Characterization of CCDC103 expression profiles: further insights in primary ciliary dyskinesia and in human reproduction.

PROPOSE: To study CCDC103 expression profiles and understand how pathogenic variants in CCDC103 affect its expression profile at mRNA and protein level. METHODS: To increase the knowledge about the CCDC103, we attempted genotype-phenotype correlations in two patients carrying novel homozygous (missense and frameshift) CCDC103 variants. Whole-exome sequencing, quantitative PCR, Western blot, electron microscopy, immunohistochemistry, immunocytochemistry, and immunogold labelling were performed to characterize CCDC103 expression profiles in reproductive and somatic cells. RESULTS: Our data demonstrate that pathogenic variants in CCDC103 gene negatively affect gene and protein expression in both patients who presented absence of DA on their axonemes. Further, we firstly report that CCDC103 is expressed at different levels in reproductive tissues and somatic cells and described that CCDC103 protein forms oligomers with tissue-specific sizes, which suggests that CCDC103 possibly undergoes post-translational modifications. Moreover, we reported that CCDC103 was restricted to the midpiece of sperm and is present at the cytoplasm of the other cells. CONCLUSIONS: Overall, our data support the CCDC103 involvement in PCD and suggest that CCDC103 may have different assemblies and roles in cilia and sperm flagella biology that are still unexplored.

Expression of Estrogen Receptors Alpha (ER-α), Beta (ER-ß), and G Protein-Coupled Receptor 30 (GPR30) in Testicular Tissue of Men with Klinefelter Syndrome.

Men with Klinefelter syndrome (KS) present severe hormonal dysregulation, particularly elevated serum estradiol concentration. Estrogens act through specific receptors and regulate testes development and spermatogenesis. Herein, we evaluated GPR30, ERα, and ERß mRNA expression in testis of KS men and men with 46XY karyotype by reverse transcriptase and quantitative PCR. ERß transcripts are the most abundant in testicular tissue of 46XY men. Notably, testicular GPR30 transcription in KS men was approximately 12 times higher. Since GPR30 is essential to mediate estrogen effects over steroidogenesis, our data illustrate that GPR30 may underpin the testicular alterations observed in KS men.

Combined effects of lactoferrin and lysozyme on Streptococcus pneumoniae killing.

Streptococcus pneumoniae is a common colonizer of the human nasopharynx, which can occasionally spread to sterile sites, causing diseases such as otitis media, sinusitis, pneumonia, meningitis and bacteremia. Human apolactoferrin (ALF) and lysozyme (LZ) are two important components of the mucosal innate immune system, exhibiting lytic effects against a wide range of microorganisms. Since they are found in similar niches of the host, it has been proposed that ALF and LZ could act synergistically in controlling bacterial spread throughout the mucosa. The combination of ALF and LZ has been shown to enhance killing of different pathogens in vitro, with ALF facilitating the latter action of LZ. The aim of the present work was to investigate the combined effects of ALF and LZ on S pneumoniae. Concomitant addition of ALF and LZ had a synergistic killing effect on one of the pneumococci tested. Furthermore, the combination of ALF and ALZ was more bactericidal than lysozyme alone in all pneumococcal strains. Pneumococcal surface protein A (PspA), an important vaccine candidate, partially protects pneumococci from ALF mediated killing, while antibodies against one PspA enhance killing of the homologous strain by ALF. However, the serological variability of this molecule could limit the effect of anti-PspA antibodies on different pneumococci. Therefore, we investigated the ability of anti-PspA antibodies to increase ALF-mediated killing of strains that express different PspAs, and found that antisera to the N-terminal region of PspA were able to increase pneumococcal lysis by ALF, independently of the sequence similarities between the molecule expressed on the bacterial surface and that used to produce the antibodies. LF binding to the pneumococcal surface was confirmed by flow cytometry, and found to be inhibited in presence of anti-PspA antibodies. On a whole, the results suggest a contribution of ALF and LZ to pneumococcal clearance, and confirm PspA's ability to interact with ALF.

Adult circumcision and male sexual health: a retrospective analysis.

