Plasmid pB1000 is a mobilizable replicon bearing the blaROB-1 ss-lactamase gene that we have recently described in Haemophilus parasuis and Pasteurella multocida animal isolates. Here, we report presence of pB1000 and a derivate plasmid, pB1000', in four H. influenzae clinical isolates from human origin. Pulsed-field gel electrophoresis showed unrelated patterns in all strains, indicating that existence of pB1000 in H. influenzae isolates is not the consequence of clonal dissemination. The replicon can be transferred both by transformation and conjugation into H. influenzae, giving rise to ampicillin (MIC > 64 mug/ml) and cefaclor (MIC > 64 mug/ml) resistant recipients. Stability experiments showed that pB1000 is stable in H. influenzae without antimicrobial pressure for at least 60 generations. Competition experiments between isogenic H. influenzae with and without pB1000 revealed a competitive disadvantage of 9% per 10 generations of the transformant versus the recipient. Complete nucleotide sequence of nine pB1000 plasmids from human and animal isolates, as well as the epidemiological data, suggest that animal Pasteurellaceae act as antimicrobial resistance reservoir for H. influenzae. Further, being P. multocida the only member of this family able to colonize both, humans and animals, we propose P. multocida as the vehicle of pB1000 between animal- and human-adapted Pasteurellaceae.

Banff classification empirically established scoring of histologic lesions, but the relationships of lesions to each other and to underlying biologic processes remain unclear. We hypothesized that class discovery tools would reveal new relationships between individual lesions, and relate lesions to C4d staining, anti-HLA donor-specific antibody (DSA) and time posttransplant. We studied 234 nonselected renal allograft biopsies for clinical indications from 173 patients. Silhouette plotting and principal component analysis revealed three groups of lesions: microcirculation changes, including inflammation (glomerulitis, capillaritis) and deterioration (double contours, mesangial expansion); scarring/hyalinosis; and tubulointerstitial inflammation. DSA and C4d grouped with microcirculation inflammation, whereas time posttransplant grouped with scarring/hyalinosis lesions. Intimal arteritis clustered with DSA, C4d and microcirculation inflammation, but also showed correlations with tubulitis. Fibrous intimal thickening in arteries clustered with scarring/hyalinosis. Capillary basement membrane multilayering showed intermediary relationships between microcirculation deterioration and time-dependent scarring. Correlation analysis and hierarchical clustering confirmed the lesion relationships. Thus, we propose that the pathologic lesions in biopsies are not independent but are members of groups that represent distinct pathogenic forces: microcirculation changes, reflecting the stress of DSA; scarring, hyalinosis and arterial fibrosis, reflecting the cumulative burden of injury over time; and tubulointerstitial inflammation. Interpretation of lesions should reflect these associations.

BACKGROUND: Several studies drawn from the Ecuadorian population have previously reported that more than half of mid-aged women present hot flushes, which can impair their quality of life. However up-to-date risk factors for their presence and severity have not been assessed. OBJECTIVE: To assess hot flush frequency and intensity and related risk factors among middle-aged Ecuadorian women. METHODS: In this cross-sectional study, 1154 healthy women aged 40-59 years, visiting healthcare centers of eight main cities of Ecuador with more than 100,000 inhabitants, were assessed with the first item of the Menopause Rating Scale (MRS) and a questionnaire containing female and partner socio-demographic data. RESULTS: Mean age of the entire sample was 48.8+/-5.6 years (median 48), a 48.7% had 12 or less years of schooling, 52.8% were postmenopausal, 43.6% lived at high altitude, 56.8% were married and 10% were on hormonal therapy (HT). Hot flushes accounted for 56%(n=646) of the whole sample, of which 29.1% and 9.1% were respectively graded as severe and very severe. Logistic regression determined that female sedentarism (OR: 2.42, CI 95%[1.63-3.59]), accessing a free healthcare system (OR: 1.96, CI 95%[1.30-2.96]), living at high altitude (OR: 1.82, CI 95%[1.14-2.90]) and having a partner abusing alcohol (OR: 1.92, CI 95%[1.09-3.35]) were significant risk factors related to the presence of hot flushes. The regression model also determined that among women with hot flushes (n=646), sedentarism (OR: 1.73, CI 95%[1.14-2.62]) and having a partner with erectile dysfunction (OR: 2.57, CI 95%[1.44-4.59]) were significant risk factors related to severe/very severe hot flushes whereas married status (OR: 0.53, CI 95%[0.32-0.86]), living at high altitude (OR: 0.46, CI 95%[0.26-0.78]) and partner healthiness (OR: 0.59, CI 95%[0.36-0.95]) were not. CONCLUSION: To the best of our knowledge this is the first and largest study assessing hot flushes in a mid-aged Ecuadorian population. We found that the presence and severity were not significantly related to age and hormonal status yet to other individual female/male characteristics and the demography of the studied population.

