Influence of sex on chronic steroid-induced glaucoma: 24-Weeks follow-up study in rats.

The objective was to evaluate ocular changes based on sex in steroid-induced glaucoma models in rats comparing healthy controls, over 24 weeks follow-up. Eighty-nine Long-Evans rats (38 males and 51 females) with steroid-induced glaucoma were analysed. Two steroid-induced glaucoma models were generated by injecting poly-co-lactic-glycolic acid microspheres loaded with dexamethasone (MMDEX model) and dexamethasone-fibronectin (MMDEXAFIBRO model) into the ocular anterior chamber. Intraocular pressure was measured by rebound tonometer Tonolab®. Neuroretinal function was analysed using dark- and light-adapted electroretinography (Roland consult® RETIanimal ERG), and structure was analysed using optical coherence tomography (OCT Spectralis, Heidelberg® Engineering) using Retina Posterior Pole, Retinal Nerve Fibre Layer and Ganglion Cell Layer protocols over 24 weeks. Males showed statistically (p < 0.05) higher intraocular pressure measurements. In both sexes and models neuroretinal thickness tended to decrease over time. In the MMDEX model, males showed higher IOP values and greatest percentage thickness loss in the Ganglion Cell Layer (p = 0.015). Females receiving MMDEXAFIBRO experienced large fluctuations in thickness, a higher percentage loss (on average) in Retina Posterior Pole (p = 0.035), Retinal Nerve Fibre Layer and Ganglion Cell Layer than aged-matched males, and the highest thickness loss rate by mmHg. Although no difference was found by sex in dark- and light-adapted electroretinography, increased amplitude in photopic negative response was found in MMDEX males and MMDEXAFIBRO females at 12 weeks. Although both glaucoma models used dexamethasone, different intraocular pressure and neuroretinal changes were observed depending on sex and other influential cofactors (fibronectin). Both sex and the induced glaucoma model influenced neuroretinal degeneration.

Deroofing: A safe, effective and well-tolerated procedure in patients with hidradenitis suppurativa.

INTRODUCTION: Hidradenitis suppurativa (HS) is a recurrent, debilitating, chronic disorder of the pilosebaceous unit. Although advances in HS treatment have been made, more than 45% of patients remain dissatisfied with systemic treatment, and more than one-third are dissatisfied with surgical procedures. OBJECTIVES: A prospective, observational study on the deroofing procedures in HS with special attention paid to patient satisfaction and complications. METHODS: HS lesions were assessed clinically and by the use of ultrasound. Patients reported outcomes, including pain, itch and satisfaction, were measured at 24 h post-surgery by a numeric rating scale (NRS) ranging from 0 to 10. Additionally, the timeline of objective wound closure reported by patients in (weeks), in addition to the need for any analgesics use, were both evaluated. RESULTS: The mean closure time of the post-deroofing wound was assessed as 4.4 ± 1.9 weeks. A statistically longer time was necessary for complete closure in males than in females (4.9 ± 2.2 weeks and 3.9 ± 1.6 weeks, respectively; p = 0.046). The closure time correlated positively yet weakly with the HS tunnel's width (r = 0.27, p = 0.016) and length (r = 0.228, p = 0.044). Patients assessed mean pain at 24 h post-op as mild with 0.7 ± 1.2 points according to NRS, with no differences between sexes. Similarly, itch in the first 24 h was assessed as mild with 1.8 ± 1.1 points, without differences between sexes. No pain, itch or adverse events were reported after 1 week following deroofing. Moreover, no cases of wound infection were reported. An overall patient satisfaction was assessed as 9.9 ± 0.4 points (range 9-10 points). CONCLUSION: Deroofing is an easy, effective and safe dermatosurgical procedure that does not require surgical experience or operating theatre. It is associated with no complications and very low post-op pain and should be part of holistic HS management.

Toward Precision Medicine in Atopic Dermatitis Using Molecular-Based Approaches.

Atopic dermatitis is the most common chronic inflammatory skin disorder, affecting up to 20% of children and 10% of adults in developed countries. The pathophysiology of atopic dermatitis is complex and involves a strong genetic predisposition and T-cell driven inflammation. Although our understanding of the pathology and drivers of this disease has improved in recent years, there are still knowledge gaps in the immune pathways involved. Therefore, advances in new omics technologies in atopic dermatitis will play a key role in understanding the pathogenesis of this burden disease and could develop preventive strategies and personalized treatment strategies. In this review, we discuss the latest developments in genetics, transcriptomics, epigenomics, proteomics, and metagenomics and understand how integrating multiple omics datasets will identify potential biomarkers and uncover nets of associations between several molecular levels.