We aimed to evaluate possible associations of circumcision with several sexual dysfunctions and to identify predictors for the development of these outcomes post-operatively. Telephone surveys about sexual habits and dysfunctions before and after intervention were conducted post-operatively to patients that underwent circumcision in Centro Hospitalar Vila Nova de Gaia/Espinho during 2011. McNemar test was used for a matched-pairs analysis of pre- and post-operative data. Odds ratios, adjusted in a multivariate analysis, explored predictors of de novo sexual dysfunctions after circumcision. With intervention, there was an increase in frequency of erectile dysfunction (9.7% versus 25.8%, P = 0.002) and delayed orgasm (11.3% versus 48.4%, P < 0.001), and a significant symptomatic improvement in patients with pain with intercourse (50.0% versus 6.5%, P < 0.001). Significant predictors for de novo erectile dysfunction were diabetes mellitus (OR 9.81, P = 0.048) and lack of sexual desire (OR 8.76, P = 0.028). Less than three sex partners (OR 7.04, P = 0.007) and low sexual desire (OR 7.49, P = 0.029) were significant predictors for de novo delayed orgasm.

Caspase-3 detection in human testicular spermatozoa from azoospermic and non-azoospermic patients.

The apoptotic mechanisms underlying spermatogenesis in testis are poorly understood. In the present study, the rates of testicular spermatozoa with active caspase-3 were quantified in testicular samples with normal and impaired spermatogenesis. Testicular spermatozoa were collected from 18 men after testicular biopsy during assisted reproductive treatments: five presented oligozoospermia, four congenital bilateral absence of the vas deferens (CBAVD), five secondary obstructive azoospermia (sOAZ) and four hypospermatogenesis. Ejaculated samples were derived from six normozoospermic patients. Testicular spermatozoa were analysed using a fluorescence microscope and differences among groups were calculated using regression logistic models. Total rates of spermatozoa with active caspase-3 were significantly higher in sOAZ (78.6±13.9), followed by hypospermatogenesis (70.8±5.8), CBAVD (55.9±25.5), oligozoospermia (31.7±31.0) and normozoospermia (20.4±15.5). Distinct patterns of active caspase-3 were observed in testicular spermatozoa compartments: midpiece, equatorial region, acrosomal vesicle region, nucleus and cytoplasm. Hypospermatogenesis showed active caspase-3 mainly in the midpiece. In CBAVD, sOAZ and oligozoospermia, active caspase-3 was mainly in the nucleus, although no differences were found between oligozoospermia and hypospermatogenesis. In sOAZ, active caspase-3 in the spermatozoa nucleus was 1.89-fold higher than in CBAVD. Results suggest that tubular obstruction may induce nuclear lesions and that disrupted spermatozoa production observed in cases of hypospermatogenesis might be associated with mitochondrial lesions.

Effect of Mouth Rinse Treatments on Bleached Enamel Properties, Surface Morphology, and Tooth Color.

OBJECTIVE:: To evaluate, in vitro, the effect of mouth rinse exposure on bleached enamel. METHODS:: Enamel/dentin bovine blocks (4×4×2 mm) were bleached with 35% hydrogen peroxide (HP) and were submitted to immersion twice daily for 14 days with different rinses (n=10), including those involving: distilled water (C [control]), 225-ppm NaF (FM, Colgate Plax Classic), essential oil (EM, Listerine Tartar Control), 1.5% hydrogen peroxide (HPM, Colgate Plax Whitening), and 2% hydrogen peroxide, pyrophosphates, and 225-ppm NaF (HPM+P, Colgate® Luminous White). The specimens were stored in a remineralizing solution during all experiments. Analyses of color (ΔE, L*, a*, b*) and roughness (Ra) were performed at the baseline, after HP, and after exposure to the rinse. The cross-sectional microhardness (CSMH) and images by scanning electron microscopy (SEM) were assessed at the end. The data were subjected to analysis of variance (ANOVA) (ΔE), repeated measures ANOVA (Ra), and split-plot ANOVA (CSMH), followed by the Tukey test. The L*, a*, and b* values were analyzed by generalized linear models (α=0.05). RESULTS:: Color changes were not statistically different in the groups. Ra increased in all groups after bleaching; however, it was reestablished in C, FM, and HPM+F and increased in EM after 14 days of the rinse. EM and HPM reduced the CSMH values differing from C and promoted alterations on the enamel surface visualized by SEM. CONCLUSION:: The mouth rinses did not affect the whitening efficacy or promote benefits on bleached enamel properties. Moreover, the 1.5% hydrogen peroxide- or essential oil-based mouth rinses affected the bleached enamel properties, promoting an alteration in morphologic surface and mineral loss in depth.

