Can ozone be used as antimicrobial in the dairy industry? A systematic review.

Milk and dairy products are abundantly consumed in all cultures, but unprocessed products can harbor pathogenic microorganisms that can cause serious health risks for its consumers. To avoid this, it is necessary to process the products. Ozonation is a clean technique that has antimicrobial power due to its oxidation potential, reducing the microorganisms and limiting the production of enzymes, but the effectiveness of ozone treatment can be affected by the temperature, pH, additives, humidity, and the amount of organic matter around the cells. The goal of this systematic review was to analyze whether the use of ozone could improve the microbiological quality of dairy products and whether it could be used as an antimicrobial technique. Six databases (PubMed, Scielo, CAPES, Science Direct, Science Core Collection, and PLOS) were used in this research, with 2 independent reviewers selecting articles up to November 21, 2020, with experiments that used ozone as an antimicrobial in dairy products. A total of 731 articles were found, but only 9 were selected. The remainder were excluded according to the following criteria: was not related to the main theme; was a review; did not contain microbiological analysis; did not mention the concentration of gas and time of the ozone treatment; and was not an experiment. Important points were noted in quality criteria, which resulted in the need to standardize the methodology applied in research to improve the quality of the experiments. Studies were carried out with many different samples of milk, but the best results in reducing the microorganism count were obtained from samples containing low levels of fat.

Genetic profile of Brazilian patients with dystrophinopathies.

Different types of mutations in the DMD gene underlie Duchenne muscular dystrophies (DMD) and Becker muscular dystrophies (BMD). Large deletions and duplications are the most frequent causative genetic alterations worldwide, but little is known about DMD/BMD genetic profile in Brazil. Hence, we recruited patients with DMD and BMD from 8 neuromuscular reference centers along the country, and performed a comprehensive molecular investigation that included Multiplex Ligation-dependent Probe Amplification and Next generation sequencing (NGS) analyses. We evaluated 199 patients from 177 unrelated families: 166 with DMD, 32 with BMD and 1 1.5 years old asymptomatic patient with persistent hiperCKemia. Overall, large deletions (58.2%) followed by nonsense mutations (12.4%) and large duplications (11.3%) were the most frequent variants in Brazilian families. Large deletions were less frequent in BMD than in DMD (44.8% vs 60.8%). We identified 19 new DMD variants. Nonsense mutations were significantly more frequent in patients from northeastern region than from southern/southeastern regions of Brazil (27.7% vs 8.5%, P < .05). Genetic profile of Brazilian patients with DMD/BMD is similar to previously reported cohorts, but it is not uniform across the country. This information is important to plan rational clinical care for patients in face of the new coming mutation-specific therapies.

Control of Immune Response to Allogeneic Embryonic Stem Cells by CD3 Antibody-Mediated Operational Tolerance Induction.

Implantation of embryonic stem cells (ESCs) and their differentiated derivatives into allogeneic hosts triggers an immune response that represents a hurdle to clinical application. We established in autoimmunity and in transplantation that CD3 antibody therapy induces a state of immune tolerance. Promising results have been obtained with CD3 antibodies in the clinic. In this study, we tested whether this strategy can prolong the survival of undifferentiated ESCs and their differentiated derivatives in histoincompatible hosts. Recipients of either mouse ESC-derived embryoid bodies (EBs) or cardiac progenitors received a single short tolerogenic regimen of CD3 antibody. In immunocompetent mice, allogeneic EBs and cardiac progenitors were rejected within 20-25 days. Recipients treated with CD3 antibody showed long-term survival of implanted cardiac progenitors or EBs. In due course, EBs became teratomas, the growth of which was self-limited. Regulatory CD4(+)FoxP3(+) T cells and signaling through the PD1/PDL1 pathway played key roles in the CD3 antibody therapeutic effect. Gene profiling emphasized the importance of TGF-ß and the inhibitory T cell coreceptor Tim3 to the observed effect. These results demonstrate that CD3 antibody administered alone promotes prolonged survival of allogeneic ESC derivatives and thus could prove useful for enhancing cell engraftment in the absence of chronic immunosuppression.

