II-IV in 81.8%- stage II in 35.2%, stage III in 36.1%, and stage IV in 10.5%). Patients with clinical stage III-IV had worse physical functioning and role functioning. CONCLUSIONS: Cervical cancer in Brazil is usually diagnosed at an advanced stage. Most patients have low formal education and are unemployed. Lack of interest was identified as a main reason for not having a screening test, and limited access was reported as a reason by <10% of the patients. Awareness campaigns must be a governmental priority, specially focused on the needy population, along with wide access to treatment.

Higher severity and risk of in-hospital mortality for COVID-19 patients with cancer during the year 2020 in Brazil: A countrywide analysis of secondary data.

BACKGROUND: Coronavirus disease 2019 (COVID-19) and cancer are serious public health problems worldwide. However, little is known about the risk factors of in-hospital mortality among COVID-19 patients with and without cancer in Brazil. The objective of this study was to evaluate the risk factors of in-hospital mortality among COVID-19 patients with and without cancer and to compare mortality according to gender and topography during the year 2020 in Brazil. METHODS: This was a secondary data study of hospitalized adult patients with a diagnosis of COVID-19 by real-time polymerase chain reaction testing in Brazil. The data were collected from the Influenza Epidemiological Surveillance Information System. RESULTS: This study analyzed data from 322,817 patients. The prevalence of cancer in patients with COVID-19 was 2.3%. COVID-19 patients with neurological diseases and cancer had the most lethal comorbidities in both sexes. COVID-19 patients with cancer were more likely to be older (median age, 67 vs 62 years; P < .001), to have a longer hospital stay (13.1 vs 11.5 days; P < .001), to be admitted to the intensive care unit (45.3% vs 39.6%; P < .001), to receive more invasive mechanical ventilation (27.1% vs 21.9%), and to have a higher risk of death (adjusted odds ratio [aOR], 1.94; 95% confidence interval [CI], 1.83-2.06; P < .001) than those without cancer. Patients with hematological neoplasia (aOR, 2.85; 95% CI, 2.41-3.38; P < .001) had a higher risk of mortality than those with solid tumors (aOR, 1.83; 95% CI, 1.72-1.95; P < .001) in both sexes. CONCLUSIONS: Brazilian COVID-19 patients with cancer have higher disease severity and a higher risk of mortality than those without cancer.

Phase II study of the safety and efficacy of the anti-PD-1 antibody balstilimab in patients with recurrent and/or metastatic cervical cancer.

OBJECTIVE: This phase II clinical trial evaluated the safety and antitumor activity of balstilimab, an anti-PD-1 antibody, in patients with previously-treated, recurrent/metastatic cervical cancer. METHODS: Eligible patients were 18 years or older with recurrent and/or metastatic cervical cancer and who had relapsed after a prior platinum-based treatment regimen for advanced disease. Balstilimab was administered intravenously at 3 mg/kg once every two weeks, for up to 24 months. The primary endpoint was objective response rate (ORR, RECIST v1.1) as assessed by an independent review committee. RESULTS: At data cutoff, 161 women (median age, 53 years [range 25-81]) were enrolled and treated with balstilimab. Of these, 140 had measurable disease at baseline and one prior line of platinum-based therapy in the metastatic, persistent, or recurrent setting; these patients were included in the efficacy analyses. The ORR was 15% (95% CI, 10.0%-21.8%) and included 5 patients with a complete response and 16 with a partial response. The median duration of response was 15.4 months. In patients with PD-L1-positive tumors the ORR was 20%, however patients with PD-L1-negative tumors also responded to balstilimab (ORR, 7.9%). Responses were not restricted to tumors of squamous cell histology, and an ORR of 12.5% was seen in the subset of patients with cervical adenocarcinoma. The disease control rate was 49.3% (95% CI, 41.1%-57.5%). Immune-mediated enterocolitis (3.1%) and diarrhea (1.9%) were the most common grade 3 or higher treatment-related adverse events. CONCLUSION: Balstilimab demonstrated meaningful and durable clinical activity, with manageable safety, in patients with previously-treated, recurrent/metastatic cervical cancer.

Phase II trial of neoadjuvant chemotherapy followed by chemoradiation in locally advanced cervical cancer.