Toward Precision Medicine in Atopic Dermatitis Using Molecular-Based Approaches. / [Artículo traducido] Hacia una medicina de precisión en la dermatitis atópica mediante el uso de enfoques moleculares.

Atopic dermatitis is the most common chronic inflammatory skin disorder, affecting up to 20% of children and 10% of adults in developed countries. The pathophysiology of atopic dermatitis is complex and involves a strong genetic predisposition and T-cell driven inflammation. Although our understanding of the pathology and drivers of this disease has improved in recent years, there are still knowledge gaps in the immune pathways involved. Therefore, advances in new omics technologies in atopic dermatitis will play a key role in understanding the pathogenesis of this burden disease and could develop preventive strategies and personalized treatment strategies. In this review, we discuss the latest developments in genetics, transcriptomics, epigenomics, proteomics, and metagenomics and understand how integrating multiple omics datasets will identify potential biomarkers and uncover nets of associations between several molecular levels.

Acute fetal cardiovascular adaptation to artificial placenta in sheep model.

OBJECTIVE: To describe the acute cardiovascular adaptation of the fetus after connection to an artificial placenta (AP) in a sheep model, using ultrasound and invasive and non-invasive hemodynamic assessment. METHODS: This was an experimental study of 12 fetal sheep that were transferred to an AP system, consisting of a pumpless circuit with umbilical cord connection, at 109-117 days' gestation. The study was designed to collect in-utero and postcannulation measurements in all the animals. The first six consecutive fetuses were fitted with intravascular catheters and perivascular probes to obtain invasive physiological data, including arterial and venous intravascular pressures and perivascular blood flows, with measurements taken in utero and at 5 and 30 min after cannulation. These experiments were designed with a survival goal of 1-3 h. The second set of six fetuses were not fitted with catheters, and experiments were aimed at 3-24 h of survival. Echocardiographic assessment of cardiac anatomy and function, as well as measurements of blood flow and pre- and postmembrane pressures recorded by circuit sensors in the AP system, were available for most of the fetuses. These data were acquired in utero and at 30 and 180 min after cannulation. RESULTS: Compared with in-utero conditions, the pulsatility index at 30 and 180 min after connection to the AP system was reduced in the umbilical artery (median, 1.36 (interquartile range (IQR), 1.06-1.50) vs 0.38 (IQR, 0.31-0.50) vs 0.36 (IQR, 0.29-0.41); P < 0.001 for extreme timepoints) and the ductus venosus (median, 0.50 (IQR, 0.41-0.67) vs 0.29 (IQR, 0.22-0.33) vs 0.36 (IQR, 0.22-0.41); P = 0.011 for extreme timepoints), whereas umbilical venous peak velocity increased (median, 20 cm/s (IQR, 18-22 cm/s) vs 39 cm/s (IQR, 31-43 cm/s) vs 43 cm/s (IQR, 34-54 cm/s); P < 0.001 for extreme timepoints) and flow became more pulsatile. Intravascular monitoring showed that arterial and venous pressures increased transiently after connection, with median values for mean arterial pressure at baseline, 5 min and 30 min of 43 mmHg (IQR, 35-54 mmHg), 72 mmHg (IQR, 61-77 mmHg) and 58 mmHg (IQR, 50-64 mmHg), respectively (P = 0.02 for baseline vs 5 min). Echocardiography showed a similar transient elevation of fetal heart rate at 30 and 180 min after connection compared with in utero (median, 145 bpm (IQR, 142-156 bpm) vs 188 bpm (IQR, 171-209 bpm) vs 175 bpm (IQR, 165-190 bpm); P = 0.001 for extreme timepoints). Fetal cardiac structure and function were mainly preserved; median values for right fractional area change were 36% (IQR, 34-41%) in utero, 38% (IQR, 30-40%) at 30 min and 37% (IQR, 33-40%) at 180 min (P = 0.807 for extreme timepoints). CONCLUSIONS: Connection to an AP system resulted in a transient fetal hemodynamic response that tended to normalize over hours. In this short-term evaluation, cardiac structure and function were preserved. However, the system resulted in non-physiologically elevated venous pressure and pulsatile flow, which should be corrected to avoid later impairment of cardiac function. © 2023 International Society of Ultrasound in Obstetrics and Gynecology.

