Atypical Toxoplasma gondii genotype from a sheep and a pig on Fernando de Noronha Island, Brazil, showed different mouse virulence profiles.

Toxoplasma gondii is a zoonotic parasite which can infect almost all warm-blooded animals. Toxoplasma gondii isolates from Brazil have greater genetic diversity with a predominance of virulent and atypical genotypes, compared with the Northern Hemisphere. Considering that previous studies have demonstrated a high seroprevalence of T. gondii antibodies in animals from Fernando de Noronha Island, the aim of this study was to isolate, genetically characterize, and determine mouse virulence of isolates of T. gondii from livestock from this Brazilian island. Two T. gondii isolates were obtained by mouse bioassay from brain from one sheep and one pig. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22- 8, c29-2, PK1, L358, and Apico) and an atypical genotype of T. gondii (ToxoDB #146) was identified for both isolates. Genotyping of four ROP loci indicated different alleles for ROP16 and mouse virulence analysis revealed different profiles (intermediate and low virulence). This is the first report of this genotype being described in a pig and a sheep.

Dermoscopic and reflectance confocal microscopic presentation of Hailey-Hailey disease: A case series.

BACKGROUND/PURPOSE: Hailey-Hailey disease is a rare inherited acantholytic skin disorder characterized by heterogeneous clinical presentation. Its differential diagnosis might be wide, including other genodermatoses, inflammatory, and infectious skin diseases. Although histopathology remains as diagnostic gold standard, noninvasive techniques such as dermoscopy and reflectance confocal microscopy may assist clinical examination. Herein, we aim to further characterize the dermoscopic and reflectance confocal microscopic presentation of Hailey-Hailey disease with histologic correlation. METHODS: Eight patients with Hailey-Hailey disease were consecutively recruited. All patients were examined using dermoscopy and reflectance confocal microscopy. RESULTS: In all cases, dermoscopy enabled the visualization of polymorphous vessels, including glomerular and linear-looped vessels, within a pink-whitish background. Reflectance confocal microscopy revealed wide suprabasilar partial acantholysis and clefting, crusts, dilated papillae with tortuous vessels, and inflammatory cells. Dyskeratosis, uplocated papillae, and adnexal sparing were also observed. CONCLUSION: Although definite diagnosis was obtained by histopathology in all cases, dermoscopy and reflectance confocal microscopy allowed the identification of common features (even in cases with dissimilar clinical presentation) that may support an early diagnosis of Hailey-Hailey disease, and its differentiation from other more frequent skin disorders.

Biofilm formation on different pH conditions by Streptococcus agalactiae isolated from bovine mastitic milk.

Streptococcus agalactiae is among the most relevant aetiologic agent of bovine clinical and subclinical mastitis, a major problem for the dairy industry. In Brazil, clonal diversity, capsular typing and multidrug resistance profiles of S. agalactiae related to human and bovine infections need further investigation. Presently, S. agalactiae isolates of bovine subclinical mastitis, from Brazilian Northeastern region, were submitted to capsular and pulsed-field gel electrophoresis (PFGE)-typing, antimicrobial susceptibility and assays of biofilm formation at different time incubation and pH levels. Sixteen bovine isolates were characterized by polymerase chain reaction assay as S. agalactiae capsular type II (CTII) and classified by PFGE in A1/A2 (n = 06), B1/B2 (n = 06), C (n = 03) and D (n = 01) patterns. Bovine S. agalactiae CTII strains were classified as 25% multidrug-resistant (MDR) with susceptibility to penicillin, linezolid and vancomycin. Biofilm formation on abiotic surface was strain- and time-dependent with significantly higher rates at pH 6·5. In conclusion, S. agalactiae capsular type II isolates recovered from bovine subclinical mastitis produced different pH-dependent biofilm levels. Our findings suggest that biofilm production is modulated by environmental factors and provides S. agalactiae advantageous in colonizing mammary gland during mastitis development, including MDR strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Streptococcus agalactiae is among the most relevant aetiologic agent of bovine clinical and subclinical mastitis, a major problem for the dairy industry. The disease may cause significant economic loss due to decreased production and milk quality and increased use of medicaments. Presently, data demonstrated that biofilm formation favours the establishment of infectious process in health mammary tissue by S. agalactiae and emphasizes that an acidic pH promotes adhesion by biofilm-forming bacterial strains. S. agalactiae strains (25%) showed resistance to tetracycline, azithromycin, erythromycin and clindamycin, and consequently were classified as multidrug-resistant strains.