OBJECTIVE: Cervical cancer is a global public health challenge. Since 1999, platin based chemoradiation (CRT) is the standard treatment for those patients with locally advanced disease. However, this population still has a dismal prognosis and, alternatives approaches such as adjuvant chemotherapy are controversial, especially because of increased toxicity. Neoadjuvant chemotherapy (NACT) could be an option for more intensive treatment with manageable toxicity. METHODS: A phase II, prospective, non-randomized trial was conducted at a reference center in Recife, Brazil. Locally advanced cervical cancer patients (Ib2-IVa) were treated with neoadjuvant cisplatin 35mg/m2 and gemcitabine 1000mg/m2 D1 and D8, for 2cycles. Then, they received CRT (50.4Gy) with weekly cisplatin 40mg/m2 followed by brachytherapy. Response rate (RR) and toxicity were the primary endpoints. Progression-free survival (PFS) and overall survival (OS) were secondary endpoints. RESULTS: Between Sep/2013 and Oct/2015, 50 patients were initiated on NACT and CRT. RR was 81% at the end of treatment. Hematological and gastrointestinal toxicity were most common. Grade 3/4 toxicity was 20% during NACT and 44% during CRT. Late adverse events were present in 20% of patients. PFS at 1 and 3-years were 73.4% (IC 58.7-83.6) and 53.9% (IC 36.9-68.3), respectively; and, OS at 1 and 3-years were 93.9% (IC 82.4-98.0) and 71.3% (IC 53.3-83.3), respectively. CONCLUSION: In our hands NACT in locally advanced cervical cancer patients did not show a meaningful improvement in ORR. Nevertheless, we believe it should be further explored in prospective trials.

Neoadjuvant Chemotherapy Followed by Chemoradiation in Cervical Carcinoma: A Review.

OBJECTIVE: Cervical cancer is a public health problem. A large proportion of patients have locally advanced (LA) disease at presentation and for this group, neoadjuvant chemotherapy (NACT) has an undefined role in the era of chemoradiotherapy. In countries with restricted access to radiotherapy, NACT may precipitate patients' access to an effective treatment approach. We carried out a systematic review to evaluate available data about NACT followed by chemoradiation. METHODS: Studies evaluated the use of NACT followed by chemoradiotherapy. Search strategy was performed in MEDLINE, LILACS, and Cochrane Library from 2003 to 2013. Conference proceedings of American Society of Clinical Oncology annual meeting, International Journal of Gynecological Cancer Society biennial meeting, and European Society of Gynecological Oncology biennial meeting were also reviewed. Key words used were "cervical uterine cancer," "cervical uterine neoplasm," "neoadjuvant therapy," "neoadjuvant treatment," and "locally advanced." Data collected included author information, year of publication, study design, number of participants, eligibility criteria, treatment, response rate (RR), disease-free survival, overall survival, and toxicities. RESULTS: Initial searches retrieved 7670 references. There were 7 eligible trials. Only 2 studies were published and a total of 323 participants were recruited. Patients with LA disease were eligible in most of them. Neoadjuvant chemotherapy consisted of paclitaxel combined with a platinum compound. Compliance was more than 90% in published trials. Response rate to NACT ranged from 67.8% to 70%. Major toxicity was hematological. Survival was evaluated in different points in time. Overall survival was up to 93% in 2 years, and in a poor prognostic group, 81% of patients were alive after 22 months. However, we must interpret these results with caution because of data limitation. CONCLUSIONS: Our result raises the possibility of NACT as an alternative upfront treatment for these patients with a small risk of disease progression. In countries where radiotherapy network is limited, randomized clinical trials should clarify its role.

Prevalence of contagious and environmental mastitis-causing bacteria in bulk tank milk and its relationships with milking practices of dairy cattle herds in São Miguel Island (Azores).

This study aimed to assess the degree of contamination of bulk tank milk (BTM) by Staphylococcus spp. and coliform bacteria and to identify major milking practices that help perpetuate them in dairy cattle herds in São Miguel Island. In July 2014, BTM was sampled and a survey concerning local milking practices was conducted on 100 herds. Semi quantitative multiplex polymerase chain reaction detected coagulase-negative staphylococci, Escherichia coli, Staphylococcus aureus, and other coliform bacteria (Klebsiella oxytoca, Klebsiella pneumoniae, and Serratia marcescens) in 100, 75, 59, and 35 % of BTM, respectively. According to multivariable univariate models, on herds not using hot water for cleaning the milking machine and teat liners, there was at least 3.4 more odds (P < 0.01) to have S. aureus or coliform bacteria contamination in BTM. The likelihood of finding S. aureus in BTM was higher (P < 0.001) on herds without high hygiene during milking, when milking mastitic cows at the end, on abrupt cessation of milking at dry-off, and official milk control implementation. The glove use also favored (odds ratio (OR) 5.8; P < 0.01) the detection of coliform bacteria in BTM. Poor milking practices identified in this study should be avoided in order to decrease S. aureus and coliform bacteria contamination of BTM. Other factors associated with milk quality in São Miguel Island also should be further investigated.