Spatial distribution and pollution evaluation in dry riverbeds affected by mine tailings.

The objective of this study was to evaluate the level of pollution, sources and potential risk of heavy metals (Zn, Cu, Mn, Cd, Cr, Ni, Fe and Pb) and arsenic (As) in four dry riverbeds affected by mine tailing, which drain into one of the biggest coastal lagoon of Europe (Mar Menor). El Beal, La Carrasquilla, Las Matildes and Ponce dry riverbeds sediments were sampled along its course (20, 18, 13, 19 samples were collected, respectively), and total/soluble metal(loid)s, water soluble ions, nitrogen, and organic/inorganic carbon contents were analyzed. Spatial distribution, principal component analysis (PCA), hierarchical cluster analysis (HCA), contamination factor (Cf), pollution load index (PLI) and potential ecological risk index (RI) were used to identify the possible sources of metal(loid)s and to assess the sediment pollution status. The results showed that the mean total concentrations of As, Cu, Cd, Mn, Zn and Pb exceeded the natural background levels of the study area, with the highest values located close to the mining areas. Correlation and cluster analysis identified that Cd and Zn were associated mainly with anthropogenic activities for all riverbeds, while Cr and Ni come from parent. PLI graded the four riverbeds as contaminated by heavy metals, while RI manifested that 100% of samples located in El Beal, La Carrasquilla and Las Matildes had a significantly high ecological risk. Therefore, this study suggests that mine wastes are the main source of metal(loids) contamination in the dry riverbeds, which results can be used to design actions and measures to reduce the environmental impact of metal(loid)s in the Mar Menor coastal lagoon.

Clinical phenotypes for risk stratification in small-for-gestational-age fetuses.

OBJECTIVE: To evaluate whether clinical phenotypes of small-for-gestational-age (SGA) fetuses can be identified and used for adverse perinatal outcome risk stratification to facilitate clinical decision-making. METHODS: This was a multicenter observational cohort study conducted in two tertiary care university hospitals. SGA fetuses were classified according to maternal, fetal and placental conditions using a two-step cluster algorithm, in which fetuses with more than one condition were assigned to the cluster associated with the highest mortality risk. Delivery and perinatal outcomes were compared using chi-square test among SGA clusters, and the associations between outcomes and each cluster were evaluated by calculating odds ratios (OR), adjusted for gestational age. RESULTS: The study included 17 631 consecutive singleton pregnancies, of which 1274 (7.2%) were defined as SGA at birth according to INTERGROWTH-21st standards. Nine SGA clinical phenotypes were identified using a predefined conceptual framework. All delivery and perinatal outcomes analyzed were significantly different among the nine phenotypes. The whole SGA cohort had a three-times higher risk of perinatal mortality compared with non-SGA fetuses (1.4% vs 0.4%; P < 0.001). SGA clinical phenotypes exhibited three patterns of perinatal mortality risk: the highest risk was associated with congenital anomaly (8.3%; OR, 17.17 (95% CI, 2.17-136.12)) and second- or third-trimester hemorrhage (8.3%; OR, 9.94 (95% CI, 1.23-80.02)) clusters; medium risk was associated with gestational diabetes (3.8%; OR, 9.59 (95% CI, 1.27-72.57)), preterm birth (3.2%; OR, 4.65 (95% CI, 0.62-35.01)) and intrauterine growth restriction (3.1%; OR, 5.93 (95% CI, 3.21-10.95)) clusters; and the lowest risk was associated with the remaining clusters. Perinatal mortality rate did not differ between SGA fetuses without other clinical conditions (54.1% of SGA fetuses) and appropriate-for-gestational-age fetuses (0.1% vs 0.4%; OR, 0.41 (95% CI, 0.06-2.94); P = 0.27). SGA combined with other obstetric pathologies increased significantly the risk of perinatal mortality, as demonstrated by the increased odds of perinatal death in SGA cases with gestational diabetes compared to non-SGA cases with the same condition (OR, 24.40 (95% CI, 1.31-453.91)). CONCLUSIONS: We identified nine SGA clinical phenotypes associated with different patterns of risk for adverse perinatal outcome. Our findings suggest that considering clinical characteristics in addition to ultrasound findings could improve risk stratification and decision-making for management of SGA fetuses. Future clinical trials investigating management of fetuses with SGA should take into account clinical information in addition to Doppler parameters and estimated fetal weight. © 2021 International Society of Ultrasound in Obstetrics and Gynecology.

