Having children in cross-border contexts: late-family formation among homoparental families in Spain.

Considered until recently unfit to rear children, non-heterosexual people have been excluded from forming families in most countries. Many, worldwide, demand access to family formation, claiming the same aptitudes as heterosexual people for raising children. However, when non-heterosexual singles and couples want to become parents in Spain, they must consider transnational contexts, resorting to inter-country adoption or surrogacy abroad, processes that contribute to delay their family formation. They must consider not only Spanish sociocultural conditions, but other countries' legal restrictions regarding parents' gender, social status, and sexual identity. These families experience great difficulty in gaining access to reproductive health services. Based on multi-site ethnographic fieldwork, this text addresses how, despite legislative changes allowing homoparental family formation in Spain, these parents must overcome complex bureaucratic processes when they decide to have children, while facing homophobic attitudes and policies in their quests to become parents.

Multivariate data analysis of process parameters affecting the growth and productivity of stable Chinese hamster ovary cell pools expressing SARS-CoV-2 spike protein as vaccine antigen in early process development.

The recent COVID-19 pandemic revealed an urgent need to develop robust cell culture platforms which can react rapidly to respond to this kind of global health issue. Chinese hamster ovary (CHO) stable pools can be a vital alternative to quickly provide gram amounts of recombinant proteins required for early-phase clinical assays. In this study, we analyze early process development data of recombinant trimeric spike protein Cumate-inducible manufacturing platform utilizing CHO stable pool as a preferred production host across three different stirred-tank bioreactor scales (0.75, 1, and 10 L). The impact of cell passage number as an indicator of cell age, methionine sulfoximine (MSX) concentration as a selection pressure, and cell seeding density was investigated using stable pools expressing three variants of concern. Multivariate data analysis with principal component analysis and batch-wise unfolding technique was applied to evaluate the effect of critical process parameters on production variability and a random forest (RF) model was developed to forecast protein production. In order to further improve process understanding, the RF model was analyzed with Shapley value dependency plots so as to determine what ranges of variables were most associated with increased protein production. Increasing longevity, controlling lactate build-up, and altering pH deadband are considered promising approaches to improve overall culture outcomes. The results also demonstrated that these pools are in general stable expressing similar level of spike proteins up to cell passage 11 (~31 cell generations). This enables to expand enough cells required to seed large volume of 200-2000 L bioreactor.

Low-Density Lipoproteins Increase Proliferation, Invasion, and Chemoresistance via an Exosome Autocrine Mechanism in MDA-MB-231 Chemoresistant Cells.

Dyslipidemias involving high concentrations of low-density lipoproteins (LDLs) increase the risk of developing triple-negative breast cancer (TNBC), wherein cholesterol metabolism and protein translation initiation mechanisms have been linked with chemoresistance. Doxorubicin (Dox) treatment, a member of the anthracycline family, represents a typical therapeutic strategy; however, chemoresistance remains a significant challenge. Exosomes (Exs) secreted by tumoral cells have been implicated in cell communication pathways and chemoresistance mechanisms; the content of exosomes is an outcome of cellular cholesterol metabolism. We previously induced Dox resistance in TNBC cell models, characterizing a variant denominated as variant B cells. Our results suggest that LDL internalization in parental and chemoresistant variant B cells is associated with increased cell proliferation, migration, invasion, and spheroid growth. We identified the role of eIF4F translation initiation factor and the down-regulation of tumor suppressor gene PDCD4, an inhibitor of eIF4A, in chemoresistant variant B cells. In addition, the exomes secreted by variant B cells were characterized by the protein content, electronic microscopy, and cell internalization assays. Critically, exosomes purified from LDL-treated variant B cell promoted cell proliferation, migration, and an increment in lactate concentration. Our results suggest that an autocrine phenomenon induced by exosomes in chemoresistant cells may induce modifications on signaling mechanisms of the p53/Mdm2 axis and activation of p70 ribosomal protein kinase S6. Moreover, the specific down-regulated profile of chaperones Hsp90 and Hsp70 secretion inside the exosomes of the chemoresistant variant could be associated with this phenomenon. Therefore, autocrine activation mediated by exosomes and the effect of LDL internalization may influence changes in exosome chaperone content and modulate proliferative signaling pathways, increasing the aggressiveness of MDA-MB-231 chemoresistant cells.