The psychometrics and validity of the Junior Temperament and Character Inventory in Portuguese adolescents.

Personality traits related to persistence and self-regulation of long-term goals can predict academic performance as well or better than measures of intelligence. The 5-factor model has been suggested to outperform some other personality tests in predicting academic performance, but it has not been compared to Cloninger's psychobiological model for this purpose. The aims of this study were, first, to evaluate the psychometric properties of the Junior Temperament and Character Inventory (JTCI) in adolescents in Portugal, and second, to evaluate the comparative validity of age-appropriate versions of Cloninger's 7-factor psychobiological model, Costa and McCrae's five-factor NEO-Personality Inventory-Revised, and Cattell's 16-personality-factor inventory in predicting academic achievement. All dimensions of the Portuguese JTCI had moderate to strong internal consistency. The Cattell's sixteen-personality-factor and NEO inventories provided strong construct validity for the JTCI in students younger than 17 years and for the revised adult version (TCI-Revised) in those 17 years and older. High TCI Persistence predicted school grades regardless of age as much or more than intelligence. High TCI Harm Avoidance, high Self-Transcendence, and low TCI Novelty Seeking were additional predictors in students older than 17. The psychobiological model, as measured by the JTCI and TCI-Revised, performed as well or better than other measures of personality or intelligence in predicting academic achievement.

Exploratory analysis of radiomic as prognostic biomarkers in 18F-FDG PET/CT scan in uterine cervical cancer.

Objective: To evaluate the performance of 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET/CT) radiomic features to predict overall survival (OS) in patients with locally advanced uterine cervical carcinoma. Methods: Longitudinal and retrospective study that evaluated 50 patients with cervical epidermoid carcinoma (clinical stage IB2 to IVA according to FIGO). Segmentation of the 18F-FDG PET/CT tumors was performed using the LIFEx software, generating the radiomic features. We used the Mann-Whitney test to select radiomic features associated with the clinical outcome (death), excluding the features highly correlated with each other with Spearman correlation. Subsequently, ROC curves and a Kaplan-Meier analysis were performed. A p-value < 0.05 were considered significant. Results: The median follow-up was 23.5 months and longer than 24 months in all surviving patients. Independent predictors for OS were found-SUVpeak with an AUC of 0.74, sensitivity of 77.8%, and specificity of 72.7% (p = 0.006); and the textural feature gray-level run-length matrix GLRLM_LRLGE, with AUC of 0.74, sensitivity of 72.2%, and specificity of 81.8% (p = 0.005). When we used the derived cut-off points from these ROC curves (12.76 for SUVpeak and 0.001 for GLRLM_LRLGE) in a Kaplan-Meier analysis, we can see two different groups (one with an overall survival probability of approximately 90% and the other with 30%). These biomarkers are independent of FIGO staging. Conclusion: By radiomic 18F-FDG PET/CT data analysis, SUVpeak and GLRLM_LRLGE textural feature presented the best performance to predict OS in patients with cervical cancer undergoing chemo-radiotherapy and brachytherapy.

Meningeal carcinomatosis in breast cancer: prognostic factors and outcome.

Meningeal carcinomatosis (MC) occurs in up to 5% of breast cancer patients. Few studies have evaluated prognostic markers in breast cancer patients with MC. Our aim was to describe the treatment of breast cancer patients with MC, and identify prognostic factors related to survival. Sixty breast cancer patients that had a diagnosis of MC between January 2003 and December 2009 were included. The median age was 46 years (range 27-76). Most patients had invasive ductal carcinoma (78.3%) and high histological/nuclear grade (61.7/53.3%). Estrogen and progesterone receptors were positive in 51.7 and 43.3% of patients, respectively, and 15% were HER-2-positive. Symptoms at presentation were headache, cranial nerve dysfunction, seizures, and intracranial hypertension signals. Diagnosis was made by CSF cytology in 66.7% of cases and by MRI in 71.7%. Intrathecal (IT) chemotherapy was used in 68.3% of patients, and 21.6% received a new systemic treatment (chemo- or hormone therapy). Median survival was 3.3 months (range 0.03-90.4). There was no survival difference according to age, nuclear grade, hormonal and HER-2 status, CSF features, sites of metastasis, systemic and IT chemotherapy, or radiotherapy. However, histological grade and performance status had a significant impact on survival in the multivariate analysis. Only four papers have addressed prognostic factors in breast cancer patients with MC in the last two decades. The results of those reports are discussed here. High histological grade and poor performance status seem to impact survival of breast cancer patients with MC. Prospective studies are necessary to clarify the role of IT and systemic treatment in the treatment of those patients.

