Modulation of small leucine-rich proteoglycans (SLRPs) expression in the mouse uterus by estradiol and progesterone

<p>Abstract</p> <p>Background</p> <p>We have previously demonstrated that four members of the family of small leucine-rich-proteoglycans (SLRPs) of the extracellular matrix (ECM), named decorin, biglycan, lumican and fibromodulin, are deeply remodeled in mouse uterine tissues along the estrous cycle and early pregnancy. It is known that the combined action of estrogen (E2) and progesterone (P4) orchestrates the estrous cycle and prepares the endometrium for pregnancy, modulating synthesis, deposition and degradation of various molecules. Indeed, we showed that versican, another proteoglycan of the ECM, is under hormonal control in the uterine tissues.</p> <p>Methods</p> <p>E2 and/or medroxiprogesterone acetate (MPA) were used to demonstrate, by real time PCR and immunoperoxidase staining, respectively, their effects on mRNA expression and protein deposition of these SLRPs, in the uterine tissues.</p> <p>Results</p> <p>Decorin and lumican were constitutively expressed and deposited in the ECM in the absence of the ovarian hormones, whereas deposition of biglycan and fibromodulin were abolished from the uterine ECM in the non-treated group. Interestingly, ovariectomy promoted an increase in decorin, lumican and fibromodulin mRNA levels, while biglycan mRNA conspicuously decreased. Hormone replacement with E2 and/or MPA differentially modulates their expression and deposition.</p> <p>Conclusions</p> <p>The patterns of expression of these SLRPs in the uterine tissues were found to be hormone-dependent and uterine compartment-related. These results reinforce the existence of subpopulations of endometrial fibroblasts, localized into distinct functional uterine compartments, resembling the organization into basal and functional layers of the human endometrium.</p

Hormone-regulated expression and distribution of versican in mouse uterine tissues

<p>Abstract</p> <p>Background</p> <p>Remodeling of the extracellular matrix is one of the most striking features observed in the uterus during the estrous cycle and after hormone replacement. Versican (VER) is a hyaluronan-binding proteoglycan that undergoes RNA alternative splicing, generating four distinct isoforms. This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination.</p> <p>Methods</p> <p>Uteri from mice in all phases of the estrous cycle, and animals subjected to ovariectomy and hormone replacement were collected for immunoperoxidase staining for versican, as well as PCR and quantitative Real Time PCR.</p> <p>Results</p> <p>In diestrus and proestrus, VER was exclusively expressed in the endometrial stroma. In estrus and metaestrus, VER was present in both endometrial stroma and myometrium. In OVX mice, VER immunoreaction was abolished in all uterine tissues. VER expression was restored by E2, MPA and E2+MPA treatments. Real Time PCR analysis showed that VER expression increases considerably in the MPA-treated group. Analysis of mRNA identified isoforms V0, V1 and V3 in the mouse uterus.</p> <p>Conclusion</p> <p>These results show that the expression of versican in uterine tissues is modulated by ovarian steroid hormones, in a tissue-specific manner. VER is induced in the myometrium exclusively by E2, whereas MPA induces VER deposition only in the endometrial stroma.</p

Estradiol induces transcriptional and posttranscriptional modifications in versican expression in the mouse uterus

We have previously shown the differential expression of versican in the mouse uterus under ovarian hormone influence. We also demonstrated there is not a direct correlation between mRNA levels and protein expression, suggesting posttranscriptional events, such as alteration in mRNA stability. This posttranscriptional effect may result in the elongation and stabilization of transcripts poly(A) tail. Thus, the aim of this study was to analyze whether estradiol (E2) regulates versican mRNA stability and expression in a dose-related and time-dependent manner. For this purpose female mice were ovariectomized and treated with a single injection of 0.1 or 10 μg E2. To block transcription a group of females received a single injection of alpha-amanitin before hormone administration. Uterine tissues were collected 30 min, 1, 3, 6, 12 and 24 h after treatments and processed for quantitative real time PCR (qPCR), RACE-PAT Assay and immunohistochemistry. qPCR showed that versican mRNA levels are higher than control from 3 to 24 h after E2 administration, whereas after transcription inhibition versican mRNA unexpectedly increases within 3 h, which can be explained when transcriptional blockers alter the degradation rate of the transcript, resulting in the superinduction of this mRNA. Accordingly, analysis of versican transcript poly(A) tail evidenced a longer product 3 h after treatment, but not after 12 h. Versican immunoreaction becomes conspicuous in the superficial stroma only 3 h after E2 injection, whereas the whole stroma is immunoreactive from 6 h onward. These results demonstrate that E2 modulates versican at the transcriptional and posttranscriptional levels in a time-dependent manner.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) - Brazil (09/51788-1)CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior)CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico

Effect of photodynamic therapy with malachite green on non-surgical periodontal treatment in HIV patients : a pilot split-mouth study