Are the soils and vegetation of a forest close to tailings ponds affected by metals and arsenic?

This study aimed to determine the transfer of metals (Cd, Pb and Zn) and As to a Mediterranean forest close to five tailings ponds in Cartagena-La Union mining district (SE Spain). In addition, the effect of the rhizosphere of two native plant species, Olea europaea (OE) and Pistacia lentiscus (PL), on soil properties and chemical speciation of metal(oid)s was evaluated. Results showed there was no influence of the rhizosphere in the total concentration of metal(loid)s in soil, decreasing as Pb > Zn > As > Cd. Chemical partitioning revealed that only Cd and As can be considered hazardous, with a high percentage of these elements in the soil-labile fractions (20-40%). The accumulation in vegetal tissues was only high for Pb in PL roots, which makes it a suitable species for phytostabilization. Additionally, translocation factors showed transfer of Pb and Zn in OE, and Zn in PL to aerial parts, although no toxicity evidences for plants or animals were found. Finally, soil properties affected metal(loid)s accumulation in plants. The OE species was related to soil-labile metal(loid) fractions and pH, total N, organic carbon and silt content. The PL species were associated with immobilized metal(loid) fractions, sand content, electrical conductivity and total concentrations of As, Cd and Pb. Hence, mining activity has affected native adjacent soils, with accumulation of metals in plant species, although translocation was low, likely due to physiological strategies of the studied species to protect themselves against hazardous elements, and to the high soil pH, which limits metals' mobility.

MRP4-mediated cAMP efflux is essential for mouse spermatozoa capacitation.

Mammalian spermatozoa must undergo biochemical and structural changes to acquire the capacity for fertilization, in a process known as capacitation. Activation of PKA enzymes is essential for capacitation, and thus cAMP levels are tightly regulated during this process. Previously, we demonstrated that during capacitation, bovine spermatozoa extrude cAMP through multidrug resistance-associated protein 4 (MRP4, also known as ABCC4), which regulates intracellular levels of the nucleotide and provides cAMP to the extracellular space. Here, we report the presence of functional MRP4 in murine spermatozoa, since its pharmacological inhibition with MK571 decreased levels of extracellular cAMP. This also produced a sudden increase in PKA activity, with decreased tyrosine phosphorylation at the end of capacitation. Blockade of MRP4 inhibited induction of acrosome reaction, hyperactivation and in vitro fertilization. Moreover, MRP4 inhibition generated an increase in Ca2+ levels mediated by PKA, and depletion of Ca2+ salts from the medium prevented the loss of motility and phosphotyrosine inhibition produced by MK571. These results were supported using spermatozoa from CatSper Ca2+ channel knockout mice. Taken together, these results suggest that cAMP efflux via MRP4 plays an essential role in mouse sperm capacitation.This article has an associated First Person interview with the first author of the paper.

Sleep duration, physical activity, and caloric intake are related to weight status in Mexican American children: a longitudinal analysis.

BACKGROUND: Obesity is a serious issue, spanning all ages, and, in the U.S., disproportionately affects Latinos and African Americans. Understanding sleep, physical activity and dietary behaviors that may predict childhood obesity can help identify behavioral intervention targets. METHODS: Data were drawn from a U.S. cohort study of 323 Mexican American 8-10-year-old children and their mothers, who participated in a longitudinal study over a 2-year period. Measures were collected at baseline (BL; child mean age = 8.87, SD = 0.83), year 1 (FU1) and year 2 (FU2). Mothers reported on household income and acculturation at BL. Child height and weight were collected and BMI z-scores (BMIz) were calculated for weight status at BL, FU1, and FU2. Accelerometer-estimated sleep duration (hours) and moderate-to-vigorous physical activity (MVPA; minutes) were collected across 3 days at BL, FU1, and FU2. Two 24-h dietary recalls were performed at each time point; from these, average energy intake (EI, kcals/day) was estimated. Cross-lagged panel analysis was used to examine behavioral predictors on BMIz at each time point and across time. RESULTS: At BL and FU1, longer sleep duration (ß = - 0.22, p < 0.001; ß = - 0.17, p < 0.05, respectively) and greater MVPA (ß = - 0.13, p < 0.05; ß = - 0.20, p < 0.01, respectively) were concurrently related to lower BMIz. At FU2, longer sleep duration (ß = - 0.18, p < 0.01) was concurrently related to lower BMIz, whereas greater EI (ß = 0.16, p < 0.01) was related to higher BMIz. Longer sleep duration at BL predicted lower BMIz at FU1 (ß = - 0.05, p < 0.01). CONCLUSIONS: Longer sleep duration was concurrently related to lower weight status at each time point from ages 8-10 to 10-12. Higher MVPA was concurrently related to lower weight status in earlier childhood (ages 8-10 and 9-11) and higher EI was concurrently related to higher weight status toward the end of childhood (ages 10-12 years). Furthermore, longer sleep in earlier childhood was protective of children's lower weight status 1 year later. These findings suggest that sleep duration plays a consistent and protective role against childhood obesity; in addition, MVPA and healthy EI remain important independent factors for obtaining a healthy weight.