Pathologic complete response with neoadjuvant imatinib for locally advanced pelvic GIST.

There are few data that show pathologic complete response (pCR) to imatinib treatment in gastrointestinal stromal tumors (GISTs). We describe a case of a patient with pCR of a pelvic, locally advanced, high-risk GIST who was treated with neoadjuvant imatinib and ultimately underwent a conservative procedure. A 48-year-old male presented with a pelvic mass 10 cm in diameter. Biopsy revealed a gastrointestinal stromal tumor of rectal origin. Although it was considered initially resectable, an extensive procedure would have been necessary for complete resection. Treatment with imatinib was initiated, resulting in partial response. The patient remained on imatinib for over 15 months, maintaining stable disease. Radical prostatectomy with anal sphincter preservation was performed. Pathological report revealed no viable neoplastic cells. The use of imatinib was held for 6 months after the surgery. At a follow-up 15 months after surgery, the patient had no evidence of disease. Our report may help to guide future studies of neoadjuvant imatinib for large pelvic or rectum GISTs that are initially considered unresectable.

QTest: A new way to easily sample, store, and ship samples to perform Q fever PCR analysis on bulk tank milk.

Bulk tank milk (BTM) is an easy, inexpensive, and representative sample for detection of Coxiella burnetii infections (Q fever) in dairy herds using real-time PCR. Bulk tank milk PCR can be performed either for initial herd screening or for monitoring the effectiveness of preventive measures. However, one major limitation under field conditions is the need to deliver BTM samples in adequate condition (quickly, safely, and under refrigeration) to a qualified laboratory. In addition, sending non-inactivated biological samples via normal mail may be prohibited. We developed an innovative, easy, and accurate diagnostic tool (QTest) for Q fever to support veterinarians and farmers in overcoming these constraints. The farmer or veterinarian simply places some drops of BTM on a Whatman FTA Elute Micro Card (FTA card) and lets the card dry before mailing it to the laboratory. In a 2-step study, we tested the hypotheses that (1) BTM samples stored on FTA cards are stable over time and at different temperatures, and (2) PCR results obtained via FTA cards are consistent with those obtained from raw BTM samples. The stability of C. burnetii DNA in milk preserved on an FTA card was maintained for at least 29 d at room temperature or 37°C to mimic field conditions. In our field study, of the original 70 positive BTM samples (when tested on raw BTM just after sampling), 58 samples were positive (on either raw BTM or FTA card) by the time of the direct comparison study (10 to 14 d later). Of these 58 samples, 45 raw BTM samples still tested positive after aging, and 53 FTA card BTM samples tested positive, indicating that detection was higher using FTA cards (91.4%) than raw milk (77.6%). Therefore, with inactivation and shipping advantages, this technique facilitates an easier and more practical approach to diagnosis of Q fever at the herd level and would support Q fever control strategies, especially in countries lacking adequate and close laboratory facilities.

TNF-α and α-synuclein fibrils differently regulate human astrocyte immune reactivity and impair mitochondrial respiration.

Here, we examine the cellular changes triggered by tumor necrosis factor alpha (TNF-α) and different alpha-synuclein (αSYN) species in astrocytes derived from induced pluripotent stem cells. Human astrocytes treated with TNF-α display a strong reactive pro-inflammatory phenotype with upregulation of pro-inflammatory gene networks, activation of the nuclear factor κB (NF-κB) pathway, and release of pro-inflammatory cytokines, whereas those treated with high-molecular-weight αSYN fibrils acquire a reactive antigen (cross)-presenting phenotype with upregulation of major histocompatibility complex (MHC) genes and increased human leukocyte antigen (HLA) molecules at the cell surface. Surprisingly, the cell surface location of MHC proteins is abrogated by larger F110 fibrillar polymorphs, despite the upregulation of MHC genes. Interestingly, TNF-α and αSYN fibrils compete to drive the astrocyte immune reactive response. The astrocyte immune responses are accompanied by an impaired mitochondrial respiration, which is exacerbated in Parkinson's disease (PD) astrocytes. Our data provide evidence for astrocytic involvement in PD pathogenesis and reveal their complex immune reactive responses to exogenous stressors.