[Pre-natal retroperitoneal mass: teratoma or fetus in fetu?]. / Masa prenatal retroperitoneal: teratoma o fetus in fetu?

BACKGROUND: Several case reports in the literature brought to discussion the distinction criteria between fetus in fetu and teratoma. CASE REPORT: The authors present a case report of a premature newborn boy, evaluated for a capsulated abdominal mass with prenatal diagnosis pointing to possible malignant lesion. A CT scan showed an expansive, retroperitoneal lesion, from kidneys to pelvis, sacrum and coccyx, with solid, cystic and fatty areas; the fine needle aspiration was inconclusive, and he underwent surgery on day 9 with partial resection of the lesion. A new procedure to complete the excision was performed 8 months later. The histopathologic examination of the two fragments showed mature and organized components of nervous system, respiratory and digestive apparatus (liver and pancreas included), thymus, nervous ganglia, skin and adnexa, musculoskeletal and brown adipose tissues; cartilage and several focus of calcification. These components showed a craniocaudal orientation, but no evidence of organoid formation. DISCUSSION: This case presents features of both teratoma and fetus in fetu. The assumption of either one of the diagnosis has prognostic implications, with resulting differences in clinical follow up and expectations about the child's health.

Effect of Toothpaste Use Against Mineral Loss Promoted by Dental Bleaching.

AIM: To investigate the effect of different toothpaste formulations used prior to dental bleaching with 35% hydrogen peroxide (HP) on the mineral content and surface morphology of enamel. METHODS: Seventy bovine enamel blocks (4×4×2 mm) were submitted to in vitro treatment protocols using a toothbrushing machine prior to dental bleaching or a placebo procedure (n=10) as proposed in the following groups: unbleached control (PLA), bleached control (HP), and brushing with differing toothpastes prior to HP bleaching, including: potassium nitrate toothpaste containing sodium fluoride (PN), sodium monofluorophosphate/MFP toothpaste (FT), arginine-carbonate (8% arginine) (PA) or arginine-carbonate (1.5% arginine) toothpaste (SAN), and toothpaste containing bioactive glass (NM). Phosphorus concentration in gel ([P]) was evaluated (µg of P/mg of gel), and the elemental levels (wt%) of Ca, P, and Na as well as the proportion between Ca and P and spectra graphics were determined using an energy-dispersive X-ray spectrometer (EDS). The surface morphology was assessed using scanning electron microscopy (SEM). The data were subjected to analysis of variance and the Tukey test (α=0.05). RESULTS: HP demonstrated the greatest [P] values in gel, being statistically different from PLA. The [P] of NM was statistically similar to PLA. HP showed a significant decrease in the Ca% and Ca/P values when compared to PLA in EDS analysis. PA showed Ca/P values statistically different from HP. In accordance with SEM analysis, the PA, SAN, and NM groups presented a smooth and uniform enamel surface, while HP and FT demonstrated some alterations in morphology. CONCLUSION: The toothpastes containing bioactive glass or arginine carbonate used prior to dental bleaching were effective in protecting enamel against mineral loss promoted by the whitening procedure.

Isolation of bovine immunoglobulins resistant to peptic digestion: new perspectives in the prevention of failure in passive immunization of neonatal calves.