Enhanced photocatalytic performance and reusability of N-doped carbon dots/zinc oxide hybrid nanostructures.

We report the fabrication of nitrogen-doped carbon dots-zinc oxide hybrid (NCDs-ZnO) nanostructures utilizing simple chemical procedures. The role of NCDs in ZnO nanostructured matrix has been analyzed through XRD, SEM, FTIR and PL characterization techniques. The introduction of NCDs was found to modify not only their aspect ratio, observed by a reduction in the preferredc-axis growth compared to thea- andb-axis, but also induced an additional emission around 441 nm, which is typical of NCDs. The hybrid nanostructures were utilized as catalyst for methylene blue dye degradation showing a 95% degradation after 2 h of UV irradiation in comparison to the ∼70% degradation obtained by utilizing pristine ZnO, while the dye half-life (t1/2) was reduced by ∼65% by utilizing NCDs-ZnO hybrid nanostructures when compared to the pristine ZnO. The reusability of the fabricated hybrid structures was tested up to eight times with no significant loss in their photocatalytic performance (>90%). The stability of the hybrid structures was verified through Z-potential measurements prior and after reutilization. Excellent reusability and simple processing presented by NCDs-ZnO hybrid nanostructures makes them promising for industrial level photocatalyst for the waste water treatment.

Solving the migration-recombination equation from a genealogical point of view.

We consider the discrete-time migration-recombination equation, a deterministic, nonlinear dynamical system that describes the evolution of the genetic type distribution of a population evolving under migration and recombination in a law of large numbers setting. We relate this dynamics (forward in time) to a Markov chain, namely a labelled partitioning process, backward in time. This way, we obtain a stochastic representation of the solution of the migration-recombination equation. As a consequence, one obtains an explicit solution of the nonlinear dynamics, simply in terms of powers of the transition matrix of the Markov chain. The limiting and quasi-limiting behaviour of the Markov chain are investigated, which gives immediate access to the asymptotic behaviour of the dynamical system. We finally sketch the analogous situation in continuous time.

Comparative ecotoxicity of single and binary mixtures exposures of cadmium and nickel on growth and biomarkers of Lemna gibba.

The aim of the present study was to investigate the toxicity effects of cadmium-nickel (Cd-Ni) after single and mixtures exposures over the macrophyte Lemna gibba. Effects were assessed on growth, as frond number and fresh weight and biochemical parameters, such as total protein content and activity of antioxidant enzymes. Plants were exposed to single Cd and Ni in concentrations that ranged between 0.13-33 mg/L and 0.18 and 11.82 mg/L, respectively. For binary mixtures, individual metal IC50 values were used for selection of the evaluated concentrations. The experimental design consisted in three different ratios based on the concept of toxic units (TU), each ratio was evaluated by five different concentrations. Both single and mixture treatments were performed for 7 days following the conditions according to OECD (2006). Single and mixture exposures affected plant growth and the biomarkers of the antioxidant response. Growth parameters showed a differential sensitivity after individual metal exposures. Cd was more toxic for L. gibba plants when fresh weight was considered, but on the contrary, considering frond number, Ni was the most toxic metal. IC50-7d, based on growth rate calculated on frond number were 17.8 and 2.47 mg/L, and on fresh weight were 1.08 and 3.89 mg/L, for Cd and Ni respectively. LOEC values for Cd were obtained at 2.06 and 1.03 mg/L, for frond number and fresh weight, respectively; while for Ni, these values were 0.92 and 11.82 mg/L. The three evaluated ratios for binary mixtures resulted in a high toxicity considering the same response variables in single metal exposures. Ratio 1 (2/3 TU Cd-1/3 TU Ni) was the most toxic considering both frond number and fresh weight, showing percentage inhibition of growth rates of 96 and 90%, respectively for the highest concentration. A modification of the protein content was observed in single, but especially in the mixture treatments. The activity of catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APOX; EC 1.11.1.11) and guaiacol peroxidase (GPOX; EC 1.11.1.7) was also affected in single and mixtures assays. APOX and GPOX showed a higher increase of its activities respect the controls after mixture treatments than for single metal treatments. Such optimization of the antioxidant system could be one of the causes of the antagonistic toxicity observed in mixture exposures. Concentration addition (CA) reference model, based on frond number, in Cd-Ni mixtures was not a good predictor to evaluate toxicity from dissolved metal concentration since the results showed that toxicity was less than additive, with an average of ΣTU = 2.17. The observed antagonisms resulted to be stronger in mixtures with higher metal concentrations.