Increasing evidence includes Wnt proteins inside the group of master-signaling pathways that govern immune and nonimmune differentiation systems, fundamental for normal development and homeostasis. Although their precise functions in bone marrow and thymus are still controversial, numerous studies have shown that Wnt signaling is able to control the proliferation of hematopoietic stem cells and thymic progenitors and might also affect their cell-fate decisions and subsequent maturation. In the present work, we analyze the effect of transient stimulation of the canonical Wnt pathway in the differentiation potential of Lin(-)CD34(+) CD1a(-) human thymic progenitors, a multipotent and heterogeneous cell population that has the capacity to develop into T cells, NK cells, monocytes, cDC, and pDC. Our results demonstrate that giving a boost to canonical Wnt signaling, triggered by transient exposure to Wnt3a or LiCl, the differentiation capacity of thymic progenitors changes, enhancing NK cell production. On the contrary, Wnt3a- or LiCl-pretreated thymic progenitors generate a significantly lower number of myeloid lineage cells, monocytes, and cDC and exhibit a reduced capacity to differentiate into pDC lineage. As a possible mechanism for this effect, we show that Wnt3a- and LiCl-pretreated progenitors change their membrane levels of receptors for cytokines pivotal for their expansion and differentiation, such as Flt3L. Moreover, canonical Wnt pathway stimulation modifies the transcription factor profile of CD34(+)CD1(-) thymocytes, increasing Hes-1 and ID3 expression levels.

BACKGROUND: Sleep disorders and sleep-apnea/hypopnea syndromes are very frequent in women, being misdiagnosed in many cases. The menopause, regardless of age, is associated to poor sleep quality and daytime sleepiness that can lead to impaired quality of life, and reduced productivity and functioning. OBJECTIVE: To assess daytime sleepiness and related risk factors among middle aged Ecuadorian women using the Epworth Sleepiness Scale (ESS). METHODS: In this cross-sectional study 149 women aged 40-59 years were assessed for hot flush presence and intensity using the Menopause Rating Scale (MRS) and requested to fill out the ESS and a questionnaire containing personal and partner data. RESULTS: Mean age of surveyed women was 47.6+/-5.5 years, with 67.8% having less than 12 years of schooling, 33.6% being postmenopausal, and 2.7% on hormone therapy. A 10.1% were current smokers and 20.8% were sedentary. According to the MRS (item 1) 51.7% presented hot flushes, which were graded as severe-very severe in 42.8% of cases. Regarding the partner (n=132), erectile dysfunction was present in 10.6%, premature ejaculation 6.1% and 17.4% abused alcohol. Mean total ESS score was 8+/-4.4 (median 8), with 33.6% considered having some degree of daytime sleepiness (ESS score >/=10). Logistic regression analysis determined that postmenopausal status (OR 6.58, CI 95%[2.51-17.23], p=0.001), sedentarism (OR 3.43, CI 95%[1.14-10.26], p=0.02) and hot flush presence (OR 2.61, CI 95%[1.02-6.65], p=0.04) among women were risk factors for increased daytime sleepiness (ESS total score >/=10) whereas partner faithfulness decreased this risk (OR 0.47, CI 95%[0.24-0.90], p=0.02). CONCLUSION: Increased daytime sleepiness in this middle aged series was related to female (hormonal status and sedentarism) and partner factors; several which are susceptible of intervention.

Bone morphogenetic proteins (BMPs) play a pivotal role during vertebrate embryogenesis and organogenesis, and have also been described to function in regulating cell fate and determination in self-renewing tissues in adults. Recent results have demonstrated that the different components of the BMP2/4 signaling pathway are expressed in the human thymus. In this study, we provide evidence that BMP4 and IL-7 interplay is important in the maintenance of the human thymic progenitor population. Intrathymic CD34(+) cells express BMP receptors (BMPRIA, BMPRIB, ActRIA, BMPRII), signal transduction molecules (Smad1, 5, 8 and 4), and produce BMP4. Neutralization of endogenous BMP4 by treatment with the antagonist Noggin reduces thymic precursor cell survival, and the addition of exogenous BMP4 decreases their proliferation. The treatment of chimeric human-mouse fetal thymus organ cultures with BMP4 inhibits cell expansion, arrests thymocyte differentiation, and leads to the accumulation of human CD34(+) precursor cells. This effect is mainly attributed to the ability of BMP4 to counteract the IL-7-induced proliferation and differentiation of CD34(+) cells. BMP4 downregulates in the precursor cell population the expression of CD127 and inhibits the IL-7-dependent STAT5 phosphorylation. In addition, BMP signaling is promoted by IL-7. Our results also demonstrate that in thymic progenitors BMPs act downstream of Sonic Hedgehog, previously described to function as a maintenance factor for human intrathymic CD34(+) precursor cells.