In calves, neonatal mortality and disease susceptibility are greatly influenced by failure in passive immunization, normally provided by colostrum ingestion just after birth. Formulations projected to replace natural colostrum have not been successful, and one of the possible reasons for such failure is that orally administered Ig are probably digested in the gastrointestinal tract, so they are not absorbed as intact functional molecules. With the aim of finding an adequate colostrum substitute, we used columns of immobilized jacalin, a lectin known by its ability to bind O-linked oligosaccharides, to obtain a colostral Ig population putatively protected against enzymatic cleavage by the presence of sugar chains. Immunoglobulin G1 is a major constituent of colostrum Ig bound to jacalin (JB-Ig). This preparation contains 10% of the total colostral Ig and is typically 3 to 6 times more resistant to pepsin digestion than the Ig contained in the fraction that is not bound to jacalin, which presumably does not contain O-glycans. Mass spectrometry analysis demonstrated that the tryptic peptides obtained from JB-Ig and unbound Ig were similar, indicating that their distinct susceptibility to enzyme hydrolysis was associated with differences in their sugar chains. Therefore, the present research suggests that the bovine colostrum JB-Ig has potential application in the immunotherapy of neonatal calves that have not been supplied with colostrum.

Twenty years of liver transplantation in Brazil.

This paper summarizes the 20 years of liver transplantation in Brazil, in the context of the Western world scenario. More than 5000 liver transplantations have been performed in the country since September 1, 1985. The living-donor liver transplantation, one of the landmarks in liver transplantation, was first described by our team in 1989. Brazil is the seventh country in number of liver transplants in the Western world and the first in Latin America. Almost 1000 procedures were performed in 2004, 19% of them involving living donors.

Does inter-tibiofibular graft still have a role in the treatment of lower-limb non-union?

INTRODUCTION: Tibial non-union is a complication that poses a real challenge for surgeons. Several forms of treatment, depending on the type of non-union, have been described. The present study sought to assess results for treatment of tibial non-union by inter-tibiofibular graft (ITFG). MATERIAL AND METHOD: An exhaustive cohort study was performed on the files of 33 patients: 25 male, 8 female; mean age, 44years. Twenty cases involved high-energy trauma. Twenty-four were open fractures. Twenty-two concerned diaphyseal fracture, 10 of which were complex segmental. Eleven concerned distal fracture, including 4 complete articular fractures. There were 17 cases of septic non-union. There were no cases of severe bone defect. ITFG was performed at a mean 8.7 months post-trauma, as first-line treatment in 30 cases and in second line in 3. RESULTS: Thirty-one patients showed bone consolidation, at a mean 7.2 months. The 2 failures resulted from technical error. Trauma kinetics emerged as a risk factor for failure. DISCUSSION: ITFG remains a useful treatment option in tibial non-union, whether infected or not. The present results are comparable with those of the literature. Although the present series comprised only tight non-union, a study of the literature showed that ITFG can treat bone defects up to 4 or 5cm. Functional results showed tibiotalar joint stiffening, due more to immobilization and non-weight-bearing than to syndesmosis. ITFG thus remains relevant to the treatment of tibial non-union. LEVEL OF EVIDENCE: IV, retrospective study.

Electronically addressable nanomechanical switching of i-motif DNA origami assembled on basal plane HOPG.

Here, a pH-induced nanomechanical switching of i-motif structures incorporated into DNA origami bound onto cysteamine-modified basal plane HOPG was electronically addressed, demonstrating for the first time the electrochemical read-out of the nanomechanics of DNA origami. This paves the way for construction of electrode-integrated bioelectronic nanodevices exploiting DNA origami patterns on conductive supports.

New frameshift mutation in the 5alpha-reductase type 2 gene in a Brazilian patient with 5alpha-reductase deficiency.

Male pseudohermaphroditism caused by steroid 5alpha-reductase deficiency is an autosomal recessive disorder. The enzyme steroid 5alpha-reductase 2 (encoded by the SRD5A2 gene) catalyses the conversion of testosterone to dihydrotestosterone, which is required for normal differentiation of the external male genitalia. This report describes the molecular analysis of the 5alpha-reductase type 2 gene in a Brazilian patient who was raised as a female, underwent a reversal of gender role behavior, and is now a married man. This patient is a compound heterozygote bearing an A-->G mutation within exon 2, changing codon 126 from Glu to Arg on one allele and a novel single base deletion (418delT) causing a frameshift mutation at codon 140 in the same exon, on the other allele. This last mutation probably leads to the synthesis of a truncated protein, because a premature termination signal is created at codon 159.

