Association of asymptomatic spinal cord lesions and atrophy with disability 5 years after a clinically isolated syndrome.

BACKGROUND: Spinal cord pathology is an important substrate for long-term disability in multiple sclerosis (MS). OBJECTIVE: To investigate longitudinal changes in spinal cord lesions and atrophy in patients with a non-spinal clinically isolated syndrome (CIS), and how they relate to the development of disability. METHODS: In all, 131 patients with a non-spinal CIS had brain and spinal cord imaging at the time of CIS and approximately 5 years later (median: 5.2 years, range: 3.0-7.9 years). Brain magnetic resonance imaging (MRI) measures consisted of T2-hyperintense and T1-hypointense lesion loads plus brain atrophy. Spinal cord MRI measures consisted of lesion number and the upper cervical cord cross-sectional area (UCCA). Disability was measured using the Expanded Disability Status Scale (EDSS). Multiple linear regression was used to identify independent predictors of disability after 5 years. RESULTS: During follow-up, 93 (71%) patients were diagnosed with MS. Baseline spinal cord lesion number, change in cord lesion number and change in UCCA were independently associated with EDSS ( R2 = 0.53) at follow-up. Including brain T2 lesion load and brain atrophy only modestly increased the predictive power of the model ( R2 = 0.64). CONCLUSION: Asymptomatic spinal cord lesions and spinal cord atrophy contribute to the development of MS-related disability over the first 5 years after a non-spinal CIS.

The non-genomic effects of endocrine-disrupting chemicals on mammalian sperm.

Exposure to toxicants present in the environment, especially the so-called endocrine-disrupting chemicals (EDCs), has been associated with decreased sperm quality and increased anomalies in male reproductive organs over the past decades. Both human and animal populations are continuously exposed to ubiquitous synthetic and natural-occurring EDCs through diet, dermal contact and/or inhalation, therefore potentially compromising male reproductive health. Although the effects of EDC are likely induced via multiple genomic-based pathways, their non-genomic effects may also be relevant. Furthermore, spermatozoa are transcriptionally inactive cells that can come in direct contact with EDCs in reproductive fluids and secretions and are therefore a good model to address non-genomic effects. This review thus focuses on the non-genomic effects of several important EDCs relevant to mammalian exposure. Notably, EDCs were found to interfere with pre-existing pathways inducing a panoply of deleterious effects to sperm function that included altered intracellular Ca(2) (+) oscillations, induction of oxidative stress, mitochondrial dysfunction, increased DNA damage and decreased sperm motility and viability, among others, potentially jeopardizing male fertility. Although many studies have used non-environmentally relevant concentrations of only one compound for mechanistic studies, it is important to remember that mammals are not exposed to one, but rather to a multitude of environmental EDCs, and synergistic effects may occur. Furthermore, some effects have been detected with single compounds at environmentally relevant concentrations.

High glucose concentrations per se do not adversely affect human sperm function in vitro.

Diabetes mellitus (DM) represents one of the greatest concerns to global health and it is associated with diverse clinical complications, including reproductive dysfunction. Given the multifactorial nature of DM, the mechanisms that underlie reproductive dysfunction remain unclear. Considering that hyperglycemia has been described as a major effector of the disease pathophysiology, we used an in vitro approach to address the isolated effect of high glucose conditions on human sperm function, thus avoiding other in vivo confounding players. We performed a complete and integrated analysis by measuring a variety of important indicators of spermatozoa functionality (such as motility, viability, capacitation status, acrosomal integrity, mitochondrial superoxide production and membrane potential) in human sperm samples after incubation with d- and l-glucose (5, 25, or 50 mM) for 24 and 48 h. No direct effects promoted by 25 or 50 mM d-glucose were found for any of the parameters assessed (P>0.05), except for the acrosome reaction, which was potentiated after 48 h of exposure to 50 mM d-glucose (P<0.05). Interestingly, non-metabolizable l-glucose drastically increased superoxide production (P<0.05) and suppressed sperm motility (P<0.05) and capacitation (P<0.05) after 24 h of treatment, whereas mitochondrial membrane potential (P<0.05), acrosomal integrity (P<0.01) and viability (P<0.05) were later decreased. The overall results suggest that high glucose levels per se do not influence human sperm function in vitro, which stresses the importance of other factors involved in DM pathology. Nevertheless, the absence of metabolizable glucose contributes to a severe impairment of sperm function and thus compromises male fertility.

A longitudinal assessment of sleep variables during exacerbations of chronic obstructive pulmonary disease.

Although sleep disturbance is common in chronic obstructive pulmonary disease (COPD), relatively little is known on the effect of the exacerbation on sleep quality. Accordingly, we longitudinally assessed sleep variables during exacerbations and clinical stability. This is a sub-study of a larger observational analysis. Inclusion criteria were clinically stable COPD and two or more clinical exacerbations in the preceding 12 months. Patients were followed for approximately 6 months and during this time the following were recorded daily: (1) COPD exacerbations, which were determined in two ways, clinically and symptom defined using the exacerbations of chronic pulmonary disease tool (EXACT); (2) daytime sleepiness, which was measured using the Stanford Sleepiness Scale; (3) subjective awakenings, which was measured from a sleep diary; and (4) sleep duration, efficiency, and objective awakenings, which was measured from actigraphy. These variables for exacerbation and non-exacerbation days were compared. Seventeen patients (9 male, age 63 ± 12 years, forced expiratory volume in 1 second 52 ± 20%) entered data over 135 ± 18 days. During this time, 15 patients had 27 symptom-defined exacerbations and 8 had 9 clinically reported exacerbations. Symptom-defined exacerbation days were 26% of the total study days. More daytime sleepiness, decreased total sleep time (TST), and decreased sleep efficiency (SE) were present during exacerbations compared with clinical stability (p < 0.001). These disturbances tended to be greater during clinically reported exacerbations than during unreported events (p < 0.05). Increased daytime sleepiness, less TST, and poorer SE are present during COPD exacerbations.

Similar preferences for ornamentation in opposite- and same-sex choice experiments.

Selection due to social interactions comprises competition over matings (sexual selection stricto sensu) plus other forms of social competition and cooperation. Sexual selection explains sex differences in ornamentation and in various other phenotypes, but does not easily explain cases where those phenotypes are similar in males and females. Understanding such similarities requires knowing how phenotypes influence nonsexual social interactions as well, which can be very important in gregarious animals, but whose role for phenotypic evolution has been overlooked. For example, 'mate choice' experiments often found preferences for ornamentation, but have not assessed whether those are strictly sexual or are general social preferences. Using choice experiments with a gregarious and mutually ornamented finch, the common waxbill (Estrilda astrild), we show that preferences for ornamentation in the opposite-sex also extend to same-sex interactions. Waxbills discriminated between opposite- and same-sex individuals, but most preferences for colour traits were similar when interacting with either sex. Similar preferences in sexual and nonsexual associations may be widespread in nature, either as social adaptations or as by-product of mate preferences. In either case, such preferences may set the stage for the evolution of mutual ornamentation and of various other similarities between the sexes.

Acute calcific periarthritis in a proximal interphalangeal joint of the hand after acute trauma: a rare case.

Acute calcific periarthritis (ACP) in the interphalangeal joints of the hand is rare, with less than 100 cases reported. A rare case of ACP in a proximal interphalangeal (PIP) joint of the hand, in a young black woman, after acute trauma, is presented. She experienced severe pain and limited range of motion, and was medicated with an oral corticoid, which was followed by a rapid resolution of the symptoms. At six months, there were no signs of clinical or radiographic recurrence. Recognition of ACP allows for avoiding unnecessary treatments. In this case, treatment with corticoids might have played a role in a faster recovery.

La periartritis calcificada aguda (PCA) en las articulaciones interfalángicas de la mano es rara, con menos de 100 casos reportados. Se presenta un caso raro de PCA en una articulación interfalángica proximal (IFP) de la mano, en una mujer joven de raza negra, después de un traumatismo agudo. Experimentó dolor intenso y rango de movimiento limitado, y fue medicada con un corticoide oral, lo que fue seguido por una rápida resolución de los síntomas. A los seis meses no hubo signos de recurrencia clínica ni radiológica. El reconocimiento de PCA permite evitar tratamientos innecesarios. En este caso, el tratamiento con corticoides podría haber contribuido a una recuperación más rápida.