A number of oral lesions are among the early features of HIV infection. It has been described that HIV patients are at risk for severe periodontal diseases. In addition, there is a higher prevalence of periodontal pathogens such as Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Tannerella forsythensis (Tf) in HIV patients compared with non-HIV patients. Polymerase chain reaction (PCR) is a diagnostic method that enables assessing microorganisms with very high levels of accuracy. For this reason, PCR is helpful to detect periodontal pathogens at levels below the ones detectable by cell culture or other diagnostic methods. In most cases, the sole use of repeated scaling and root planing (SRP) as periodontal therapy leads to satisfactory clinical outcomes. However, the aforementioned approach may not be enough to achieve periodontal health in more challenging cases, such as residual deep pockets and furcation areas . In addition, SRP often needs to be followed by adjunctive therapy such as local delivery and systemic antimicrobials and host modulation. One of the most recent methods used in combination with SRP in the periodontal treatment is the photodynamic therapy (PDT). In PDT, a photosensitizing agent is used to apply light therapy selectively to target specific cells. It is useful for sensitizing bacterial cells, leading to effective antimicrobial activity due to the production of cytotoxic oxygen free radicals (singlet oxygen). However, the efficacy of PDT is dependent on various factors, such as the laser wavelength and its interaction with the photosensitizer. One of the photosensitizers that have been used for the aforementioned purpose is the malachite green (MG), which leads to dissipation of the cell membrane potential in both gram-positive and - negative bacterial species. MG is defined as a cationic dye of the triarylmethane family that shows satisfactory absorption at the red end of the visible spectrum. However, little is known on the effect of PDT with MG as an adjunctive periodontal treatment in combination with SRP. Thus, the aim of this pilot split-mouth study was to assess clinical and microbiological effects of PDT with MG on non-surgical periodontal treatment in HIV patients.peer-reviewe

Long-term effects of competition and environmental drivers on the growth of the endangered coral Mussismilia braziliensis (Verril, 1867)

Most coral reefs have recently experienced acute changes in benthic community structure, generally involving dominance shifts from slow-growing hard corals to fast-growing benthic invertebrates and fleshy photosynthesizers. Besides overfishing, increased nutrification and sedimentation are important drivers of this process, which is well documented at landscape scales in the Caribbean and in the Indo-Pacific. However, small-scale processes that occur at the level of individual organisms remain poorly explored. In addition, the generality of coral reef decline models still needs to be verified on the vast realm of turbid-zone reefs. Here, we documented the outcome of interactions between an endangered Brazilian-endemic coral (Mussismilia braziliensis) and its most abundant contacting organisms (turf, cyanobacteria, corals, crustose coralline algae and foliose macroalgae). Our study was based on a long (2006–2016) series of high resolution data (fixed photoquadrats) acquired along a cross-shelf gradient that includes coastal unprotected reefs and offshore protected sites. The study region (Abrolhos Bank) comprises the largest and richest coralline complex in the South Atlantic, and a foremost example of a turbid-zone reef system with low diversity and expressive coral cover. Coral growth was significantly different between reefs. Coral-algae contacts predominated inshore, while cyanobacteria and turf contacts dominated offshore. An overall trend in positive coral growth was detected from 2009 onward in the inshore reef, whereas retraction in live coral tissue was observed offshore during this period. Turbidity (+) and cyanobacteria (−) were the best predictors of coral growth. Complimentary incubation experiments, in which treatments of Symbiodinium spp. from M. braziliensis colonies were subjected to cyanobacterial exudates, showed a negative effect of the exudate on the symbionts, demonstrating that cyanobacteria play an important role in coral tissue necrosis. Negative effects of cyanobacteria on living coral tissue may remain undetected from percent cover estimates gathered at larger spatial scales, as these ephemeral organisms tend to be rapidly replaced by longer-living macroalgae, or complex turf-like consortia. The cross-shelf trend of decreasing turbidity and macroalgae abundance suggests either a direct positive effect of turbidity on coral growth, or an indirect effect related to the higher inshore cover of foliose macroalgae, constraining cyanobacterial abundance. It is unclear whether the higher inshore macroalgal abundance (10–20% of reef cover) is a stable phase related to a long-standing high turbidity background, or a contemporary response to anthropogenic stress. Our results challenge the idea that high macroalgal cover is always associated with compromised coral health, as the baselines for turbid zone reefs may derive sharply from those of coral-dominated reefs that dwell under oligotrophic conditions

A different immunologic profile characterizes patients with HER-2-overexpressing and HER-2-negative locally advanced breast cancer: implications for immune-based therapies

INTRODUCTION: The clinical efficacy of trastuzumab and taxanes is at least partly related to their ability to mediate or promote antitumor immune responses. On these grounds, a careful analysis of basal immune profile may be capital to dissect the heterogeneity of clinical responses to these drugs in patients with locally advanced breast cancer undergoing neoadjuvant chemotherapy. METHODS: Blood samples were collected from 61 locally advanced breast cancers (36 HER2- and 25 HER2+) at diagnosis and from 23 healthy women. Immunophenotypic profiling of circulating and intratumor immune cells, including regulatory T (Treg) cells, was assessed by flow cytometry and immunohistochemistry, respectively. Serum levels of 10 different cytokines were assessed by multiplex immunoassays. CD8+ T cell responses to multiple tumor-associated antigens (TAA) were evaluated by IFN-γ-enzyme-linked immunosorbent spot (ELISPOT). The Student's t test for two tailed distributions and the Wilcoxon two-sample test were used for the statistical analysis of the data. RESULTS: The proportion of circulating immune effectors was similar in HER2+ patients and healthy donors, whereas higher percentages of natural killer and Treg cells and a lower CD4+/CD8+ T cell ratio (with a prevalence of naïve and central memory CD8+ T cells) were observed in HER2- cases. Higher numbers of circulating CD8+ T cells specific for several HLA-A*0201-restricted TAA-derived peptides were observed in HER2+ cases, together with a higher prevalence of intratumor CD8+ T cells. Serum cytokine profile of HER2+ patients was similar to that of controls, whereas HER2- cases showed significantly lower cytokine amounts compared to healthy women (IL-2, IL-8, IL-6) and HER2+ cases (IL-2, IL-1β, IL-8, IL-6, IL-10). CONCLUSIONS: Compared to HER2- cases, patients with HER2-overexpressing locally advanced breast cancer show a more limited tumor-related immune suppression. This may account for the clinical benefit achieved in this subset of patients with the use of drugs acting through, but also promoting, immune-mediated effects

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure &lt;= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost