Molecular cloning and characterization of a novel ABC transporter gene in the human pathogen Trichophyton rubrum.

A gene encoding an ABC transporter in the dermatophyte Trichophyton rubrum, TruMDR1, was cloned by PCR using degenerate primers. The open reading frame of TruMDR1 is 4838 bp long and the deduced amino acid sequence shows high homology with ABC transporters involved in drug efflux of other fungi. The effect of chemicals on the expression level of mRNAs of this gene was analysed by Northern blot. An increase in expression level was observed when the fungus was exposed to ethidium bromide, ketoconazole, cycloheximide, fluconazole, griseofulvin, imazalil and itraconazole, suggesting the participation of this gene in drug efflux in this dermatophyte. The identification of a gene potentially involved in cellular detoxification in a pathogenic fungus is the first step towards knowing molecular events related to antifungal resistance.     

Diabetic Foot Disease: Emerging Technologies: Repeatability of Infrared Plantar Thermography in Diabetes Patients: A Pilot Study

Objective

Infrared (IR) thermography has been used as a complementary diagnostic method in several pathologies, including distal diabetic neuropathy, by tests that induce thermoregulatory responses, but nothing is known about the repeatability of these tests. This study aimed to assess the repeatability of the rewarming index in subjects with type 2 diabetes mellitus (T2DM) and nondiabetic control subjects.

Methods

Using an IR camera, plantar IR images were collected at baseline (pre-) and 10 min after (post-) cold stress testing on two different days with 7 days interval. Plantar absolute average temperatures pre- and post-cold stress testing, the difference between them (ΔT), and the rewarming index were obtained and compared between days. Repeatability of the rewarming index after the cold stress test was assessed by Bland–Altman plot limits of agreement.

Results

Ten T2DM subjects and ten nondiabetic subjects had both feet analyzed. Mean age did not differ between groups (p = .080). Absolute average temperatures of plantar region pre- (p = .033) and post-cold stress test (p = .019) differed between days in nondiabetic subjects, whereas they did not differ in T2DM subjects (pretest, p = .329; post-test, p = .540). ΔT and rewarming index did not differ between days for both groups, and the rewarming index presented a 100% agreement of day-to-day measurements from T2DM subjects and 95% with nondiabetic subjects.

Conclusions

The rewarming index after cold stress testing presented good repeatability between two days a week in both groups. Despite T2DM subjects presenting no differences on absolute temperature values between days, ΔT or rewarming index after cold stress testing remain recommended beside absolute temperature values for clinical use.     

Apical transportation: Two assessment methods

Root canal transportation can lead to treatment failure. A large number of methodologies for assessing root canal preparation have been tried in the past. This study compared two methods for apical transportation measurement: digitised images of longitudinal root sections and radiographs. Sixty upper molar mesiobuccal root canals prepared for endodontic treatment were assessed. The results did not demonstrate statistically significant differences between the two imaging methods used to evaluate root canal transportation. The two methods were proven to be equally reliable.     

The experience of patients undergoing awake craniotomy: in the patients' own words. A qualitative study

Intraoperative mapping is a well-established and safe technique to maximize the excision of tumors involving the eloquent cortex while minimizing neurological damage. Although different techniques are used, very little has been documented about how the patients feel, what they think about, or how they approach this type of surgery without an anesthetic. Considering the inconsistency of literature, a phenomenology study was conducted with the purpose to describe the human experience of patients before, during, and immediately after awake craniotomy. Twenty-one patients were interviewed at length, both the evening before and the evening after surgery. Awake craniotomy is a complex and subjective experience, the behavioral patterns of the subjects interviewed can be attributed to the instinct of self-preservation, the ability to participate during the procedure helping the surgeon in avoiding brain damage; to be in control of the situation; and to reassure themselves and others. It is most important for a healthcare team to understand the patients' experiences. Immediately before and after surgery, the patients seem to concentrate more on keeping their emotions in check and focus on the risk of subsequent defect or disability, rather than on the brain cancer and what it may imply on life. During the surgery, they become particularly involved in the task: they feel directly responsible for the results of surgery relating this to the effectiveness of collaboration with the neurosurgeons.     

Using cDNA microarrays to identify human CD19(+) B cell gene products (ESTs) originated from systemic lupus erythematosus susceptibility loci

Systemic lupus erythematosus (SLE) is a prototype of autoimmune disease which arises from interactions between susceptibility genes and environmental factors. Despite the heterogeneous manifestations in this disease, all SLE patients present plasma autoantibodies recognizing nuclear components. Thus, auto reactive B cells represent key effectors to be investigated. Human linkage analysis is providing the localization of susceptibility loci distributed in chromosomes contributing to elucidate the manner in which interactions between these loci mediate SLE pathogenesis. We associate the cDNA microarray technology to investigate the differential gene expression of CD19(+) B cells with genetic linkage data. Bioinformatics programs served to evidentiate the differentially expressed sequences and the design of the microarray allowed hierarchical clustering of patients and controls. Sequencing allowed the identification of 8 new gene products differentially expressed (ESTs) that were co-localized in SLE or other autoimmune diseases susceptibility loci on chromosome 1p21, 2q21, 13q33, 16p12.1 and 16q12.1. These findings strongly suggest that chromosomal regions previously identified as SLE susceptibility loci are in fact transcribed in CD19(+) B cells of patients. In this review, we delineate a new possibility for the use of cDNA microarrays in studies focusing the control of gene expression of disease susceptibility loci identified by genetic linkage.     