Polymorphisms and haplotypes of TOLLIP and MUC5B are associated with susceptibility and survival in patients with fibrotic hypersensitivity pneumonitis.

INTRODUCTION AND OBJECTIVES: Hypersensitivity pneumonitis (HP) is an interstitial lung disease with diverse clinical features that can present a fibrotic phenotype similar to idiopathic pulmonary fibrosis (IPF) in genetically predisposed individuals. While several single nucleotide polymorphisms (SNPs) have been associated with IPF, the genetic factors contributing to fibrotic HP (fHP) remain poorly understood. This study investigated the association of MUC5B and TOLLIP variants with susceptibility, clinical presentation and survival in Portuguese patients with fHP. MATERIAL AND METHODS: A case-control study was undertaken with 97 fHP patients and 112 controls. Six SNPs residing in the MUC5B and TOLLIP genes and their haplotypes were analyzed. Associations with risk, survival, and clinical, radiographic, and pathological features of fHP were probed through comparisons among patients and controls. RESULTS: MUC5B rs35705950 and three neighboring TOLLIP variants (rs3750920, rs111521887, and rs5743894) were associated with increased susceptibility to fHP. Minor allele frequencies were greater among fHP patients than in controls (40.7% vs 12.1%, P<0.0001; 52.6% vs 40.2%, P = 0.011; 22.7% vs 13.4%, P = 0.013; and 23.2% vs 12.9%, P = 0.006, respectively). Haplotypes formed by these variants were also linked to fHP susceptibility. Moreover, carriers of a specific haplotype (G-T-G-C) had a significant decrease in survival (adjusted hazard ratio 6.92, 95% CI 1.73-27.64, P = 0.006). Additional associations were found between TOLLIP rs111521887 and rs5743894 variants and decreased lung function at baseline, and the MUC5B SNP and radiographic features, further highlighting the influence of genetic factors in fHP. CONCLUSION: These findings suggest that TOLLIP and MUC5B variants and haplotypes may serve as valuable tools for risk assessment and prognosis in fibrotic hypersensitivity pneumonitis, potentially contributing to its patient stratification, and offer insights into the genetic factors influencing the clinical course of the condition.

Efficacy and safety of fluoroless ureteroscopy and retrograde intrarenal surgery for the treatment of urolithiasis: A comparative study.

INTRODUCTION: Ureteroscopy (URS) and retrograde intrarenal surgery (RIRS) are traditionally guided by fluoroscopy, but the risks of exposure to ionizing radiation may present a matter of concern for patients and urologists. The aim of this study was to evaluate the efficacy and safety of fluoroless URS and RIRS compared with conventional fluoroscopy-guided procedures for the treatment of ureteral and renal stones. MATERIAL AND METHODS: Patients treated with URS or RIRS for urolithiasis between August 2018 and December 2019 were retrospectively evaluated and grouped according to the use of fluoroscopy. Data was collected from individual patient records. The main outcomes were stone-free rate (SFR) and complications, compared between the fluoroscopy and fluoroless groups. A subgroup analysis by type of procedure (URS and RIRS) and a multivariate analysis to identify predictors of residual stones were conducted. RESULTS: A total of 231 patients met the inclusion criteria: 120 (51.9%) in the conventional fluoroscopy group and 111 (48.1%) in the fluoroless group. No significant differences were found between groups regarding SFR (82.5% vs 90.1%, p=.127) or postoperative complication rate (35.0% vs 31.5%, p=.675). In the subgroup analysis these variables did not present significant differences, regardless of the procedure considered. In the multivariate analysis the fluoroless technique was not an independent predictor of residual lithiasis (OR 0.991; 95% IC 0.407-2.411; p=.983), when adjusted for procedure type, stone size and stone number. CONCLUSION: URS and RIRS can be done without fluoroscopic guidance in selected cases, without affecting the efficacy or safety of the procedure.

Distinct TNF-alpha and HLA polymorphisms associate with fibrotic and non-fibrotic subtypes of hypersensitivity pneumonitis.