Streptococcus agalactiae infection in cancer patients: a five-year study.

Although the highest burden of Streptococcus agalactiae infections has been reported in industrialized countries, studies on the characterization and epidemiology are still limited in developing countries and implementation of control strategies remains undefined. The aim of this retrospective study was to assess the epidemiological, clinical, and microbiological aspects of S. agalactiae infections in cancer patients treated at a Reference Brazilian National Cancer Institute - INCA, Rio de Janeiro, Brazil. We reviewed the clinical and laboratory records of all cancer patients identified as having invasive S. agalactiae disease during 2010-2014. The isolates were identified by biochemical analysis and tested for antimicrobial susceptibility. A total of 263 strains of S. agalactiae were isolated from cancer patients who had been clinically and microbiologically classified as infected. S. agalactiae infections were mostly detected among adults with solid tumors (94 %) and/or patients who have used indwelling medical devices (77.2 %) or submitted to surgical procedures (71.5 %). Mortality rates (in-hospital mortality during 30 days after the identification of S. agalactiae) related to invasive S. agalactiae infections (n = 28; 31.1 %) for the specific category of neoplasic diseases were: gastrointestinal (46 %), head and neck (25 %), lung (11 %), hematologic (11 %), gynecologic (4 %), and genitourinary (3 %). We also found an increase in S. agalactiae resistance to erythromycin and clindamycin and the emergence of penicillin-less susceptible isolates. A remarkable number of cases of invasive infections due to S. agalactiae strains was identified, mostly in adult patients. Our findings reinforce the need for S. agalactiae control measures in Brazil, including cancer patients.

A study on the protein fraction of coffee silverskin: Protein/non-protein nitrogen and free and total amino acid profiles.

Silverskin, the main coffee roasting by-product, is rich in fiber, protein and antioxidants. Its protein fraction was studied regarding total, protein and non-protein nitrogen content. Amino acids were analysed after automated on-line derivatization. The method showed to be precise (<6.9%) and accurate (recoveries using a certified reference material and spiked blanks: 90-102%; for spiked samples: 73-113%). The real protein content of silverskin was 12%. One quarter of the total nitrogen corresponded to the non-protein fraction. All essential amino acids were present in the free form, except methionine. Regarding total amino acids, aspartic and glutamic acids (9-10 mg/g) were the major compounds. Branched chain amino acids (leucine, isoleucine and valine) were also present in substantial amounts (5-8 mg/g), as well as proline and arginine (~5 mg/g). Concluding, silverskin is a source of amino acids with relevance for the improvement of cognitive and physical performances.

Multicaloric effect in a multiferroic composite of Gd5(Si,Ge)4 microparticles embedded into a ferroelectric PVDF matrix.

The coupling between electric, magnetic and elastic features in multiferroic materials is an emerging field in materials science, with important applications on alternative solid-state cooling technologies, energy harvesting and sensors/actuators. In this direction, we developed a thorough investigation of a multiferroic composite, comprising magnetocaloric/magnetostrictive Gd[Formula: see text]Si[Formula: see text]Ge[Formula: see text] microparticles blended into a piezo- and pyroelectric poly(vinylidene) fluoride (PVDF) matrix. Using a simple solvent casting technique, the formation and stabilization of PVDF electroactive phases are improved when the filler content increases from 2 to 12 weight fraction (wt.%). This effect greatly contributes to the magnetoelectric (ME) coupling, with the ME coefficient [Formula: see text] increasing from 0.3 V/cm.Oe to 2.2 V/cm.Oe, by increasing the amount of magnetic material. In addition, magnetic measurements revealed that the ME-coupling has influenced the magnetocaloric effect via a contribution from the electroactive polymer and hence leading to a multicaloric effect. These results contribute to the development of multifunctional systems for novel technologies.

Comparative virulence of Caribbean, Brazilian and European isolates of Toxoplasma gondii.