Brucella abortus triggers the differential expression of immunomodulatory lncRNAs in infected murine macrophages.

Introduction: The lncRNAs (long non-coding RNAs) are the most diverse group of non-coding RNAs and are involved in most biological processes including the immune response. While some of them have been recognized for their influence on the regulation of inflammatory activity, little is known in the context of infection by Brucella abortus, a pathogen that presents significant challenges due to its ability to manipulate and evade the host immune system. This study focuses on characterize the expression profile of LincRNA-cox2, Lethe, lincRNA-EPS, Malat1 and Gas5 during infection of macrophages by B. abortus. Methods: Using public raw RNA-seq datasets we constructed for a lncRNA expression profile in macrophages Brucella-infected. In addition, from public RNA-seq raw datasets of RAW264.7 cells infected with B. abortus we constructed a transcriptomic profile of lncRNAs in order to know the expression of the five immunomodulating lncRNAs studied here at 8 and 24 h post-infection. Finally, we performed in vitro infection assays in RAW264.7 cells and peritoneal macrophages to detect by qPCR changes in the expression of these lncRNAs at first 12 hours post infection, a key stage in the infection cycle where Brucella modulates the immune response to survive. Results: Our results demonstrate that infection of macrophages with Brucella abortus, induces significant changes in the expression of LincRNA-Cox2, Lethe, LincRNA-EPS, Gas5, and Malat1. Discussion: The change in the expression profile of these immunomodulatory lncRNAs in response to infection, suggest a potential involvement in the immune evasion strategy employed by Brucella to facilitate its intracellular survival.

Pseudoachalasia as a paraneoplastic event in a patient with Hodgkin's disease.

Pseudoachalasia or secondary achalasia (5% of achalasias that are deemed primary achalasias) is an esophageal motor disorder with manometric criteria for achalasia, but it appears in the context of an underlying pathology that can be attributed to its origin. Usually appears in >60 years with rapid evolution of symptoms (<1 year). The main cause of pseudoachalasia is neoformative etiology, but there are others. Our patient started with rapid progression dysphagia and was diagnosed with type II achalasia within a Hodgkin's lymphoma. In the radiological-metabolic studies, disease involvement was ruled out as an extrinsic compression of the esophagogastric junction as well as signs of its activity at this level. Chemotherapy has not been shown to play a role in the development of this pathology. On the other hand, radiotherapy has been associated with an esophageal motor disorder, but, in our case, it was after its onset. Therefore, we propose that the mechanism of pseudoachalasia in our case is a paraneoplastic event. This hypothesis is related to other similar cases reported, and it reflects the importance of continuing to investigate this clinical condition that is indistinguishable by manometry from primary achalasia. In addition, it usually presents differential clinical characteristics whose early recognition has implications for the diagnostic, therapeutic, and prognostic management of the patient.

Pseudoachalasia as a paraneoplastic event in a patient with Hodgkin’s disease

Pseudoachalasia or secondary achalasia (5% of achalasias that are deemed primary achalasias) is an esophageal motor disorder with manometric criteria for achalasia, but it appears in the context of an underlying pathology that can be attributed to its origin. Usually appears in >60 years with rapid evolution of symptoms (<1 year). The main cause of pseudoachalasia is neoformative etiology, but there are others. Our patient started with rapid progression dysphagia and was diagnosed with type II achalasia within a Hodgkin's lymphoma. In the radiological-metabolic studies, disease involvement was ruled out as an extrinsic compression of the esophagogastric junction as well as signs of its activity at this level. Chemotherapy has not been shown to play a role in the development of this pathology. On the other hand, radiotherapy has been associated with an esophageal motor disorder, but, in our case, it was after its onset. Therefore, we propose that the mechanism of pseudoachalasia in our case is a paraneoplastic event. This hypothesis is related to other similar cases reported, and it reflects the importance of continuing to investigate this clinical condition that is indistinguishable by manometry from primary achalasia. In addition, it usually presents differential clinical characteristics whose early recognition has implications for the diagnostic, therapeutic, and prognostic management of the patient. (AU)

Simultaneous Bilateral Video-Endoscopic Inguinal Lymphadenectomy for Penile Carcinoma: Surgical Setting, Feasibility, Safety, and Preliminary Oncological Outcomes.