Probing Single-Cell Macrophage Polarization and Heterogeneity Using Thermo-Reversible Hydrogels in Droplet-Based Microfluidics.

Human immune cells intrinsically exist as heterogenous populations. To understand cellular heterogeneity, both cell culture and analysis should be executed with single-cell resolution to eliminate juxtacrine and paracrine interactions, as these can lead to a homogenized cell response, obscuring unique cellular behavior. Droplet microfluidics has emerged as a potent tool to culture and stimulate single cells at high throughput. However, when studying adherent cells at single-cell level, it is imperative to provide a substrate for the cells to adhere to, as suspension culture conditions can negatively affect biological function and behavior. Therefore, we combined a droplet-based microfluidic platform with a thermo-reversible polyisocyanide (PIC) hydrogel, which allowed for robust droplet formation at low temperatures, whilst ensuring catalyzer-free droplet gelation and easy cell recovery after culture for downstream analysis. With this approach, we probed the heterogeneity of highly adherent human macrophages under both pro-inflammatory M1 and anti-inflammatory M2 polarization conditions. We showed that co-encapsulation of multiple cells enhanced cell polarization compared to single cells, indicating that cellular communication is a potent driver of macrophage polarization. Additionally, we highlight that culturing single macrophages in PIC hydrogel droplets displayed higher cell viability and enhanced M2 polarization compared to single macrophages cultured in suspension. Remarkably, combining phenotypical and functional analysis on single cultured macrophages revealed a subset of cells in a persistent M1 state, which were undetectable in conventional bulk cultures. Taken together, combining droplet-based microfluidics with hydrogels is a versatile and powerful tool to study the biological function of adherent cell types at single-cell resolution with high throughput.

Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection.

The World Health Organization (WHO) has declared a pandemic of COVID-19, the disease caused by the recently described SARS-CoV-2. The relevance and importance of mass diagnosis in order to find the asymptomatic individuals is widely recognized as a mandatory tool to reinforce the control measures for monitoring virus circulation and reduce the spreading of SARS-CoV-2. Here, we described quickness and cheaper strategies of direct RT-qPCR (in the absence of RNA isolation) and compared the results to those obtained using standard RNA isolation procedure. The tests varied using pure, diluted samples, combined with Proteinase K (PK) or Lysis Buffer. Our findings showed consistently that PK pre-treated samples in the absence of RNA extraction procedures presents similar results to those obtained by standard RNA isolation procedures. On average, 16 samples extracted with the MagMAX™ CORE Kit, take around 2 h, costing an average of USD 5, the pre-treatment of samples using PK, on the other hand, would cut the value to less than USD 0.30 and reduce the time of procedure in more than 1 ½ hours. The present study suggests the use of PK treatment instead of RNA isolation in order to reduce costs and time in processing samples for molecular diagnosis of SARS-CoV-2.

Clinical experiences of using a cellulose dressing on burns and donor site wounds.

These preliminary results suggest this temporary skin substitute reduces pain and achieves comparable healing rates to other dressings. Furthermore, its transparency enables regular inspection of the wound bed for signs of infection.

Development of compact UASB/trickling filter systems for treating domestic wastewater in small communities in Brazil.

The paper analyses the concept and performance of compact UASB/TF systems in relation to configuration, use of innovative packing media and operational conditions. For the conditions tested, the UASB/TF systems had consistently complied with the Brazilian discharge standard regarding to COD, BOD and TSS parameters. However, some enhancements are still necessary in order to increase nitrification in the process. The conditions to promote nitrification in shallow TF (packed bed up to 2.50 m height) seem to be compatible with the proposed simplification in the flowsheet, which is to operate the system without secondary clarifiers.

Diffuse embryoma of the testis. A distinctive form of mixed germ cell tumor.