Anterior sagittal approach without splitting the rectal wall

INTRODUCTIONThe anterior sagittal transrectal approach (ASTRA) has already become popular to treat lesions in the proximal urethra such as trauma, duplicity and stenosis, prostatic utricle, urethral–vaginal fistulas and urogenital sinus anomalies. It provides much better exposure than the traditional perineal approach. Morbidity caused by this technique could be potentially decreased if the anterior sagittal access were to be made without sectioning the rectum. We report our initial experience using anterior approach without rectal sectioning for the treatment of three different types of pelvic disorders.PRESENTATION OF CASEAnterior sagittal access without sectioning the rectal wall was carried out in three different clinical cases – a vaginoplasty in a female patient with congenital adrenal hyperplasia; to treat paradoxical urinary incontinence in a patient with proximal hypospadias (46XY karyotype) and another one with gonadal dysgenesis (46XO/XY karyotype).DISCUSSIONSeveral surgical techniques have been reported to repair congenital or acquired lesions in the posterior urethra with high morbidity and no guarantees of adequate and safe surgical exposition. ASTRA provides an excellent exposure, splitting only the anterior rectal wall. In this study, the anterior sagittal approach was applied without splitting the rectal wall to repair different posterior urethral anomalies, providing excellent exposure without compromising the fecal continence mechanism.CONCLUSIONThe anterior sagittal approach without splitting the rectum is a feasible procedure which provides excellent exposure to the posterior urethra in most cases and leads to less morbidity as it avoids the splitting and suturing of the rectum anterior wall.     

Influence of catechol-O-methyltransferase (COMT) gene polymorphisms in pain sensibility of Brazilian fibromialgia patients

Fibromyalgia syndrome (FS) is a rheumatic syndrome affecting to 2–3% of individuals of productive age, mainly women. Neuroendocrine and genetic factors may play a significant role in development of the disease which is characterized by diffuse chronic pain and presence of tender points. Several studies have suggested an association between FS, especially pain sensitivity, and polymorphism of the catechol-O-methyltransferase (COMT) gene. The aim of the present study was to characterize the SNPs rs4680 and rs4818 of the COMT gene and assess its influence in pain sensitivity of patients with fibromyalgia screened by the Fibromyalgia Impact Questionnaire (FIQ). DNA was extracted from peripheral blood of 112 patients with fibromyalgia and 110 healthy individuals and was used as template in PCR for amplification of a 185-bp fragment of the COMT gene. The amplified fragment was sequenced for analyses of the SNPs rs4680 and rs4818. The frequency of mutant genotype AA of SNP rs6860 was 77.67% in patients with FS and 28.18% for the control group. For the SNP rs4818, the frequency of mutant genotype CC was 73.21 and 39.09% for patients with FS and controls, respectively. Moreover, the FIQ score was higher in patients with the homozygous mutant genotype for SNPs rs4680 (87.92 points) and rs4818 (86.14 points). These results suggest that SNPs rs4680 and rs4818 of the COMT gene may be associated with fibromyalgia and pain sensitivity in FS Brazilian patients.     

Validation of a Brazilian quantitative sensory testing (QST) device for the diagnosis of small fiber neuropathies

Quantitative sensory testing (QST) is defined as the determination of thresholds for sensory perception under controlled stimulus. Our aim was to validate a new QST device for Brazilian sample. In 20 healthy adults, thermoalgesic thresholds were assessed using a QST prototype (Heat Pain Stimulator-1.1.10; Brazil). A 30 × 30 mm(2) thermode with a 1°C/s stimulus change rate were applied. Thresholds of three consecutive stimuli were averaged in two different sessions separated by at least two weeks. Additionally long thermal heat pain stimulus was performed. To evaluate the consistency of our method we also analyzed 11 patients with small fiber neuropathy. Results showed good reproducibility of thermal perception thresholds in normal individuals and plausible abnormal thresholds in patients. We conclude that our QST device is reliable when analyzing the nociceptive pathway in controls and patients.     

Is it correct to always consider weight-bearing asymmetrically distributed in individuals with hemiparesis?

Injuries may cause unilateral deterioration of brain areas related to postural control resulting in lateralized motor disability with abnormal asymmetry in weight-bearing distribution. Although overloading toward the nonaffected limb has been described as the preferred posture among individuals with hemiparesis, characterization of the weight-bearing asymmetry is poorly and indirectly described. Therefore, this study aimed to describe weight-bearing distribution during upright stance, establishing criteria to consider asymmetry in hemiparesis when analyzed within the limits defined by controls matched by age and gender. Forty subjects with (n?=?20) or without hemiparesis (n?=?20) were included in procedures to record weight-bearing values between hemibodies, and these values were used to calculate a symmetry ratio. Control presented 95% confidence interval (CI) of the mean for symmetry ratio ranging from 0.888 to 1.072, defining limits to symmetry. Four subjects with hemiparesis (20%) had symmetry ratios inside limits defined by controls (i.e., weight-bearing symmetrically distributed), and 11 (55%) subjects without hemiparesis showed symmetry ratios outside the limits, suggesting asymmetrical weight-bearing distribution. It was concluded that asymmetry, when present in a control group, was more frequently overloading nonpredominantly used hemibody (nondominant side), differing from a hemiparesis group commonly forced to assume the nonaffected side as the predominantly used hemibody and where the overload was observed.