Impact of the COVID-19 lockdown on the O'Sullivan test and gestational diabetes mellitus diagnosis in pregnant Spanish women.

OBJECTIVE: To analyze the impact of COVID-19 lockdown on serum glucose levels of pregnant women. DESIGN: A retrospective analysis of O'Sullivan test in pregnant women who underwent COVID-19 lockdown compared to controls. SITE: Poniente Primary Health Care center in Córdoba (Spain). PARTICIPANTS: 235 pregnant women from 23+0 to 25+0 weeks of gestation without diabetes mellitus. INTERVENTIONS: Gestational diabetes mellitus screening with O'Sullivan test and 3-h oral glucose tolerance test. MAIN MEASUREMENTS: Pregnant women who underwent gestational diabetes mellitus screening with O'Sullivan test before (control group) and during COVID-19 Lockdown (Lockdown group) in Córdoba (Spain) were investigated. Lockdown group was divided in early and late lockdown. An additional, control group from data of the same months of the Lockdown in the previous year were recorded to discarded seasonally (adjusted seasonally control) this group was also divided in early and late seasonally adjusted. A logistic regression model for O'Sullivan test has been performed to analyze potential cofounders. Kolgomorov-Smirnov and Kruskal-Wallis test comparing pregnant women who underwent COVID-19 lockdown with the two types of controls. RESULTS: Statistically significant differences were found in serum glucose after O'Sullivan test between lockdown group and control group (123.51±26.02mg/dL and 112.86±31.28mg/dL; p=0.017). When early lockdown group and control group were compared no differences were found (119.64±26.18mg/dL vs. 112.86±31.28mg/dL; p>0.05) whereas differences were observed in late lockdown group and control group (127.22±25.59mg/dL vs. 112.86±31.28mg/dL; p=0.009). Statistical trends were also found between lockdown group and seasonally adjusted group and between lockdown and late seasonally adjusted group (p=0.089). A higher proportion of positive O'Suvillan pregnant women who were subsequently diagnosed with GDM were found in lockdown group compared to the seasonally adjusted control group (60% vs. 26.06% respectively; p<0.05). CONCLUSIONS: The COVID-19 lockdown was associated with an increase in serum glucose levels after the O'Sullivan test as well as a higher GDM diagnosis risk in pregnant women. The findings of our study emphasize the essential requirement for comprehensive maternal services and the accessibility to community's health assets during future lockdown scenarios to pregnant women.

Parental presence during invasive pediatric procedures: what does it depend on? / A presença dos pais durante procedimentos pediátricos invasivos: depende de quê? / Presencia de los padres durante procedimientos pediátricos invasivos: ¿De qué depende?

Abstract Objective: family-centered care during invasive procedures has been endorsed by many professional health care organizations. The aim of this study was to evaluate the health professionals' attitudes towards parental presence during their child's invasive procedure. Method: pediatric healthcare providers (divided in professional categories and range of ages) from one of the Spain's largest hospitals were asked to complete a questionnaire and write free-text comments. Results: 227 responded the survey. Most (72%) participants, in their answers, reported that parents are sometimes present during interventions, although there were differences between professional categories in this respect. The procedures in which the parents were present were those considered "less invasive" (96% of cases), while only 4% were present in those considered "more invasive". The older the professional, the less necessary parental presence was considered. Conclusion: the attitudes towards parental presence during pediatric invasive procedure are influenced by the professional category, the age of the healthcare provider and the invasiveness of the procedure.