Detection of Gly-196-Ser mutation in 5alpha-reductase type II gene in a Brazilian patient with female assignment and behavior.

We describe the identification of a single base mutation in the 5alpha-reductase type II gene in a Brazilian patient who was reared as female and remained with female behavior and sexual identity.

A prospective and randomized study using ribavirin as monotherapy for the treatment of naïve patients with chronic hepatitis C.

In order to evaluate the response to ribavirin in previously untreated patients with chronic hepatitic C, 39 patients were selected for a double-blind prospective and randomized trial, and divided into two groups: ribavirin-group (19 patients) and placebo-group (20 patients). Ribavirin was administered orally for 24 weeks (600 mg/day, followed by 1,000 mg/day and 1,200 mg/day each one for 8 weeks). After 3 months of drug administration, the patients were evaluated by measuring biochemical, virologic and histologic responses. After this phase, ribavirin was offered to the patients who had received placebo (second phase). The results showed that the patients who received ribavirin showed a higher reduction in serum alanine aminotransferase (ALT) activity than patients in the placebo group. Among the patients in the ribavirin-group, a complete biochemical response (ALT levels normalized) was observed in 3 patients (16%), and a partial response (reduction greater than 50% of the initial value of ALT activity) in 4 (21%). In the 20 patients in the placebo group, only 1 showed a partial response (5%). In the second phase of the study, among 16 patients who received ribavirin, 4 (25%) showed a complete and 5 (31%) a partial biochemical response. HCV-RNA did not become negative in any patient during the two phases. A reduction in the score of portal and lobular activity was observed in patients who received ribavirin, but statistical analysis did not identify differences. This study showed that ribavirin alone induces a biochemical response (ALT reduction) in some patients with chronic hepatitis C, which may be associated with a reduction in hepatic inflammatory activity reduction, but the changes are not sufficient to recommend initial monotherapy with ribavirin.

[Spontaneous intracerebral hematomas: study of 121 cases]. / Hematomas intracerebrais espontâneos: estudo de 121 casos.

Results are reported on the study of 121 patients committed by spontaneous intracerebral haematomas (HIE): mean age 53.4 years, 62.8% males, mean admission time of 36 hours (3 hours to 12 days) from onset; 63.5% were graded over 7 in the Glasgow scale and 81.9% were graded 3 or over in the Boterell scale. The HIE location was 45.5% in the basal ganglia, multilobar in 14.7%, lobar in 22.8%, brain stem in 4%, and cerebellum in 2%. The mean diameter was 46.6 mm (16-93) and mean area was 1422.9 mm2 (60-4818). CSF in 67 cases showed mean opening pressure of 234 mmH20 and mean protein content of 416.9 mg/dl. Treatment was conservative in 107 cases and surgical in 14; 55.8% cases survived. The majority of patients who died were graded 3 in the Boterell scale and below 9 in the Glasgow scale. It was found a statistical correlation between death and: low Glasgow and high Boterell scales grading, motor ocular nerve palsy, motor deficit, decerebration signs, bronchopneumonia, large diameter and area of hematomas. No statistical correlation was found regarding survival and treatment with dexamethasone, antifibrinolytics, anticonvulsants, antihypertensive drugs and diuretics. Use of mannitol, fall in the systolic blood pressure and surgical therapy increased the mortality rate. From 14 patients who underwent surgical drainage, 11 died. The main complication who lead to death was bronchopneumonia. A discussion is made on HIE pathogenesis, localization, present day incidence, clinical findings, death causes, size of hematomas and type of treatment used.

[Early differentiation between Duchenne and Becker muscular dystrophy: clinical, laboratory, electrophysiology, histochemical, and immunohistochemical study of 138 cases]. / Diferenciação precoce entre distrofia muscular de Duchenne e de Becker.