INTRODUCTION: Inguinal lymph node dissection (ILND) plays an important role for both staging and treatment purposes in patients diagnosed with penile carcinoma (PeCa). Video-endoscopic inguinal lymphadenectomy (VEIL) has been introduced to reduce complications, and in those patients elected for bilateral ILND, a simultaneous bilateral VEIL (sB-VEIL) has also been proposed. This study aimed to investigate the feasibility, safety, and preliminary oncological outcomes of sB-VEIL compared to consecutive bilateral VEIL (cB-VEIL). MATERIAL AND METHODS: Clinical N0-2 patients diagnosed with PeCa and treated with cB-VEIL and sB-VEIL between 2015 and 2023 at our institution were included. Modified ILND was performed in cN0 patients, while cN+ patients underwent a radical approach. Intra- and postoperative complications, operative time, time of drainage maintenance, length of hospital stay and readmission within 90 days, as well as lymph node yield, were compared between the two groups. RESULTS: Overall, 30 patients were submitted to B-VEIL. Of these, 20 and 10 patients underwent cB-VEIL and sB-VEIL, respectively. Overall, 16 (80%) and 7 (70%) patients were submitted to radical ILND due to cN1-2 disease in the cB-VEIL and sB-VEIL groups, respectively. No statistically significant difference emerged in terms of median nodal yield (13.5 vs. 14, p = 0.7) and median positive LNs (p = 0.9). sD-VEIL was associated with a shorter operative time (170 vs. 240 min, p < 0.01). No statistically significant difference emerged in terms of intraoperative estimated blood loss, length of hospital stay, time to drainage tube removal, major complications, and hospital readmission in the cB-VEIL and sB-VEIL groups, respectively (all p > 0.05). CONCLUSIONS: Simultaneous bilateral VEIL is a feasible and safe technique in patients with PeCA, showing similar oncological results and shorter operative time compared to a consecutive bilateral approach. Patients with higher preoperative comorbidity burden or anesthesiological risk are those who may benefit the most from this technique.

Seasonal heat stress compromises testicular thermoregulation and semen quality of Dorper rams raised in a desert climate.

Dorper rams are widely distributed throughout the world under different climatic conditions, however, little is known about their reproductive performance in desert regions. Ten Dorper rams were individually housed and exposed to thermoneutrality for 35 d in spring (23.6 ± 5.6 °C, mean ± SD) and outdoor heat stress (HS) for 35 d in summer (33.6 ± 2.0 °C) to evaluate the effect of seasonal HS on physiological responses, testicular biometry, and seminal quality under desert climatic conditions. Rectal temperature, respiration rate and coat surface temperatures in different body regions were measured every 7 d (0600, 1200, and 1800 h); also, testicular biometry was registered at 0600 h. Semen was collected via an artificial vagina 3 d after physiological variables were measured and seminal traits were evaluated. Rectal temperature, respiration rate and coat surface temperatures were higher (P < 0.01) at each hour of measurement in summer compared to spring. Overall, scrotal length and circumference, as well as testicular volume were higher (P < 0.01) in summer than in spring. Compared to spring conditions, summer HS caused lower (P ≤ 0.05) sperm concentration and viability combined with a higher percentage of sperm abnormalities without affecting ejaculate volume. Both mass and sperm motility were similar between seasons in the first two sampling weeks, and then decreased (P ≤ 0.03) due to summer HS. In conclusion, Dorper rams developed testicle hyperthermia and, consequently, showed poor semen quality due to the high environmental temperatures prevailing in desert regions.

Recurrent hemobilia secondary to extrahepatic biliary tract cholangiocarcinoma. A diagnostic challenge.

The etiology of hemobilia has mainly iatrogenic (>50%), followed by traumatic causes. Others are biliopathy due to portal high pressure, or neoplastic or infective biliopathy. In the case of non-clear hemobilia, direct-vision-cholangioscopy can change the management in >34% of cases. Our patient had episodes of obstructive hemobilia with secondary cholangitis without objectifying underlying pathology. When she was referred to our center, SpyGlass®-cholangioscopy identified the suspicious lesion compatible with early-stage cholangiocarcinoma despite the diagnostic delay. In conclusion, it is important to keep in mind the neoformative etiology as a potential cause of hemobilia of unclear origin, in which case, cholangioscopy (SpyGlass®) can contribute to the recognition of the signs of malignancy of the lesion and, therefore, to the diagnosis.