Resumo Objetivo: o atendimento centrado na família durante procedimentos invasivos tem sido endossado por muitas organizações profissionais de saúde. O objetivo deste estudo foi avaliar as atitudes dos profissionais de saúde em relação à presença dos pais durante o procedimento invasivo realizado em seus filhos. Método: os prestadores de serviços de saúde pediátricos (divididos em categorias profissionais e faixa etária) de um dos maiores hospitais da Espanha foram solicitados a preencher um questionário e escrever comentários de texto livre. Resultados: a pesquisa foi respondida por 227 pessoas. A maioria (72%) dos participantes, em suas respostas, relatou que os pais algumas vezes estão presentes durante as intervenções, embora houvesse diferenças entre as categorias profissionais a esse respeito. Os procedimentos em que os pais estavam presentes eram aqueles considerados "menos invasivos" (96% dos casos), enquanto apenas 4% estavam presentes naqueles considerados "mais invasivos". Quanto mais velho o profissional, a presença dos pais foi considerada menos necessária. Conclusão: as atitudes em relação à presença dos pais durante o procedimento pediátrico invasivo são influenciadas pela categoria profissional, a idade do prestador de serviço de saúde e a invasividade do procedimento.

Resumen Objetivo: la atención centrada en la familia durante procedimientos invasivos ha sido respaldada por muchas organizaciones profesionales dedicadas al cuidado de la salud. El objetivo de este estudio fue evaluar las actitudes de los profesionales de la salud con respecto a la presencia de los padres durante los procedimientos invasivos realizados en niños. Método: a los prestadores de atención médica en Pediatría (divididos en categorías profesionales y rangos de edad) de uno de los hospitales más importantes de España se les solicitó que respondieran un cuestionario y redactaran comentarios de texto libre. Resultados: un total de 227 profesionales respondieron la encuesta. En sus respuestas, la mayoría (72%) de los participantes informó que, en ocasiones, los padres están presentes durante las intervenciones, aunque se registraron diferencias entre las distintas categorías profesionales al respecto. Los procedimientos en los que los padres estuvieron presentes se consideraron como "menos invasivos" (96% de los casos), mientras que solamente el 4% estuvo presente en los considerados "más invasivos". A mayor edad de los profesionales, menos necesaria se consideró la presencia de los padres. Conclusión: las actitudes con respecto a la presencia de los padres durante procedimientos pediátricos invasivos se vieron influenciadas por la categoría profesional, la edad del prestador de salud y la invasividad de los procedimientos.

Colostrum Features of Active and Recovered COVID-19 Patients Revealed Using Next-Generation Proteomics Technique, SWATH-MS.

Colostrum performs nutritional, anti-inflammatory and anti-infective functions and promotes immune system formation and organ development. The new coronavirus, SARS-CoV-2, has generated concerns about viral transmission through human milk, with a lack of evidence about human milk's protective effects against the infection. This study aimed at analyzing presence of the virus and at identifying the protein expression profile of human colostrum in active and COVID-19-recovered patients. Colostrum samples were collected from women with COVID-19 (n = 3), women recently recovered from the infection (n = 4), and non-infected women (n = 5). The samples were analyzed by means of RT-qPCR to determine presence of the virus and using SWATH-MS for proteomic analysis. Proteomic results were then analyzed using bioinformatic methods. The viral tests were negative for SARS-CoV-2 in the colostrum from COVID-19 patients. The proteomic analysis identified 301 common proteins in all samples analyzed. Nineteen proteins were upregulated and 7 were downregulated in the COVID-19 group versus the control samples, whereas 18 were upregulated and 7 were downregulated when comparing the COVID-19 group to the recovered group. Eleven proteins were biomarkers of active COVID-19 infection. Ten were upregulated: ACTN1, CD36, FAM3B, GPRC5B, IGHA2, IGK, PLTP, RAC1, SDCBP and SERPINF1, and one was downregulated: PSAP. These proteins are mainly related to immunity, inflammatory response and protein transport. In conclusion, the results of this study suggest that colostrum is not a vehicle for mother-to-child SARS-CoV-2 transmission. Moreover, the colostrum's proteome of active and recuperated patients indicate that it could provide immune benefits to infants.

Parental presence during invasive pediatric procedures: what does it depend on?