Generation of an induced pluripotent stem cell line (CSC-32) from a patient with Parkinson's disease carrying a heterozygous variation p.A53T in the SNCA gene.

Here, we describe the generation of an induced pluripotent stem cell (iPSC) line, from a male patient diagnosed with Parkinson's disease (PD). The patient carries a heterozygous variation p.A53T in the SNCA gene. Skin fibroblasts were reprogrammed using the non-integrating Sendai virus technology to deliver OCT3/4, SOX2, c-MYC and KLF4 factors. The generated iPSC line (CSC-32) preserved the mutation, displayed expression of common pluripotency markers, differentiated into derivatives of the three germ layers, and exhibited a normal karyotype. The clone CSC-32B is presented thereafter; it can be used to study the mechanisms underlying PD pathogenesis.

Canine Rabies in the Brazil-Bolivia Border Region from 2006 to 2014.

BACKGROUND: After 2006 the cross-border region between the state of Mato Grosso do Sul (Brazil) and the Germán Busch Province (Bolivia) became risk areas for canine rabies antigenic variant 1, previously unknown in the Brazilian territory. OBJECTIVES: To perform a descriptive analysis of canine rabies from 2006 to 2014, analyzing the database of the official rabies diagnostic laboratory of the State Agency of Animal and Plant Health Protection of Mato Grosso do Sul. METHODS: A descriptive analysis of canine rabies from 2006 to 2014 was performed using the database of the official rabies diagnostic laboratory of the State Agency of Animal and Plant Health Protection of Mato Grosso do Sul. Location, time and residence status of the animals were analyzed. Monthly frequencies were calculated as the ratio of the number of positive samples to the total of sent samples and were then statistically compared. FINDINGS: In the period, 539 samples of nervous system from dogs and cats were sent for rabies diagnosis, of which 37 (6.9%; CI95% 5.0-9.3) canine and no positive feline samples were found positive. Twenty-four (64.9%, CI95% 48.8-78.2) positive samples were from Bolivia and 13 (31.1%, CI95% 21.8-51.2) from Brazil. Most positive animals were owned. The years 2008 and 2009 showed the highest occurrence of canine rabies, with 18 cases recorded in 2008 and 6 in 2009 (17 in Bolivia and 7 in Brazil). Annual samples sent in Brazil presented a decreasing trend (R2 = 0.53) and, over the months, a higher concentration of samples was observed between May and August (R2 = 0.69). No annual or monthly trends were observed for Bolivian samples (R2 < 0.003). CONCLUSIONS: AgV1 canine rabies due to antigenic variant 1 is still considered an endemic disease in the Brazil-Bolivia border region, requiring an international One Health Approach to mitigate canine rabies in Latin America.

Cellular alterations identified in pluripotent stem cell-derived midbrain spheroids generated from a female patient with progressive external ophthalmoplegia and parkinsonism who carries a novel variation (p.Q811R) in the POLG1 gene.

Variations in the POLG1 gene encoding the catalytic subunit of the mitochondrial DNA polymerase gamma, have recently been associated with Parkinson's disease (PD), especially in patients diagnosed with progressive external ophthalmoplegia (PEO). However, the majority of the studies reporting this association mainly focused on the genetic identification of the variation in POLG1 in PD patient primary cells, and determination of mitochondrial DNA copy number, providing little information about the cellular alterations existing in patient brain cells, in particular dopaminergic neurons. Therefore, through the use of induced pluripotent stem cells (iPSCs), we assessed cellular alterations in novel p.Q811R POLG1 (POLG1Q811R) variant midbrain dopaminergic neuron-containing spheroids (MDNS) from a female patient who developed early-onset PD, and compared them to cultures derived from a healthy control of the same gender. Both POLG1 variant and control MDNS contained functional midbrain regionalized TH/FOXA2-positive dopaminergic neurons, capable of releasing dopamine. Western blot analysis identified the presence of high molecular weight oligomeric alpha-synuclein in POLG1Q811R MDNS compared to control cultures. In order to assess POLG1Q811R-related cellular alterations within the MDNS, we applied mass-spectrometry based quantitative proteomic analysis. In total, 6749 proteins were identified, with 61 significantly differentially expressed between POLG1Q811R and control samples. Pro- and anti-inflammatory signaling and pathways involved in energy metabolism were altered. Notably, increased glycolysis in POLG1Q811R MDNS was suggested by the increase in PFKM and LDHA levels and confirmed using functional analysis of glycolytic rate and oxygen consumption levels. Our results validate the use of iPSCs to assess cellular alterations in relation to PD pathogenesis, in a unique PD patient carrying a novel p.Q811R variation in POLG1, and identify several altered pathways that may be relevant to PD pathogenesis.