Smoking Behaviors of General Educational Development (GED) Recipients.

OBJECTIVE: Cigarette smoking is inversely associated with education, and despite this gradient effect, individuals with a General Educational Development (GED) diploma, obtained through a high school equivalency test, have the highest smoking prevalence. Considered the high school equivalency credential, it is unclear why individuals with a GED have a substantially higher smoking prevalence compared to high school graduates and dropouts. We conducted a qualitative study to understand life experiences, tobacco use patterns, and perceptions of tobacco among GED smokers and attitudes and behaviors around smoking cessation. METHODS: We recruited 40 GED smokers aged 18 to 35 years and conducted surveys and semi-structured individual interviews. Transcripts were independently coded, then coding differences were resolved and reviewed by a third team member. We independently determined themes within and between codes and met to determine final themes. RESULTS: GED recipients had many early life experiences and characteristics that made them highly vulnerable to tobacco dependence. With perceived high cognitive abilities, GED smokers were knowledgeable of many evidence-based smoking cessation strategies and were aware of health risks. Health risks and the financial burden of smoking were motivation to quit. The majority were uninterested or lacked confidence that nicotine replacement therapy, medications, counseling, or the quitline could help them quit. Nearly all were enthusiastic and confident that financial incentives had the potential to help them quit. CONCLUSIONS: Research is needed to determine whether financial incentives could improve smoking cessation outcomes in this unique population with an unequal burden of smoking.

Mimicking the temperature gradient between the sow's oviduct and uterus improves in vitro embryo culture output.

This work was designed to determine temperature conditions within the reproductive tract of the female pig and study their impact on ARTs. Temperatures were recorded using a laparo-endoscopic single-site surgery assisted approach and a miniaturized probe. Sows and gilts were used to address natural cycle and ovarian stimulation treatments, respectively. According to in vivo values, IVF was performed at three temperature conditions (37.0°C, 38.5°C and 39.5°C) and presumptive zygotes were cultured in these conditions for 20 h, while further embryo culture (EC) (21-168 h post-insemination) was maintained at 38.5°C. After 20 h, different fertility parameters were assessed. During EC, cleavage and blastocyst stages were evaluated. Sperm membrane fluidity at the experimental temperatures was studied by using differential scanning calorimetry and fluorescence recovery after photobleaching techniques. An increasing temperature gradient of 1.5°C was found between the oviduct and uterus of sows (P < 0.05) and when this gradient was transferred to pig in vitro culture, the number of poly-nuclear zygotes after IVF was reduced and the percentage of blastocysts was increased. Moreover, the temperature transition phase for the boar sperm membrane (37.0°C) coincided with the temperature registered in the sow oviduct, and sperm membranes were more fluid at 37.0°C compared with those of sperm incubated at higher temperatures (38.5°C and 39.5°C). These data suggest that there may be an impact of physiological temperature gradients on human embryo development.

Hemocyte cell types of the Cortes Geoduck, Panopea globosa (Dall 1898), from the Gulf of California, Mexico.