INTRODUCTION: Since Hypersensitivity Pneumonitis (HP) categorization in fibrotic and nonfibrotic/inflammatory types seems to be more consistent with the distinctive clinical course and outcomes, recent international guidelines recommended the use of this classification. Moreover, fibrotic subtype may share immunogenetic and pathophysiological mechanisms with other fibrotic lung diseases. AIM: To investigate HLA -A, -B, -DRB1 and TNF-α -308 gene polymorphisms among fibrotic and nonfibrotic HP patients due to avian exposure, also in comparison with asymptomatic exposed controls. METHODS: We prospectively enrolled 40 HP patients, classified as fibrotic or nonfibrotic/inflammatory, and 70 exposed controls. HLA and TNF-α polymorphisms were determined by polymerase chain reaction-sequence specific primer amplification. RESULTS: While HLA alleles were not associated to HP susceptibility, fibrotic HP patients showed increased frequencies of HLA A*02 (46.7% vs 25.7%; OR=2.53, p = 0.02) and HLA DRB1*14 (10.0% vs 0.7%; OR=15.44, p=0.02) alleles when compared with exposed controls, although not statistically significant after correction for multiple comparisons. TNF-α G/G genotype (associated with low TNF-α production) frequencies were significantly increased among the non-fibrotic/inflammatory HP patients comparatively to fibrotic presentations (88% vs 60%; RR=0.44; p=0.04) and controls (88% vs 63%, OR 4.33, p=0.037). Also, these patients had a significantly increased frequency of the G allele (94.0% vs 73.3%, RR=0.44, p=0.01), while fibrotic HP patients predominantly presented the A allele (26.7% vs 6.0%, RR=2.28, p=0.01). CONCLUSIONS: Our results support the hypothesis that fibrotic and non-fibrotic HP subtypes exhibit a distinct profile of TNF-α and HLA polymorphisms, which may be relevant to predict disease course and better define treatment strategies.

Hypersensitivity pneumonitis: Main features characterization in a Portuguese cohort.

Hypersensitivity pneumonitis (HP) is an interstitial lung disease (ILD) which varies in prevalence across the world, depending on disease definition, diagnostic methods, exposure type and intensity, geographical environments, agricultural and industrial practices, and host risk factors. This study aimed to deepen knowledge about HP's clinical characteristics, diagnosis and functional and imaging features in a cohort of HP patients from the North of Portugal. To achieve this goal, a retrospective assessment of the clinical and diagnostic data was carried out, and patients were classified and compared according to disease presentation (acute, sub-acute and chronic HP forms). Of the 209 HP patients included (mean age 58.3 ±â€¯16.0 years), 52.6% were female and 73.7% presented a chronic form. Most patients had prior exposure to birds (76.6%). Dyspnoea and cough were the most frequently experienced symptoms, but no statistically significant differences were found between groups (p = 0.089, p = 0.418, respectively). Fever was most common in acute HP form (p < 0.001). The most common patterns found in Chest CT were ground glass (p = 0.002) in acute/subacute presentation, and reticulation (p < 0.001) in chronic form, while mosaic attenuation, although was also frequently observed, no statistically significant differences were found between groups (p = 0.512). The most common functional pattern was restrictive (38% of patients, 73.7% with chronic HP form). Bronchoalveolar lavage lymphocytes were higher in acute and subacute forms although not reaching statistical significance (p = 0.072), with lowest CD4/CD8 ratio (p = 0.001) in acute forms. Thus, given the significant disease heterogeneity, further studies with different populations and ambient exposures are needed to achieve a better stratification of the exposure risk, to provide proper implementation of avoidance methods and a precise diagnostic and therapeutic approach.

Hypersensitivity pneumonitis: Antigen diversity and disease implications.

Hypersensitivity pneumonitis (HP) is an immune-mediated syndrome triggered by inhalation of a wide variety of allergens, to which an individual has previously been sensitized. More than 200 agents responsible for the disease have already been identified; however, HP occurs only in a small number of individuals exposed to causal antigens. The present report provides an overview of the role of antigen role in HP, highlighting its diversity, research methods, and prevention strategies, as well as the impact on disease prognosis following elimination of antigen. HP is an underdiagnosed disease and, therefore, it is difficult to accurately estimate its incidence. Triggering antigens can be divided into six broad categories: bacteria, fungi, mycobacteria, animal and plant proteins, chemicals, and metals, represented by disease prototypes. The identification of causal antigen is a major challenge; it is impossible to obtain in about 30-60% of cases. The acute form of HP, with early detection and immediate eviction of causal antigen, tends to have an excellent prognosis. In the chronic form, partial recovery of disease is still possible; however, some cases tend to progress to fibrosis, even after removal from exposure. In conclusion, HP diagnosis should be based on a proactive search for potential antigen sources, although their identification is hampered by the lack of standardized methods of demonstrating the specific antigen sensitization. Antigen avoidance is a critical determinant in disease prognosis.

Predictive factors of obstructive sleep apnoea in patients with fibrotic lung diseases.

BACKGROUND AND AIM: Several studies reported a high prevalence of Obstructive Sleep Apnoea (OSA) in patients with Idiopathic Pulmonary Fibrosis (IPF) or restrictive end-stage lung disease (ESLD). Besides the known risk factors for OSA like high Body Mass Index (BMI), reduced static and dynamic volumes for IPF patients and reduced DLCO and low minimal O2 saturation during sleep for ESLD patients were associated with higher Apnoea-Hypopnoea Index (AHI). The aim of our study was to determine potential predictive factors of OSA in patients with Fibrotic Lung Diseases (FLD). MATERIALS AND METHODS: In this study, 49 patients with FLD and BMI ≤30 kg/m2 were included. All patients underwent portable cardiorespiratory polysomnography (PSG) and were asked to fill in Epworth Sleepiness Scale (ESS). Their epidemiological, medication and subsidiary exams data were retrieved from their hospital records. Univariate and multivariate correlation models were obtained. RESULTS: Approximately 70% of patients had an AHI ≥5 events/h. In an univariate correlation model, AHI showed a statistically significant correlation with age, BMI, the duration of immunosuppressant treatment, and Forced Expiratory Volume in the first second (FEV1). Only BMI remained an independent predictor of OSA in a multivariate correlation model adjusted for the other statistically meaningful variables. CONCLUSIONS: FLD patients, in general, show a prevalence of OSA superior to that of the general population. Excess of weight might predict a higher risk for OSA in FLD patients. Larger and more homogenous studies are warranted to clarify the associations between OSA severity and lung function impairment and the duration of immunosuppressant treatment.

Heterozygous TERT gene mutation associated with familial idiopathic pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease of unknown cause that occurs sporadically, but it can also occur in families and so named as Familial Pulmonary Fibrosis (FPF). Some forms of FPF overlaps IPF features, namely the radiological and histological pattern of usual interstitial pneumonia (UIP). Genetic and environmental factors commonly play an important role in the pathogenesis of FPF and the most commonly identified mutations involve the telomerase complex. Here, we report a rare case of FPF in a male at the age of 44, in whom genetic testing showed heterozygous variants for the telomerase reverse transcriptase gene (TERT). Our report highlights the importance of compiling a thorough family history in younger patients identified with UIP serving as a resource for identifying the current and future genetic links to disease. Families with UIP hold a great promise in defining UIP pathogenesis, potentially suggesting targets for the development of future therapies.

Organizing pneumonia revisited: insights and uncertainties from a series of 67 patients.

Background: Organizing pneumonia (OP) is classified as an acute/subacute pneumonia according to the American Thoracic Society/European Respiratory Society statement (2013 update). Although its clinical presentation, radiologic and histologic features are well established, data on the relevance of potential causes, corticosteroid doses and length, or management of relapses are based on heterogeneous series of patients. Objectives: The aims of this study were to describe clinical presentation, diagnosis and treatment of OP, explore potential causes, discuss strategies for managing relapses, and analyze prognostic factors. We also discuss our findings in relation to relevant data in the literature. Methods: We performed a cross-sectional study of all patients diagnosed with OP at a tertiary referral center in northern Portugal between 2008 and 2015. Results: Sixty-seven patients were diagnosed with OP over the 7-year study period. Dyspnea and cough were the most common presenting symptoms and approximately 30% of patients were hospitalized at the time of diagnosis. Approximately half of the patients were receiving drugs described as potential causes of OP. Microorganisms were isolated in approximately one-third of patients. Other potential causes identified were hematologic disorders, neoplasms, connective tissue diseases, myelodysplastic syndromes, immunodeficiencies, radiotherapy, and bird exposure. Cryptogenic OP was diagnosed in just 16 patients (23.8%). Corticosteroids were the most common treatment and 11 patients (16.4%) experienced relapse. Conclusions: The findings for this series of patients confirm the extreme variability of the contexts in which OP can occur and suggest that rather than a distinct, homogeneous clinicopathologic entity, OP is a non-specific reaction whose outcomes are dependent on the cause. (Sarcoidosis Vasc Diffuse Lung Dis 2018; 35: 129-138).