Background. Postmenopausal metabolic changes increase cardiovascular risk and impair quality of life (QoL). Despite this, few reports have addressed the association of these changes with female sexuality. Objective. To determine the association between the metabolic syndrome (METS), and its components, and female sexuality. Methods. Data of sexually active postmenopausal women who participated in a METS screening program who filled out the menopause-specific quality of life questionnaire (MENQOL) were assessed. Specifically the sexual domain of the MENQOL was analyzed in regard to mean total and item scores (decreased libido, vaginal dryness, and sexual avoidance). Criteria of the Third Adult Treatment Panel (ATP III) were used to identify women with the METS. Results. Two hundred six women fulfilled inclusion criteria. Mean age of participants was 54 +/- 6.9 years (median: 54 years). Prevalence of the METS in this sexually active postmenopausal series was 39.8%. About 52.9% of them presented abdominal obesity, 35.4% hypertension, 55.8% high triglycerides, 17.5% hyperglycemia, and 59.7% decreased high density lipoprotein cholesterol (HDL-C). Women with the METS as compared with those without the syndrome displayed no significant differences in MENQOL sexual scorings (total or of its composing items). Equally there were also no score differences among those presenting any of the five components of the METS, except women with hyperglycemia who significantly displayed a higher total sexual domain score (5.6 +/- 2.1 vs. 4.8 +/- 2.3, p < 0.05) in association to a higher mean score in the decreased libido item (6.0 +/- 2.3 vs. 4.8 +/- 2.6, p < 0.01). After controlling for several confounding factors, logistic regression confirmed that women with hyperglycemia were significantly at higher risk for presenting decreased libido (higher item score, OR 2.4, CI 95%: 1.0-5.7, p < 0.05) and more impaired sexuality (higher total MENQOL sexual domain score: OR, 2.5, CI 95%: 1.1-5.4, p < 0.05). Conclusion. Despite the limitations of this study, as assessed with the MENQOL, hyperglycemia in postmenopausal women screened for the METS was associated to a negative impact in sexuality. More research is warranted in this regard.

We studied whether de novo donor-specific antibodies (DSA) in sera from patients undergoing kidney transplant biopsies associate with specific histologic lesions in the biopsy and prognosis. DSA were assessed in 145 patients at the time of biopsy between 7 days to 31 years posttransplant. DSA was detected in 54 patients (37%), of which 32 represented de novo DSA. De novo DSA was more frequent in patients having late biopsies (34%) versus early biopsies (4%), and was usually either against class II alone or class I and II but rarely against class I alone. Microcirculation inflammation (glomerulitis, capillaritis) and damage (glomuerulopathy, capillary basement membrane multilayering), and C4d staining were associated with de novo DSA. However, the degree of scarring, arterial fibrosis and tubulo-interstitial inflammation did not correlate with the presence of de novo DSA. De novo DSA correlated with reduced graft survival after the biopsy. Thus, de novo DSA at the time of a late biopsy for clinical indication is primarily against class II, and associates with microcirculation changes in the biopsy and subsequent graft failure. We propose careful assessment of de novo DSA, particularly against class II, be performed in all late kidney transplant biopsies.

We studied the phenotype of late kidney graft failure in a prospective study of unselected kidney transplant biopsies taken for clinical indications. We analyzed histopathology, HLA antibodies and death-censored graft survival in 234 consecutive biopsies from 173 patients, taken 6 days to 31 years posttransplant. Patients with late biopsies (>1 year) frequently displayed donor-specific HLA antibody (particularly class II) and microcirculation changes, including glomerulitis, glomerulopathy, capillaritis, capillary multilayering and C4d staining. Grafts biopsied early rarely failed (1/68), whereas grafts biopsied late often progressed to failure (27/105) within 3 years. T-cell-mediated rejection and its lesions were not associated with an increased risk of failure after biopsy. In multivariable analysis, graft failure correlated with microcirculation inflammation and scarring, but C4d staining was not significant. When microcirculation changes and HLA antibody were used to define antibody-mediated rejection, 17/27 (63%) of late kidney failures after biopsy were attributable to antibody-mediated rejection, but many were C4d negative and missed by current diagnostic criteria. Glomerulonephritis accounted for 6/27 late losses, whereas T-cell-mediated rejection, drug toxicity and unexplained scarring were uncommon. The major cause of late kidney transplant failure is antibody-mediated microcirculation injury, but detection of this phenotype requires new diagnostic criteria.