BACKGROUND: Toxoplasma gondii is a zoonotic parasite of global importance. The outcome of infection in humans can depend on a number of factors including the infecting stage of the parasite, inoculating dose and virulence of the infecting strain. Molecular epidemiological studies have demonstrated an abundance of atypical strains of T. gondii in South America, many of which have been associated with more severe sequelae of infection. The aim of this study was to compare the virulence of T. gondii strains isolated in the Caribbean to a virulent Brazilian strain and an avirulent European strain. METHODS: One hundred and twenty Swiss CD-1 mice were split into 8 groups of 15 mice and each group was inoculated with 200 tachyzoites of one of 8 isolates, comprising ToxoDB genotypes #1, #141, #265, #13, #3 and #6. Five mice per group were euthanized at day 8 post-inoculation (p.i.) and parasite burden was determined in heart, lungs and eyes using quantitative PCR. Lungs and brain were also examined by histopathology and immunohistochemistry. The remaining 10 mice per group were part of a survival experiment to assess virulence. DNA was extracted from tachyzoites of each of the 8 T. gondii isolates and genotyped at four ROP gene loci, including ROP5, ROP16, ROP17 and ROP18 to look for association with markers of virulence. RESULTS: Infection with ToxoDB genotype #13 from the Caribbean resulted in 100% of mice being euthanized which was comparative to infection with the virulent Brazilian strain (ToxoDB genotype #6). Significantly higher parasite burdens were recorded in the lungs and eyes of mice infected with ToxoDB genotypes #13 and #6. Genotyping of ROP loci revealed that the virulent Caribbean isolates had a different ROP18/ROP5 allelic profile (3/1) to the virulent Brazilian isolate (1/3); however, the avirulent Caribbean isolate (ToxoDB genotype #1) had the same ROP18/ROP5 profile as the avirulent European isolate (ToxoDB #3) (both 2/2). Caribbean isolates of intermediate virulence (ToxoDB #141 and #265) all had the same ROP18/ROP5 allelic profile (2/2). CONCLUSIONS: Isolates from the Caribbean with ToxoDB genotype #13 were acutely virulent for mice and comparable to a known virulent Brazilian isolate. The ROP protein allelic profile of the virulent Caribbean and Brazilian isolates differed indicating that perhaps other factors are involved in predicting virulence. Understanding virulence is important for predicting disease outcome in humans and may also aid vaccine design as well as drug discovery.

c-Raf regulates cell survival and retinal ganglion cell morphogenesis during neurogenesis.

The signaling cascade Ras/Raf/mitogen-activated protein kinases modulates cell proliferation, differentiation, and survival, all key cellular processes during neural development. To better define the in vivo role of Raf during chick retinal neurogenesis, we interfered with Raf-dependent signaling during days 4.5 to 7.5 of embryonic development by expressing a dominant negative mutant of c-Raf (DeltaRaf), which blocks Ras-dependent Raf activation, and by overexpressing wild-type c-Raf. DeltaRaf expression induced an increase in cell death by apoptosis, whereas it did not affect overall cell proliferation and differentiation. In parallel, the number of Islet-1/2-positive and TUJ1-positive retinal ganglion cells were diminished in their definitive layer, whereas there was an increase in the number of mislocated Islet-1/2-positive cells. This disturbed morphogenesis correlated with a disruption of the optic fiber layer. Conversely, c-Raf overexpression caused moderate opposite effects on apoptosis. These results frame in vivo early neurogenesis processes in which c-Raf is essential.

Occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in raw milk of sheep and goats of local breeds reared in Northeastern Brazil.

The aim of the present study was to detect Toxoplasma gondii DNA in raw milk samples of goats and sheep of local breeds from the semi-arid region of the states of Pernambuco and Paraíba, Northeastern Brazil. Serum and milk samples were collected from 243 animals (186 goats and 57 sheep). The Indirect fluorescent antibody test (IFAT) was used to search for anti-T. gondii IgG antibodies with a cutoff of 64. Subsequently, the raw milk samples were subjected to DNA extraction and PCR to detect DNA of T. gondii. The IFAT results showed a 6.58% (16/243) positivity when all the samples were considered and a positivity of 15.78% (9/57) and 3.76% (7/186) for goats and sheep samples, respectively. The PCR assay detected T. gondii DNA in 2.06% (5/243) of all the samples tested. All the PCR positive samples were from goats. This result shows the importance of adopting measures of flock's sanitary management and avoiding the consumption of raw milk may constitute a potential risk to the health of milk consumers in this region.

Dietary nucleotide effects upon immune function in infants.

Nucleotide (NT) nitrogen, a component of nonprotein nitrogen, accounts for approximately 0.1% to 0.15% of the total nitrogen content of human milk. The results of studies in animals indicate that dietary NTs may be required for maintenance of normal immune function. Thirty-seven healthy term infants were either breast-fed (n = 9) or fed SMA formula supplemented with 33 mg of NTs per liter (n = 13, NT+) or standard SMA formula (n = 15; NT-). At 2 months of age, natural killer cell percent cytotoxicity was significantly higher in the breast-fed and NT+ groups compared with the NT- group (41.7 +/- 4.7, 32.2 +/- 3.4, 21.7 +/- 2.2%, respectively). Interleukin-2 production by stimulated mononuclear cells was higher in the NT+ compared with the NT- group at 2 months of age (0.90 +/- 0.28 U/mL, 0.27 +/- 0.11 U/mL, respectively); neither formula-fed group differed significantly from the breast-fed group. Rate of growth and incidence and severity of infections did not differ significantly among dietary groups. Nucleotides may be a component of human milk that contributes to the enhanced immunity of the breast-fed infant.