Effect of creatine on swimming velocity, body composition and hydrodynamic variables.

AIM: Creatine supplementation (CS) has been reported to increase body weight and improve performance during high intensity, short duration, exercise tasks. However, none of the published studies has investigated the influence of CS on performance related hydrodynamic variables during swimming. To investigate the effect of oral CS on swimming velocity, body composition and hydrodynamic variables during the period of final preparation of competitive junior female swimmers. METHODS: In a double blind and randomized manner, 16 female swimmers, were supplemented with 20 g day(-1) of creatine monohydrate (CS group), or a maltodextrin placebo (PL group) for 21 days. Just pre- and post-21 days of supplementation, subjects performed 2x25 swimming bouts at maximum velocity with a 3 min recovery between bouts. The variables measured were 25 m swimming velocity (MSV(25)); active drag force (D(f)); hydrodynamic coefficient (C(x)); power output (P(o)). Body measures were also analysed: body weight (kg), fat-mass (% FAT), body water (% H(2)O), and fat free mass (FFM). RESULTS: Significant differences were observed in hydrodynamic values: the CS group showed a significant reduction (approximately =25%), in D(f), C(x) and P(o) values, when comparing pretest with post-test. No differences were found in variables related to body composition and performance between CS group and PL group, as well as for CS group during the experimental period. CONCLUSIONS: These data suggest that 21 days of CS produced significant effects on gross and/or propelling efficiency during swimming in female athletes. However, CS did not influence performance, body weight and body composition.

Analysis of sarcoidosis in the Oporto region (Portugal).

BACKGROUND: Sarcoidosis is a systemic granulomatous disease of unknown etiology. Epidemiological studies of different populations are essential because clinical presentation, organ involvement, disease severity, and prognosis vary significantly according to region and population. The aim of this study was to assess epidemiological and clinical characteristics, staging factors, and clinical course in patients with sarcoidosis from a tertiary hospital in Oporto, Portugal. METHODS: A retrospective analysis of patients with sarcoidosis and at least 2 years of follow-up evaluated at the Centro Hospitalar de São João between 2000 and 2014. RESULTS: We identified 409 patients with sarcoidosis (females, 58.9%; mean age at diagnosis, 38.9±13.4 years; smokers, 14.4%]. All the patients were diagnosed according to the ERS/ATS/WASOG consensus statement and 64.1% had evidence of noncaseating epithelioid cell granulomas in biopsy specimens. Bronchoalveolar lavage was performed as part of the diagnostic work-up in 289 patients and 90.2% had lymphocytosis (CD4/CD8 ratio ≥3.5 in 60.9% of cases). Exertion dyspnea, cough, and constitutional symptoms were the most common presenting symptoms; 10.1% of patients were asymptomatic, 22.8% had Löfgren syndrome, and 50.5% had extrathoracic involvement. Radiographic stages of disease according to the Scadding criteria were as follows: stage 0 (5.2%), stage I (33.7%), stage II (47.0%), stage III (8.4%), and stage IV (5.7%). Impaired respiratory function was observed in 45.6% patients and was mostly mild. Systemic treatment was administered in 58.6% of cases. Overall, 45.3% of patients experienced disease resolution. CONCLUSION: The epidemiological and clinical characteristics of this cohort of patients with sarcoidosis from the Oporto region in northern Portugal revealed epidemiological and clinical characteristics that were generally similar to those described in other Western Europe populations and in the US ACCESS study. However, we found a higher proportion of patients who progressed to chronic forms.

Learning curve for transbronchial lung cryobiopsy in diffuse lung disease.