The geoduck Panopea globosa is an endemic and economic valuable species from the Mexican Northwest coast whose biology has been little studied. No information exists about their hemocytes to date, which is highly important to assess the welfare of wild and cultured organisms. In this study, hemocytes of adult P. globosa were characterized at the morphological, ultrastructural and functional level. The mean number of hemocytes in the hemolymph of P. globosa was 6 × 105 ± 2 × 105 cells mL-1. The cells were identified as granulocytes (Gr) and hyalinocytes (H). The former accounted for 28% of adhered cells in the hemolymph, measured 6-18 µm, showed numerous basophilic granules in the cytoplasm, with round and eccentric nuclei, and a nucleus:cytoplasm ratio of 0.44 ± 0.01. Hyalinocytes were the most abundant cells in the hemolymph of P. globosa (72% adhered cells) and were subdivided, according to their size, in small (Hs) 4-12 µm and large (HL) 6-18 µm. Hyalinocytes were eosinophilic round or ovoid cells with a central or eccentric nucleus, few or no granules in the cytoplasm and similar nucleus:cytoplasm ratio (Hs: 0.63 and HL: 061). Lysosomes and lipids were observed in Gr, while carbohydrates were the most abundant energy substrate in H. Both hemocytic cell types, mainly Gr, were capable to ingest particles and yield superoxide (P > 0.05). The present study shows for the first time the cell types, abundance and immune activities of hemocytes present in the hemolymph of P. globosa. This information provides a useful baseline to carry out further research on the cellular immune response of the clam to potential pathogens or changes in environmental factors.

Stem cell paracrine actions in tissue regeneration and potential therapeutic effect in human endometrium: a retrospective study.

OBJECTIVE: Determining genetic and paracrine mechanisms behind endometrial regeneration in Asherman's syndrome and endometrial atrophy (AS/EA) patients after autologous CD133+ bone marrow-derived stem cell (CD133+ BMDSC) transplantation. DESIGN: Retrospective study using human endometrial biopsies and mouse models. SETTING: Fundación-IVI, IIS-La Fe, Valencia, Spain. SAMPLES: Endometrial biopsies collected before and after CD133+ BMDSC therapy, from eight women with AS/EA (NCT02144987) from the uterus of five mice with only left horns receiving CD133+ BMDSC therapy. METHODS: In human samples, haematoxylin and eosin (H&E) staining, RNA arrays, PCR validation, and neutrophil elastase (NE) immunohistochemistry (IHQ). In mouse samples, PCR validation and protein immunoarrays. MAIN OUTCOME MEASURES: H&E microscopic evaluation, RNA expression levels, PCR, and growth/angiogenic factors quantification, NE IHQ signal. RESULTS: Treatment improved endometrial morphology and thickness for all patients. In human samples, Jun, Serpine1, and Il4 were up-regulated whereas Ccnd1 and Cxcl8 were down-regulated after treatment. The significant decrease of NE signal corroborated Cxcl8 expression. Animal model analysis confirmed human results and revealed a higher expression of pro-angiogenic cytokines (IL18, HGF, MCP-1, MIP2) in treated uterine horns. CONCLUSIONS: CD133+ BMDSC seems to activate several factors through a paracrine mechanism to help tissue regeneration, modifying endometrial behaviour through an immunomodulatory milieu that precedes proliferation and angiogenic processes. Insight into these processes could bring us one step closer to a non-invasive treatment for AS/EA patients. TWEETABLE ABSTRACT: CD133+ BMDSC therapy regenerates endometrium, modifying the immunological milieu that precedes proliferation and angiogenesis.

Clinical impact of Doppler reference charts on management of small-for-gestational-age fetuses: need for standardization.

OBJECTIVE: To assess clinical variability in the management of small-for-gestational-age (SGA) fetuses according to different published Doppler reference charts for umbilical artery (UA) and fetal middle cerebral artery (MCA) Doppler indices and cerebroplacental ratio (CPR). METHODS: We performed a systematic search of MEDLINE, EMBASE, CINAHL and the Web of Science databases from 1954 to 2018 for studies with the sole aim of creating fetal Doppler reference values for UA, MCA and CPR. The top cited articles for each Doppler parameter were included. Variability in Doppler values at the following clinically relevant cut-offs was assessed: UA-pulsatility index (PI) > 95th percentile; MCA-PI < 5th percentile; and CPR < 5th percentile. Variability was calculated for each week of gestation and expressed as the percentage difference between the highest and lowest Doppler value at the clinically relevant cut-offs. Simulation analysis was performed in a cohort of SGA fetuses (n = 617) to evaluate the impact of this variability on clinical management. RESULTS: From a total of 40 studies that met the inclusion criteria, 19 were analyzed (13 for UA-PI, 10 for MCA-PI and five for CPR). Wide discrepancies in reported Doppler reference values at clinically relevant cut-offs were found. MCA-PI showed the greatest variability, with differences of up to 51% in the 5th percentile value at term. Variability in the 95th percentile of UA-PI and the 5th percentile of CPR at each gestational week ranged from 21% to 41% and 15% to 33%, respectively. As expected, on simulation analysis, these differences in Doppler cut-off values were associated with significant variation in the clinical management of SGA fetuses, despite using the same protocol. CONCLUSIONS: The choice of Doppler reference chart can result in significant variation in the clinical management of SGA fetuses, which may lead to suboptimal outcomes and inaccurate research conclusions. Therefore, an attempt to standardize fetal Doppler reference ranges is needed. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd.