A novel mechanistic modelling approach for microbial selection dynamics: Towards improved design and control of raceway reactors for purple bacteria.

Purple phototrophic bacteria (PPB) show an underexplored potential for resource recovery from wastewater. Raceway reactors offer a more affordable full-scale solution on wastewater and enable useful additional aerobic processes. Current mathematical models of PPB systems provide useful mechanistic insights, but do not represent the full metabolic versatility of PPB and thus require further advancement to simulate the process for technology development and control. In this study, a new modelling approach for PPB that integrates the photoheterotrophic, and both anaerobic and aerobic chemoheterotrophic metabolic pathways through an empirical parallel metabolic growth constant was proposed. It aimed the modelling of microbial selection dynamics in competition with aerobic and anaerobic microbial community under different operational scenarios. A sensitivity analysis was carried out to identify the most influential parameters within the model and calibrate them based on experimental data. Process perturbation scenarios were simulated, which showed a good performance of the model.

Unraveling the Role of the Zinc-Dependent Metalloproteinase/HTH-Xre Toxin/Antitoxin (TA) System of Brucella abortus in the Oxidative Stress Response: Insights into the Stress Response and Virulence.

Toxin/antitoxin (TA) systems have been scarcely studied in Brucella abortus, the causative agent of brucellosis, which is one of the most prevalent zoonotic diseases worldwide. In this study, the roles of a putative type II TA system composed by a Zinc-dependent metalloproteinase (ZnMP) and a transcriptional regulator HTH-Xre were evaluated. The deletion of the open reading frame (ORF) BAB1_0270, coding for ZnMP, used to produce a mutant strain, allowed us to evaluate the survival and gene expression of B. abortus 2308 under oxidative conditions. Our results showed that the B. abortus mutant strain exhibited a significantly reduced capacity to survive under hydrogen peroxide-induced oxidative stress. Furthermore, this mutant strain showed a decreased expression of genes coding for catalase (katE), alkyl hydroperoxide reductase (ahpC) and transcriptional regulators (oxyR and oxyR-like), as well as genes involved in the general stress response, phyR and rpoE1, when compared to the wild-type strain. These findings suggest that this type II ZnMP/HTH-Xre TA system is required by B. abortus to resist oxidative stress. Additionally, previous evidence has demonstrated that this ZnMP also participates in the acidic stress resistance and virulence of B. abortus 2308. Therefore, we propose a hypothetical regulatory function for this ZnMP/HTH-Xre TA system, providing insight into the stress response and its potential roles in the pathogenesis of B. abortus.

Achieving Discharge Limits in Single-Stage Domestic Wastewater Treatment by Combining Urban Waste Sources and Phototrophic Mixed Cultures.

This work shows the potential of a new way of co-treatment of domestic wastewater (DWW) and a liquid stream coming from the thermal hydrolysis of the organic fraction of municipal solid waste (OFMSW) mediated by a mixed culture of purple phototrophic bacteria (PPB) capable of assimilating carbon and nutrients from the medium. The biological system is an open single-step process operated under microaerophilic conditions at an oxidative reduction potential (ORP) < 0 mV with a photoperiod of 12/24 h and fed during the light stage only so the results can be extrapolated to outdoor open pond operations by monitoring the ORP. The effluent mostly complies with the discharge values of the Spanish legislation in COD and p-values (<125 mg/L; <2 mg/L), respectively, and punctually on values in N (<15 mg/L). Applying an HRT of 3 d and a ratio of 100:7 (COD:N), the presence of PPB in the mixed culture surpassed 50% of 16S rRNA gene copies, removing 78% of COD, 53% of N, and 66% of P. Furthermore, by increasing the HRT to 5 d, removal efficiencies of 83% of COD, 65% of N, and 91% of P were achieved. In addition, the reactors were further operated in a membrane bioreactor, thus separating the HRT from the SRT to increase the specific loading rate. Very satisfactory removal efficiencies were achieved by applying an HRT and SRT of 2.3 and 3 d, respectively: 84% of COD, 49% of N, and 93% of P despite the low presence of PPB due to more oxidative conditions, which step-by-step re-colonized the mixed culture until reaching >20% of 16S rRNA gene copies after 49 d of operation. These results open the door to scaling up the process in open photobioreactors capable of treating urban wastewater and municipal solid waste in a single stage and under microaerophilic conditions by controlling the ORP of the system.