INTRODUCTION: Transbronchial lung cryobiopsy (TBLC) is increasingly used in the diagnosis of diffuse lung disease (DLD), but no data have yet been published on the learning curve associated with this technique. AIM: To evaluate diagnostic yield, lung tissue sample length and area, and procedure-related complications in a cohort of TBLC procedures to define the learning curve and threshold for proficiency. METHODS: Retrospective analysis of the first 100 TBLCs performed in different segments of the same lobe in patients with suspected DLD. We compared diagnostic yield, sample length and area, and complications between consecutive groups of patients. RESULTS: The overall diagnostic yield for TBLC was 82%. Median sample length was 5.4mm (IQR, 5-6) and median area was 19.5mm2 (IQR, 13.3-25). Pneumothorax was the most common complication (18%). On comparing the two groups of 50 consecutive patients, a significant difference was found for diagnostic yield (74% vs 90%; p=0.04), sample length (5.0mm [2.5-16] vs 6.0mm [4-12;] p<0.01) and area (17.5mm2 [6-42] vs 21.5mm2 [10-49]; p<0.01). Logarithm regression was applied to median diagnostic yield and sample length and area for groups of 10 consecutive patients to define the learning curve, which plateaued after approximately 70 procedures. CONCLUSIONS: Our findings suggest that proficiency in TBLC is achieved at approximately the 70th procedure; however they need to be validated in more series and cohorts.

Spermatogonial stem cell organization in felid testis as revealed by Dolichos biflorus lectin.

Spermatogonial stem cells are being exploited in many species as a tool to recover fertility, but may also be used to manipulate the genetic pool. Whatever the purpose, these cells must be fully characterized and easily identifiable, and our goal was to improve this procedure in the domestic cat, used as an animal model for endangered felid species and for some human diseases/physiological processes. We have therefore screened several markers that might be used to distinguish and study the undifferentiated spermatogonia population in situ and in vitro via immunohistochemistry applied to tissue sections and whole mounts of the domestic cat seminiferous tubules. Our results show that, although they label the cytoplasm and nucleus of gonocytes and spermatogonia in pre-pubertal animals, PGP9.5 and FoxO1 cannot be considered markers of undifferentiated spermatogonia in adult animals, as almost all spermatogonia, namely type A and B, express these proteins. Nonetheless, the Dolichos biflorus agglutinin (DBA lectin) was able to label the cell surface and cytoplasm of a small type A spermatogonial population in the adult animals. Analysis of the number and distribution of the DBA-labeled cells showed they were present in low number, which did not vary with epithelium seminiferous stage. Morphometric analysis revealed that DBA-labeled cells present tropism to a peculiar area of the seminiferous tubules, namely the area in direct contact with Leydig cells. Whole mounts of DBA-stained seminiferous tubules revealed the arrangement of DBA-stained cells in small clones up to eight cells. Noteworthy, the clonal cells presented variable staining intensity suggesting the existence of asymmetric distribution of O-glycosylated proteins within each clone. Our results strongly suggest that the DBA lectin is a marker of undifferentiated spermatogonia in domestic cat, and illustrate the peculiar characteristics of spermatogonial stem cell development and organization in this species.

Tuberculosis Caused by Mycobacterium bovis in a Capybara (Hydrochoerus hydrochaeris).

Tuberculosis, associated with Mycobacterium bovis, was diagnosed post mortem in an adult female capybara (Hydrochoerus hydrochaeris), kept at the Pampulha Ecological Park, Belo Horizonte, Brazil, in a large metropolitan area. On post-mortem examination, there were numerous firm white nodules scattered throughout all lobes of both lungs. Tissue samples were collected for histological and microbiological examination. Microscopically, the pulmonary nodules were multifocal to coalescing granulomas and intralesional acid-fast bacilli were evident in Ziehl-Neelsen-stained sections of the lung and spleen. Colonies with morphological features of Mycobacterium spp. were isolated from lung samples and conventional polymerase chain reaction (PCR) with genomic DNA from the isolates was positive for M. bovis; sequencing indicated 100% identity with the region of difference 4 (RD4) of M. bovis. In addition, M. bovis DNA was detected in the lung by quantitative PCR. The finding of M. bovis in a capybara indicates a potential public health risk in a zoological collection.