Two testicular tumors characterized by a diffuse, orderly arrangement of embryonal, yolk sac, and trophoblastic elements are described as examples of a newly recognized form of mixed germ cell neoplasia. In one case, ribbons of embryonal carcinoma and yolk sac tumor wound around one another to create a distinctive necklace pattern. In the second case, differentiation of the yolk sac component was more advanced with the formation of numerous clusters of cells resembling hepatocytes. Tumors with these patterns are appropriately designated diffuse embryomas to distinguish them from polyembryomas and other forms of malignant mixed germ cell tumor.

Characterization of immature sinus histiocytes (monocytoid cells) in reactive lymph nodes by use of monoclonal antibodies.

The distinctive mononuclear cells that appear in the cortical sinuses of lymph nodes in toxoplasma lymphadenitis and other conditions have been termed "immature sinus histiocytes," although these cells have neither enzyme histochemical nor ultrastructural features of histiocytes. The authors stained these cells in frozen sections of six reactive lymph nodes, using the immunoperoxidase technique, with monoclonal antibodies to T cell, B cell, and monocyte antigens and with heteroantisera to immunoglobulin and lysozyme. The immature sinus histiocytes stained with a monoclonal antibody to B cells (anti-B1) and had immunoglobulin of the IgG class. They did not react with monoclonal antibodies to T cells or monocytes or with antilysozyme. In contrast, medullary sinus histiocytes in three additional lymph nodes were B1-negative and reacted with anti-T4, anti-M1, and anti-lysozyme. These results indicate that immature sinus histiocytes are IgG-bearing B lymphocytes rather than histiocytes. Their role in the immune response remains enigmatic.

Cystic teratomas of the pancreas.

Cystic teratomas of the pancreas constitute an extremely rare entity with only nine cases, to our knowledge, described in the world literature. Symptoms are usually due to the compressive effects of the tumor on the neighboring organs. They should be considered in the differential diagnosis of slow-growing benign pancreatic cysts. We describe a 25-year-old woman with a pancreatic teratoma who was operated on in 1976 with the diagnosis of calcified pancreatic cyst. The diagnostic and surgical procedures are described, as well as a 14-year follow-up. The previously published cases are reviewed and the differential diagnosis is discussed. Early diagnosis and the need for total tumor resection are emphasized.

Diagnostic evaluation of PCR in goats experimentally infected with Trypanosoma vivax.

Six goats were experimentally infected with a stock of Trypanosoma vivax. Parasitaemia was weekly monitored by buffy coat and wet blood film examination during a period of 15 weeks and another 3 weeks following drug-treatment. Dried blood samples were tested by the polymerase chain reaction (PCR), using an extraction method with Chelex 100 (BioRad). PCR proved consistently more sensitive than the parasitological techniques.

Diagnostic evaluation of PCR on dried blood samples from goats experimentally infected with Trypanosoma brucei brucei.

In seven goats experimentally infected with a pleomorphic clone of Trypanosoma brucei brucei, parasitaemia was monitored weekly for 6 weeks by wet blood film and microhaematocrit buffy coat examination. Dried blood samples on filter paper were concomitantly collected and tested by PCR using three different primer sets, putatively specific for Trypanozoon, T. vivax and T. congolense. With the originally designed ORPHON5J Trypanozoon primers, PCR tests became positive after 1 week (six animals) or 2 weeks (one animal) of infection and remained consistently positive until the end of the experiments, thus yielding an overall positivity rate of 97%, as compared with 74% for all parasitological tests together. The T. vivax and T. congolense primers yielded no positive PCR results.

Glial fibrillary acidic protein and cytokeratin in choroid plexus tumors. An immunohistochemical study.

Ten cases of choroid plexus tumors (3 papillomas and 7 carcinomas) were tested for the presence of glial fibrillary acidic protein (GFAP) and cytokeratin. None of the papillomas and one of the carcinomas were positive with GFAP antisera. Cytokeratin-positive cells were present in 2 of 7 carcinomas and in all papillomas. There seems to be a positive correlation between the degree of the tumor differentiation and the expression of intermediate filaments.

PCR primer evaluation for the detection of trypanosome DNA in naturally infected goats.