OBJECTIVE: family-centered care during invasive procedures has been endorsed by many professional health care organizations. The aim of this study was to evaluate the health professionals' attitudes towards parental presence during their child's invasive procedure. METHOD: pediatric healthcare providers (divided in professional categories and range of ages) from one of the Spain's largest hospitals were asked to complete a questionnaire and write free-text comments. RESULTS: 227 responded the survey. Most (72%) participants, in their answers, reported that parents are sometimes present during interventions, although there were differences between professional categories in this respect. The procedures in which the parents were present were those considered "less invasive" (96% of cases), while only 4% were present in those considered "more invasive". The older the professional, the less necessary parental presence was considered. CONCLUSION: the attitudes towards parental presence during pediatric invasive procedure are influenced by the professional category, the age of the healthcare provider and the invasiveness of the procedure.

Analysis of Minor Proteins Present in Breast Milk by Using WGA Lectin.

Breast milk is a complex and dynamic biological fluid and considered an essential source of nutrition in early life. In its composition, the proteins have a relevant biological activity and are related to the multiple benefits demonstrated when compared with artificial milks derived from cow's milk. Understanding human milk composition provides an important tool for health care providers toward the management of infant feeding and the establishment of breastfeeding. In this work, a new technique was developed to increase the knowledge of human milk, because many of the components remain unknown. To isolate minor proteins present in breast milk by using WGA lectin, breast milk was centrifuged to remove cells and separate the fat phase from the serum phase. The serum obtained was separated into two groups: control (n = 3; whole serum sample from mature milk) and WGA lectin (n = 3; sample processed with WGA lectin to isolate glycosylated proteins). The samples were analyzed by high-performance liquid chromatography coupled to mass spectrometry (HPLC/MS). A total of 84 different proteins were identified from all of the samples. In the WGA lectin group, 55 different proteins were isolated, 77% of which had biological functions related to the immune response. Of these proteins, there were eight WGA lectin group exclusives, and two had not previously been described in breast milk (polyubiquitin-B and POTE ankyrin domain family member F). Isolation by WGA lectin is a useful technique to detect minor proteins in breast milk and to identify proteins that could not be observed in whole serum.

Analysis of Uterine Blood Flow in Breeding Sows through the Estrus and Early Diestrus, and after Artificial Insemination.

This study aimed to determine uterine blood flow indices by transabdominal Doppler ultrasound in sows (n = 18) under different conditions: (i) sows after estrus detection (day 0, D0); (ii) sows 2 h after artificial insemination (AI), performed 24 h after detection of estrus (day 1, D1); (iii) sows in early diestrus (day 5, D5). Moreover, three different types of seminal doses were used for AI depending on the ejaculate fraction included (F1: doses containing only the rich fraction of the ejaculate; F2: F1 + the transition fraction between rich and poor fractions; F3: F2 and poor fraction). The statistical analysis revealed significant differences in some indices regarding the period of analysis (D0, D1, and D5). Diastolic velocity and mean velocity showed lower values at D5 in comparison with D0 and D1 (p < 0.01). On the other hand, the pulsatility index and the relationship systolic velocity/diastolic velocity indicated higher values at D5 in comparison with D0 and D1 (p < 0.01). No differences were observed regarding the type of seminal dose used in any of the time points analyzed (p > 0.05). Neither insemination per se nor the type of ejaculate fraction used immediately modified the uterine vascularity, but some indices are affected by the stage of the estrus cycle (estrus vs. early diestrus).

Should All Fractions of the Boar Ejaculate Be Prepared for Insemination Rather Than Using the Sperm Rich Only?

Boar ejaculate is released in several well-characterized fractions, differing in terms of sperm concentration, seminal plasma volume, and composition. However, the inclusion of the last part of the ejaculate for artificial insemination (AI) purposes is still under debate due to its controversial effects. Thus, there is a need to study the potential synergistic impact of the different ejaculate fractions. We aimed to evaluate the effect of accumulative ejaculate fractions on sperm conservation, AI performance, and offspring health. Ejaculates (n = 51) were collected and distributed as follows: F1: sperm-rich fraction; F2: sperm-rich + intermediate fractions; F3: sperm-rich + intermediate + poor fractions. Each group was diluted in a commercial extender, packaged in seminal doses (2000 × 106 sperm/60 mL), and stored at ~16 °C. On day 3 of conservation, sperm were analyzed and used for AI (n = 174). High sperm quality was observed after storage without a significant difference between the groups (p > 0.05). Moreover, no differences were obtained for AI performance (pregnancy and farrowing rates, and litter size; p > 0.05) and offspring health (growth and blood analysis; p > 0.05). Conclusively, the presence of all ejaculate fractions within the seminal doses does not impair the reproductive performance, reporting important economic savings according to the economic model included here.