Generation of an induced pluripotent stem cell line (CSC-46) from a patient with Parkinson's disease carrying a novel p.R301C mutation in the GBA gene.

Mutations in the glucocerebrosidase (GBA) gene have been associated with the development of Parkinson's disease (PD). An induced pluripotent stem cell (iPSC) line was generated from a 60-year old patient diagnosed with PD and carrying a new mutation variant p.R301C in GBA. Using non-integrating Sendai virus-based technology, we utilized OCT3/4, SOX2, c-MYC and KLF4 transcription factors to reprogram skin fibroblasts into iPSCs. The generated iPSC line retained the mutation, displayed expression of common pluripotency markers, differentiated into the three germ layers, and exhibited normal karyotype. The iPSC line can be further used for studying PD pathogenesis.

PARK2 Mutation Causes Metabolic Disturbances and Impaired Survival of Human iPSC-Derived Neurons.

The protein parkin, encoded by the PARK2 gene, is vital for mitochondrial homeostasis, and although it has been implicated in Parkinson's disease (PD), the disease mechanisms remain unclear. We have applied mass spectrometry-based proteomics to investigate the effects of parkin dysfunction on the mitochondrial proteome in human isogenic induced pluripotent stem cell-derived neurons with and without PARK2 knockout (KO). The proteomic analysis quantified nearly 60% of all mitochondrial proteins, 119 of which were dysregulated in neurons with PARK2 KO. The protein changes indicated disturbances in oxidative stress defense, mitochondrial respiration and morphology, cell cycle control, and cell viability. Structural and functional analyses revealed an increase in mitochondrial area and the presence of elongated mitochondria as well as impaired glycolysis and lactate-supported respiration, leading to an impaired cell survival in PARK2 KO neurons. This adds valuable insight into the effect of parkin dysfunction in human neurons and provides knowledge of disease-related pathways that can potentially be targeted for therapeutic intervention.

Alkydic resin wastewaters treatment by fenton and photo-Fenton processes.

Advanced oxidation processes are an emerging option to treatment of the painting industry effluents. The aim of this study was to compare the effectiveness of the Fenton and photo-Fenton processes in chemical oxygen demand (COD), total organic carbon (TOC) and phenolic compounds removal from wastewaters generated during alkydic resins manufacture. The optimized treatment conditions are the following: pH 3.0, 15.15x10(-3)molL(-1) FeSO(4) and 0.30molL(-1) H(2)O(2) for a reaction time of 6h. photo-Fenton experiments were carried out in the presence of sunlight or artificial radiation and complementary additions of H(2)O(2) were made for all experiments. The best results were obtained with photo-Fenton process assisted with solar radiation, with reductions of 99.5 and 99.1% of COD and TOC levels, respectively. Fenton and photo-Fenton (with artificial irradiation) processes presented lower but not insignificant removals, within 60-80% reduction for both COD and TOC. In addition, an excellent removal (95%) of total phenols was obtained using photo-Fenton process assisted with artificial irradiation. This study demonstrated that the use of photo-Fenton process on alkydic resins wastewater treatment is very promising especially when solar light is used.

Reporting on methods to generate and purify rodent and human oligodendrocytes from different sources.

Oligodendrocytes are part of the glial cells located in the central nervous system, capable of providing trophic support to neurons and ensheathing their axons. These cells can become dysfunctional under pathologic condition. Rodent and human pluripotent stem cells are inexhaustible sources for producing oligodendrocytes that can be used for disease modeling and cell replacement therapy studies. They also offer many opportunities to model the contribution of oligodendrocytes in non-genetic disorders such as multiple system atrophy. In this method article, we provide robust and reproducible differentiation protocols to obtain oligodendrocyte progenitor cells and purify them using fluorescence activated cell sorting.