Infant mortality statistics do not adequately reflect the impact of short gestation.

OBJECTIVE: To determine the accuracy of National Center for Health Statistics cause-specific infant mortality data. The National Center for Health Statistics compiles these data by applying World Health Organization (WHO) selection rules to death certificate data. The WHO rules arrange medical entities into a hierarchical order and select a single underlying cause of death (ULCD). DESIGN: A comparison was made between ULCD assigned to a cohort of 335 infant death certificates by (1) a neonatologist panel, and (2) application of WHO selection rules by the Florida State Office of Vital Statistics (OVS). RESULTS: Among the 146 infants with birth weights 1500 to 4649 g, agreement between neonatologists and OVS was 88%, while among the 189 infants with birth weights 0 to 1499 g (very low birth weight), agreement was just 41%. Neonatologists selected short gestation as the underlying cause of death for 82% of very low birth weight infant records, vs 25% by OVS. Due to the application of specific WHO selection rules, OVS frequently selected immediate causes of death, such as "cardiac arrest," as the ULCD, even when the medical certifier had indicated short gestation as the ULCD. In vital statistics reports, many of these immediate causes are reported as "other respiratory conditions of newborn," or "all other perinatal conditions." CONCLUSION: WHO ULCD selection rules should be modified to allow short gestation to have a higher priority over immediate causes of infant death.

(Pro)insulin and insulin-like growth factor I complementary expression and roles in early development.

Evidence that the insulin-like growth factors play a role in embryonic as well as postnatal growth and central nervous system development has accumulated recently from studies using knock-out mice models. However, no effects of IGF-I and II have been demonstrated prior to organogenesis in these studies. We summarize here results supporting the role of insulin (or its precursor proinsulin) in vertebrate development prior to the expression of IGFs. (Pro)insulin mRNA is expressed in the chick embryo during neurulation and early organogenesis and its inhibition by antisense oligodeoxynucleotides increase apoptosis. In another system, proliferative neuroretina, (pro)insulin expression predominates over IGF-I expression. Modulation of apoptosis by (pro)insulin in retina may be largely responsible for the observed stimulation of DNA synthesis and neuronal differentiation. These effects are elicited as well by IGF-I, expressed later in neuroretina. Thus, these polypeptides have complementary expression in early embryos which suggests coordinated actions during development.

Ultrasound evaluation of patients with carpal tunnel syndrome before and after endoscopic release of the transverse carpal ligament.

AIM: To evaluate the cross-sectional area of the median nerve using ultrasound in carpal tunnel syndrome patients before and after endoscopic intervention. MATERIAL AND METHODS: Twenty patients with carpal tunnel syndrome (15 women and five men; mean age 55 years) were prospectively evaluated. Informed consent was obtained from all participants. The study was approved by our Institutional Review Board (IRB). Median nerve cross-sectional area was evaluated at the proximal level before and at 4, 8, and 12 weeks after endoscopic release of the transverse ligament. In the present study, the median nerve cross-sectional area cut-off point was 10 mm(2). Repeated measures analysis of variance test (ANOVA) was applied to compare the reproducibility of ultrasound measurements before and after intervention. RESULTS: The mean cross-sectional area of the median nerve was 15 mm(2) (SD+/-2.1) before surgery; and 11.1 mm(2) (SD+/-3); 9.2 mm(2) (SD+/-2); and 8.6 mm(2) (SD+/-1.6) at 4, 8, and 12 weeks after surgery. Repeated measures analyses of variance were found to be statistically significant (p<0.001). CONCLUSION: The results of the present study demonstrated that there was a decrease in the cross-sectional area of the median nerve after the release of the transverse carpal ligament.

Relativistic quantum thermodynamics of ideal gases in two dimensions.

In this work we study the behavior of relativistic ideal Bose and Fermi gases in two space dimensions. Making use of polylogarithm functions we derive a closed and unified expression for their densities. It is shown that both type of gases are essentially inequivalent, and only in the non-relativistic limit the spinless and equal mass Bose and Fermi gases are equivalent as known in the literature.