194 clinical, laboratory, electrophysiologic, histological, histochemical and immunohistochemical parameters were studied through statistical analysis in 112 cases of Duchenne muscular dystrophy (DMD) and in 26 cases of Becker muscular dystrophy (BMD). It was found a significant statistical difference (p < 0.05) between the two groups concerning the age of evaluation, beginning of symptoms, difficulty in walking, running, climbing and going downstairs, frequent falling down, support to walk, localized muscle pain, stopping climb stairs, and inability to walk. Muscle biopsy showed statistically significant (p < 0.05) differences between the two groups regarding the intensity of connective tissue and focal adipose tissue proliferation, presence of diffuse rounded atrophic and angulated fibers, diffuse hypertrophic and splitting fibers. There were also differences regarding excessive internal fibers nuclei, hypertrophic types 1 and 2 fibers, angulated atrophic fibers and focal increasing in the NADH-TR, angulated atrophic fibers in non-specific esterase, and accumulated NBT in the periphery of fibers in succinic dehidrogenase. Isolatedly muscle biopsy gave the correct diagnosis in 52.7% of DMD cases and in 69.2% of BMD cases. Dystrophin detection by immunofluorescence (60 cases) showed: absence in 87.0% of fibers in DMD cases, and sarcolemmal membrane discontinuites in all BMD cases. The muscle biopsy diagnosis had an agreement with the dystrophin results in 82.6% of DMD cases and 71.4% of BMD cases.

[Kennedy disease: report of 2 cases]. / Doença de Kennedy: relato de dois casos.

We report two cases of Kennedy's disease (muscle weakness, amyotrophy, intentional tremor, endocrine abnormalities, and denervation signs at electromyography). This entity must be differentiated from other motor neuron disorders by the genetic pattern (X-liked recessive), gynecomastia, testicular atrophy, oligospermia and good prognosis. A discussion about the clinical pattern and evolution is made.

[Linear accelerator radiosurgery]. / Radiocirurgia por acelerador linear.

Radiosurgery is the precise radiation of a known intracranial target with a high dose of energy, sparing the adjacent nervous tissue. Technological advances in the construction of linear accelerators, stereotactic instruments and in computer sciences made this technique easier to perform and affordable. The main indications for radiosurgery are inoperable cerebral vascular malformations, vestibular and other cranial schwannomas, skull base meningiomas, deep seated gliomas and cerebral metastases. More recently, the development of fraccionated stereotactic radiotherapy increased the spectrum of indications to bigger lesions and to those adjacent to critical nervous structures. We present our initial experience in the treatment of 31 patients. An adequate control of the neoplastic lesions was obtained and the adequate time of observation is still needed to evaluate the results in arteriovenous malformations.

Carbon Partitioning in Soybean Infected with Meloidogyne incognita and M. javanica.

Seven-day-old seedlings of two cultivars (Cristalina and UFV ITM1) of Glycine max were inoculated with 0, 3,000, 9,000, or 27,000 eggs of Meloidogyne incognita race 3 or M. javanica and maintained in a greenhouse. Thirty days later, plants were exposed to (1)CO for 4 hours. Twenty hours after (1)CO exposure, the root fresh weight, leaf dry weight, nematode eggs per gram of root, total and specific radioactivity of carbohydrates in roots, and root carbohydrate content were evaluated. Meloidogyne javanica produced more eggs than M. incognita on both varieties. A general increase in root weight and a decrease in leaf weight with increased inoculum levels were observed. Gall tissue appeared to account for most of the root mass increase in seedlings infected with M. javanica. For both nematodes there was an increase of total radioactivity in the root system with increased levels of nematodes, and this was positively related to the number of eggs per gram fresh weight and to the root fresh weight, but negatively related to leaf dry weight. In most cases, specific radioactivities of sucrose and reducing sugars were also increased with increased inoculum levels. Highest specific radioactivities were observed with reducing sugars. Although significant changes were not observed in endogenous levels of carbohydrates, sucrose content was higher than reducing sugars. The data show that nematodes are strong metabolic sinks and significantly change the carbon distribution pattern in infected soybean plants. Carbon partitioning in plants infected with nematodes may vary with the nematode genotype.