Photodynamic therapy (PDT) in non-surgical treatment of periodontitis.

Periodontitis represents a major problem for patients, since it is not possible to eliminate the bacteria that are responsible for this pathology with a pharmacological treatment. The present study included forty-four patients with periodontitis, who had undergone disinfection via photodynamic therapy (PDT) using a laser source having a 635 nm wavelength associated with a photoactivable substance (methylene blue). Clinical assessment of plaque index (PI), bleeding on probing (BOP), probing depth (PD), calculus index (CI), gingival recession (REC) and clinical attachment level (CAL) were recorded at base line, 1 month (4 weeks) after treatment and again 3 months (12 weeks) after treatment, while site radiography (RX) and microbiological test (MT) were recorded at base line and 3 months (12 weeks) after treatment. The outcomes show a good efficacy of the PDT in the elimination of the periodontal pathogenic microflora and in the improvement of the clinical parameters considered: from the base line to the final check after 12 weeks it has been observed a reduction in REC of about 16.9%, a reduction of CAL of about 17.85%, a reduction of the BoP of about 93.3%, a reduction of the PD of about 17%, a reduction of the CI of about 66.3%, a reduction of PI of about 44%, and microbiologically a reduction of the total amount of bacteria with proven parodontopathic properties (red complex bacteria) of about 58.74%. Within the limits of the present study, PDT can be reasonably considered as a good carrier that leads to significant improvements in the parameters (clinical and microbiological) considered.

Virulence properties and antimicrobial resistance of Pseudomonas aeruginosa isolated from cave waters at Roraima Tepui, Guayana Highlands.

Pseudomonas aeruginosa is a prominent member of emerging waterborne pathogens. The environmental reservoirs of multi-resistant phenotypes and other virulence factors in this bacterium are poorly understood. Our study aimed to determine the virulence properties of P. aeruginosa isolated from Roraima Sur Cave (RSC) waters at Guayana Highlands. Based on the best identification at species level by biochemical tests, 16S rRNA sequencing and phylogenetic inferences, one RSC isolate named LG11 was characterized for virulence properties in comparison with P. aeruginosa reference strains. PCR amplification of alginate, elastase, exoenzyme S, exotoxin A, neuraminidase and Quorum-Sensing genes showed a high virulence potential in LG11. This isolate demonstrated multi-resistance to ceftriaxone, tigecycline and imipenem. Pyocyanin production was greater in LG11 (0·478 µg ml-1 ) than the strain ATCC 10145 (0·316 µg ml-1 ), but the highest pigment concentration (2·140 µg ml-1 ) was displayed by the clinical strain CVCM 937 (P = 0·000175). Pronounced biomass production on granite and glass (P < 0·05) and well-developed biofilms indicated the ability of P. aeruginosa from RSC to colonize surfaces found in human and healthcare environments. These data suggest that waters from pristine ecosystems such as RSC could be reservoirs of this opportunistic bacterium carrying important virulence properties with potential epidemiological implications. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows for the first time the occurrence of virulence genes and multi-resistance to antimicrobials in Pseudomonas aeruginosa isolated from cave waters at Guayana Highlands. These findings, together with the biofilm formation on surfaces found in human and healthcare settings, suggest public health risks and the potential of these virulence properties to be transferred from or to native populations in waters. Our results provide important insights to the current knowledge of P. aeruginosa in the environment, setting the basis for future studies driven to assess reservoirs of multi-resistant bacteria and virulence features unknown in pristine ecosystems.