A Triazaspirane Derivative Inhibits Migration and Invasion in PC3 Prostate Cancer Cells.

Cancer is a serious health problem due to the complexity of establishing an effective treatment. The purpose of this work was to evaluate the activity of a triazaspirane as a migration and invasion inhibitor in PC3 prostatic tumor cells through a possible negative regulation of the FAK/Src signal transduction pathway and decreased secretion of metalloproteinases 2 and 9. Molecular docking analysis was performed using Moe 2008.10 software. Migration (wound-healing assay) and invasion (Boyden chamber assay) assays were performed. In addition, the Western blot technique was used to quantify protein expression, and the zymography technique was used to observe the secretion of metalloproteinases. Molecular docking showed interactions in regions of interest of the FAK and Src proteins. Moreover, the biological activity assays demonstrated an inhibitory effect on cell migration and invasion, an important suppression of metalloproteinase secretion, and a decrease in the expression of p-FAK and p-Src proteins in treated PC3 cells. Triazaspirane-type molecules have important inhibitory effects on the mechanisms associated with metastasis in PC3 tumor cells.

Mesalazine-induced esophageal ulcers. a rare adverse effect.

31-year-old woman. Diagnosis of ulcerative proctitis in February/2022. Calprotectin 1832 µg/g. Colonoscopy: erythematous, friable and erosive mucosa up to 10 cm from the anal margin. Pathology: compatible with ulcerative colitis with moderate activity. Start of oral mesalazine (3 gr/24 h granules) and topical (1 gr/24 h suppository). After three months, she achieved clinical remission. Calprotectin 57 µg/g. Two months later, she consulted for solid dysphagia, loss of 10 kg, and low-grade fever for a month. Fifteen days before, she went to an emergency room where Prednisone 50 mg/24 h was started. On the day of the assessment, she was receiving 30 mg with no improvement. The next day, gastroscopy showed 6-12 mm esophageal ulcers with non-confluent shallow geographic borders, biopsies were taken. Viral serologies and HLA B51 were requested. Given the severity of the symptoms, empirical treatment was started with Valaciclovir 1 g/12 h. Serologies: IgG for Ebstein Barr virus, cytomegalovirus and herpes virus with negative IgM. Cytomegalovirus viral load: <30 IU/ml. Pathology: acute extensively ulcerated esophagitis, inflammatory infiltrate and some eosinophils with negative histochemical staining for fungi, cytomegalovirus and herpes virus I and II. HLA B51 was negative. Valaciclovir and mesalazine are discontinued after seven days given the known relationship of the latter with low-grade fever and, exceptionally, with esophageal pathology. Three days later, the patient reported clear improvement in dysphagia from the day the mesalazine was discontinued. After eight months, she was still asymptomatic. Upon resolution of the symptoms, control gastroscopy was not performed, and mesalazine has not been reintroduced due to its probable causal association. Mesalazine has an excellent safety profile. Adverse effects include fever, headache, diarrhea and.

Potential Inhibitors of The OTUB1 Catalytic Site to Develop an Anti-Cancer Drug Using In-Silico Approaches.

Background: : Cancer continues worldwide. It has been reported that OTUB1, a cysteine protease, plays a critical role in a variety of tumors and is strongly related to tumor proliferation, migration, and clinical prognosis by its functions on deubiquitination. Drug advances continue against new therapeutic targets. In this study we used OTUB1 to develop a specific pharmacological treatment to regulate deubiquitination by OTUB1. The aim of this research is to regulate OTUB1 functions. Methods: By molecular docking in a specific potential OTUB1 interaction site between Asp88, Cys91, and His26 amino acids, using a chemical library of over 500,000 compounds, we selected potential inhibitors of the OTUB1 catalytic site. Results: Ten compounds (OT1 - OT10) were selected by molecular docking to develop a new anti-cancer drug to decrease OTUB1 functions in cancer processes. Conclusion: OT1 - OT10 compounds could be interacting in the potential site between Asp88, Cys91, and His265 amino acids in OTUB1. This site is necessary for the deubiquitinating function of OTUB1. Therefore, this study shows another way to attack cancer.