The buffy coat of 76 roaming goats from the Bansang and Missira regions in Gambia, was examined for the presence of trypanosomes. From these animals, extractions from dry blood samples on filter paper were subjected to PCR using three different primer sets, ORPHON5J, GOL and TVW, specific for Trypanosoma brucei/Trypanosoma evansi, Trypanosoma congolense and Trypanosoma vivax, respectively. PCR results for T. congolense were 100% concordant with buffy coat examination. Besides the three T. vivax buffy coat-positive samples, another 15 yielded positive with the TVW primers. The ORPHON5J primers yielded no positive results. Analyses with the GOL primers of putatively negative samples, yielded aberrant band patterns whose diagnostic significance still remains to be determined.

Prevalence and incidence of trypanosomosis in horses and donkeys in the Gambia.

In a study of the prevalence and incidence of trypanosomosis in horses and donkeys in two regions of the Gambia, surveys were carried out at Niamina east and Bansang south with a high and low to moderate tsetse challenge, respectively. Eleven horses and 67 donkeys were sampled monthly from August 1997 to September 1998. Blood samples were examined for trypanosomes using the buffy-coat (BC) method and polymerase chain reaction (PCR). Three primer sets were used, specific for either Trypanosoma vivax (TVW), Trypanosoma congolense (GOL) or Trypanosoma brucei (ORPHON5J). The BC results showed that the prevalence (August 1997) and the average monthly incidence (September 1997-1998) of trypanosome infections in horses (45.5 and 16%, respectively) were significantly higher than in donkeys (6.2 and 9%, respectively). Using PCR, the number of detected cases was seven times higher than using the BC. T. congolense was the most frequently observed species, followed by T. vivax and T. brucei. This study confirms earlier observations by other authors that donkeys, which are exposed to a similar tsetse challenge as horses, are significantly less infected with trypanosomes than the latter.

[Evaluation and control of the microbiological quality of hands in foodhandlers]. / Avaliacao e controle da qualidade microbiologica de maos de manipuladores de alimentos.

Microbiological analyses of workers' hands were made for the common indicators, including aerobic mesophilic plate counts (APC), as well as the common food pathogens. Opportunities were observed for cross-contamination of roast beef by workers' hands during slicing operations. Workers' hands showed APC counts of up to 10(7) CFU/hand and the presence of S. aureus and C. perfringens. Salmonella spp were not isolated from hands. These results show that handling of these foods by such workers would be a risk in transmitting pathogenic microorganisms to the foods and is apparent that it is necessary for these workers to take care of personal hygiene. Decimal reductions obtained in the microbiological counts after washing and antisepsis of workers' hands were at 2,6 logs cycles and still demonstrated the importance of this practice in food services by the fact that pathogens such as S. aureus and C. perfringens were inhibited or killed.

[Maternal mortality due to heart disease]. / Mortalidade materna por cardiopatia.

A retrospective study on maternal mortality in pregnant women with cardiac disease over a period of eleven years (January 1979 to December 1989) was undertaken. The objective was an analysis of the main aspects of this association. Cardiac disease was diagnosed in 694 patients (4.2%) of a total of 16,423 admitted to the Obstetrics Department of the Escola Paulista de Medicina. As for etiology, rheumatic disease (52.3%); Chagas's disease (19.3%) and congenital disease (8.1%) were the most frequent causes. There were 51 maternal deaths, according to FIGO's definition (1967), corresponding to a maternal mortality rate of 428.2/100,000 livebirths during the same period. Twelve of these maternal deaths were due to cardiac disease (maternal mortality rate of 100.8/100,000 livebirths). The statistical analysis identified the following aspects associated with maternal mortality among patients with cardiac disease: primigravida, lack of adequate prenatal care, and cardiac surgery performed previously to and/or during pregnancy. Congestive heart failure with pulmonary edema (41.7%) and thromboembolism (25.0%) were the most frequent causes of maternal death among patients with cardiac disease. The NYHA functional classification was not a good parameter for pregnancy prognosis: eleven patients (91.7%) were considered as belonging to the favorable group before they became pregnant. Most maternal deaths occurred during the first 72 hours after delivery. Therefore, this period was considered most critical for maternal mortality in patients with cardiac disease. No relation-ship was found among the factors: maternal age, race, marital status, delivery and maternal mortality among patients with cardiac disease.(ABSTRACT TRUNCATED AT 250 WORDS)

[Maternal mortality in Brazil, 1980]. / Mortalidade materna no Brasil, 1980.