Evidence of haptoglobin in the porcine female genital tract during oestrous cycle and its effect on in vitro embryo production.

Recent evidence supports involvement of the acute phase protein haptoglobin in numerous events during mammalian reproduction. The present study represents an in-depth investigation of haptoglobin expression and secretion in the porcine oviduct and uterus, and assesses its effect on porcine in vitro embryo production. A systematic study was made of sows in different oestrous stages: late follicular, early luteal and late luteal stages. Relative haptoglobin mRNA abundance was quantified by RT-qPCR. In addition, expression of the protein was analysed by immunohistochemistry and the results were complemented by Western-blot and proteomic analyses of the oviductal and uterine fluids. In vitro porcine fertilization and embryo culture were carried out in the presence of haptoglobin. The results indicate that haptoglobin mRNA expression in the porcine oviduct and uterus is most abundant during the late luteal stage of the oestrous cycle. By means of Western blot and proteomic analyses haptoglobin presence was demonstrated in the oviduct epithelium and in the oviductal and uterine fluids in different stages of the oestrous cycle. The addition of haptoglobin during gamete co-incubation had no effect on sperm penetration, monospermy or efficiency rates; however, compared with the control group, blastocyst development was significantly improved when haptoglobin was present (haptoglobin: 64.50% vs. control: 37.83%; p < 0.05). In conclusion, the presence of haptoglobin in the oviduct and uterus of sows at different stages of the oestrous cycle suggests that it plays an important role in the reproduction process. The addition of haptoglobin during in vitro embryo production improved the blastocyst rates.

Serum acute phase response induced by different vaccination protocols against circovirus type 2 and Mycoplasma hyopneumoniae in piglets.

The purpose of this study was to compare the acute phase reaction (APR) induced by different vaccination protocols used against Porcine Circovirus (PCV) type-2 and Mycoplasma hyopneumoniae (M.hyo), studying two acute phase proteins (APPs) and changes in rectal temperature (RT). In addition, the possible influence of the time of vaccination and breed were analysed. In the first experiment, 40 commercial crossbred piglets were vaccinated, on the day of weaning, with FLEXcombo® (group A, n=20) or Porcilis PCV® and Stellamune® One (group B, n=20). The second experiment was performed on two farms, on which 40 commercial crossbred piglets or 40 Iberian piglets were vaccinated, 7days post-weaning. On each farm one group (A, n=20) was vaccinated with FLEXcombo® and another group (B, n=20) with Porcilis® PCV-M.hyo. Blood samples were taken before, 24h and 48h after vaccination, and RT were recorded before and 8h after vaccination. Significantly higher increases in group B in RT (P<0.01) and APPs concentrations (P<0.01) were recorded at several sampling times after vaccination compared with group A. The vaccines that produced greater increases in RT also produced higher APPs increases but no influence of the day of vaccination or of the breed were found. Therefore, serum APPs concentrations differed according to the vaccine used, which may be useful, along with RT, for choosing the vaccine or protocol that produces APR of lower magnitude.

Optimization of post-cervical artificial insemination in gilts: Effect of cervical relaxation procedures and catheter type.