To drain or not to drain in uro-oncological robotic surgery? A systematic review and meta-analysis.

INTRODUCTION: The aim of this study was to compare the perioperative outcomes of routine drainage insertion vs. no drainage in patients undergoing robot-assisted radical prostatectomy (RARP), robot-assisted partial nephrectomy (RAPN), and robot-assisted radical cystectomy (RARC). EVIDENCE ACQUISITION: A literature search was conducted through April 2022 using PubMed/Medline, Embase, and Web of Science databases. Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines were followed to identify eligible studies. EVIDENCE SYNTHESIS: Eleven studies comprising 8447 RARPs and 1890 RAPNs met our inclusion criteria. Our search strategy did not identify any studies within the RARC framework. In RARP, patients without postoperative drainage had lower rate of postoperative ileus (OR 0.53, 95% CI: 0.38 to 0.74; P<0.001) and similar low-grade (Clavien 1-2, P=0.41) and high-grade (Clavien ≥3; P=0.85) complications, urinary leakage (P=0.07), pelvic hematoma (P=0.35), symptomatic lymphocele (P=0.13), fever (P=0.25), incisional hernia (P=0.31), reintervention (P=0.57), length of hospital stay (P=0.22), and readmission (P=0.74) compared with routinely drained patients. In RAPN, patients without postoperative drainage had shorter length of hospital stay (mean difference: -0.84 days, 95% CI: -1.06 to -0.63; P<0.001) and similar low-grade (P=0.94) and high-grade (P=0.31) complications, urinary leakage (P=0.49), hemorrhage (P=0.39), reintervention (P=0.69), and readmission (P=0.20) compared with routinely drained patients. CONCLUSIONS: In our study, patients without drainage had similar perioperative course to patients with prophylactic drain insertion after RARP and RAPN. Omission of drain insertion was associated with a lower rate of postoperative ileus for RARP and a shorter hospital stay for RAPN. In the era of robotic surgery, routine drain placement is no longer indicated in unselected patients.

Histiocytic sarcoma of the esophagus.

A 20-year-old male with no medical history of interest who goes to the emergency room because of retrosternal pain, odynophagia, dysphagia, and fever. On physical examination: 37.7ºC axillary temperature, bad general condition, and central chest pain on palpation. In the blood test: 16,200x10^6/L white blood cells, 12,800x10^6/L neutrophils, and 11.66mg/dL C reactive protein, with the rest of the complete blood count, coagulation, and biochemistry within normal values.

Ferulic acid supplementation for 40 days in hair ewe lambs experiencing seasonal heat stress: short-term effects on physiological responses, growth, metabolism, and hematological profile.

Free ferulic acid (FA) is a natural compound with antioxidant properties which mitigates the negative effects of cold stress in sheep; however, its impact on thermoregulatory responses in heat-stressed sheep has not been defined. The objective was to evaluate the effects of FA supplementation on physiological responses, serum analyte concentrations, and the hematological profile of heat-stressed hair ewe lambs. Twenty-two Dorper × Katahdin ewe lambs (initial body weight = 23.5 ± 2.8 kg and age = 4 months) were housed in individual pens for 40 days and assigned under a randomized complete block design to the following treatments (n = 11): basal diet with 0 (control) or 250 mg of FA/kg of feed. The FA × sampling day interaction only affected serum concentration of some metabolic hormones; particularly on day 20 of the trial, FA increased (P < 0.01) insulins and the insulin to glucose ratio while decreased (P = 0.05) thyroxine. Overall, supplemental FA did not affect rectal temperature, respiratory rate, most body surface temperatures, feedlot performance, and serum concentrations of metabolites, electrolytes, triiodothyronine, and cortisol. In addition, FA only tended to decrease (P ≥ 0.09) erythrocyte count and plaquetocrit and to increase (P = 0.08) mean corpuscular volume. In conclusion, FA supplementation did not improve the growth nor thermoregulatory capacity of heat-stressed hair ewe lambs. Still, it partially modulated the metabolism to reinforce some energetic adaptive mechanisms when the ambient temperature was ≥ 35 °C.