PIP: The authors studied Brazilian maternal mortality by examining official statistics from the country's various geographical regions, including the state of Sao Paulo, Brazil. 1980 was selected because of the possibility of working with data from the Population Census of that year. The principal causes of death in Brazil were hypertension, hemorrhage, puerperal infection, and abortion. In Sao Paulo, where the 4th digit of the IDC is used, the main cause of death was eclampsia. The 2nd was hemorrhage, and the 3rd was infection, whether due to or associated with pregnancy. As for age, the lowest maternal mortality rate occurred in the 20-29 year old age group, the rate was slightly higher for those ages 15-19, and increasing gradually in those ages 30-39 and 40-49. It was not possible to analyze the rates for those ages 10-14 and over age 50 because of lack of data on livebirths. However, there were 18 deaths and 4 deaths respectively in those groups which shows that at opposite poles of the reproductive span, there exists a considerably high mortality rate. In comparing these data with those of other countries, it was found that the majority of deaths are avoidable and that it should be possible to reduce the number considerably by means of better health care for women and by using techniques and resources which are already available. (author's modified)^ieng

[Multicenter study on the prevalence of hepatitis C virus infection in blood donors in the city of Curitiba, Brazil]. / Estudo multicêntrico da prevalência de infecção pelo vírus da hepatite C em doadores de sangue na cidade de Curitiba, Brasil.

In order to check the presence of anti-HCV in the blood of voluntary blood donors in four of the most active blood banks of Curitiba, PR, Brazil a third generation immunoassay (Murex--version III, ABBOTT AxSYM HCV version 3.0) was used. The study detected 347 positive cases, i.e. a median prevalence of 0.80% (range 0.64%-1.11%). Only 44 from the 347 anti-HCV positive blood donors had elevated values of serum aminotransferases (12.68%). Such low values of ALT suggest that the confirmatory RIBA should be applied to the remnant 303 anti-HCV positive cases (87.32%) which did not show elevation of ALT, with the expectation that 30% to 40% of them might be positive. The prevalence of anti-HCV in blood donors in Curitiba seems similar to published data in the Brazilian medical literature.

[Heart surgery with extracorporeal circulation in pregnant women. Analysis of materno-fetal outcome]. / Cirurgia cardíaca com circulação extracorpórea em gestantes. Análise da evolução materno-fetal.

PURPOSE: To analyze maternal and fetal outcome in pregnant undergone to cardiac surgery. METHODS: We studied the evolution of 30 pregnant women submitted to cardiac surgery at the Hospital São Paulo, between Jan/81 and Dec/92 and, further, attended at this Hospital till the parturition. The following variables were analyzed: cardiopulmonary bypass, time of the procedure and time of the anoxia, patient temperature, surgical complications, and neonatal, maternal and fetal outcomes. RESULTS: All patients had rheumatic heart disease and, in 17, mitral stenosis was the main anatomic abnormality. Mitral commissurotomy was performed in 24 patients, double comissurotomy (mitral and aortic) in 1 patient and valve replacement was performed in 5. Cardiopulmonary bypass was utilized in all procedure; occurrence of surgical complications (p < 0.001) and the prolonged surgical time (p = 0.009) were related to the fetal mortality. There was 4 (13.3%) maternal deaths and 10 (33.3%) fetal deaths related to the surgery. CONCLUSION: The indication of cardiac surgery in pregnant women is heart failure, refractory to conventional therapy; cardiopulmonary bypass is associated with high fetal mortality.