Post-cervical (pC) artificial insemination (AI) has been successfully developed for application in multiparous sows, although it has proved problematic in gilts. This study analyzes the use of pC-AI in gilts by two experiments. In the first experiment, the efficiency of pC-AI in gilts was evaluated using a multi-ring multiparous catheter (MpC), which led to 23.1% of the gilts being successfully inseminated. In gilts where insemination was not possible using an MpC, two alternatives were applied before a second attempt at insemination: 1) Vetrabutin Chlorhydrate (VC) was intramuscularly injected in order to relax the cervix; or 2) Warm extender (WE) was deposited in the cervix to modify the cervical muscle dynamics. After the application of these treatments, the success rates achieved with the MpC were 34.2% and 23.8% for VC and WE, respectively. No statistically significant differences were found in the reproductive parameters measured [farrowing (%), litter size and fecundity index] between the use of MpC, or the MpC combined with VC or WE, compared with gilts inseminated by cervical AI (control group). In the second experiment, new catheters based on the anatomical characteristics of gilts (GpC) were used, and the rate of successful pC-AI application were compared (experiment 2a): a) MpC: control; b) GpC1: multi-ring catheter of Ø 16 mm and inner cannula of Ø 3.5 mm; c) GpC2: a multi-ring catheter of Ø with an inner cannula of Ø 2.5 mm. The highest rate of successful cannula penetration was reached in the GpC2 group (60.3%) followed by GpC1 (37.0%) and MpC (19.6%) (p < 0.05). There were no differences in the above mentioned reproductive parameters using the three catheters compared with cervical AI method (control group). Moreover, prior cervical AI did not improve subsequent pC-AI application 24 h later (experiment 2b). In conclusion, Vetrabutin Chlorhydrate, warm extender or the new catheters can be considered as useful tools for improving the success rate of pC-AI technique in gilts.

Morphological study of boar sperm during their passage through the female genital tract.

Once deposited in the female tract, sperm face a series of challenges that must be overcome to ensure the presence of an adequate normal sperm population close to the site of fertilization. Our aim was to evaluate the influence of the uterine milieu on boar sperm morphology. In experiment 1, sperm morphology was evaluated in the backflow (60 min after insemination) and within the uterotubal junction (UTJ) (collected ~24 h after insemination) following intrauterine sperm deposition (n = 6) and compared with the morphology of the sperm in the insemination dose. In experiment 2, the influence of the uterine fluid (UF) on sperm morphological modifications was evaluated. For this purpose, ejaculated (n = 4) and epididymal (n = 4) sperm were in vitro incubated with or without UF for 2 and 24 h. In both experiments, sperm were classified as normal, having a cytoplasmic droplet (proximal or distal) or having tail defects. The results of experiment 1 pointed to an increase in morphologically abnormal sperm collected in the backflow (27.70%) and a reduction of the same in the UTJ (2.12%) compared with the insemination dose (17.75%) (P < 0.05). In experiment 2, incubation of ejaculated sperm with UF did not provoke any morphological modifications; however, when epididymal sperm were incubated with UF, a pronounced increase in the percentage of normal sperm was evident after 24 h compared with the initial dose (from 25.77% to 53.58%, P < 0.05), mainly due to distal cytoplasmatic droplet shedding (53.22 vs. 20.20%). In conclusion, almost all the sperm that colonize the UTJ had a normal morphology, with part of the abnormal sperm having been discarded in the backflow and part selected/modified on their way to the oviduct. UF seems to influence cytoplasmic distal droplet removal, as demonstrated previously in seminal plasma.

Morphometry of boar sperm head and flagellum in semen backflow after insemination.

Once deposited in the female reproductive system, sperm begin their competition and undergo a selection to reach the site of fertilization. Little is known about the special characteristics of sperm that reach the oviduct and are able to fertilize, with even less information on the role of sperm dimension and shape in transport and fertilization. Here, we examine whether sperm morphometry could be involved in their journey within the uterus. For this purpose, sperm head dimension (length, width, area, and perimeter) and shape (shape factor, ellipticity, elongation, and regularity), and flagellum length were analyzed in the backflow at different times after insemination (0-15, 16-30, and 31-60 minutes). Sperm morphometry in the backflow was also analyzed taking into account the site of semen deposition (cervical vs. intrauterine). Finally, flagellum length was measured at the uterotubal junction. Sperm analyzed in the backflow were small (head and flagellum) with different head shapes compared with sperm observed in the dose before insemination. The site of deposition influenced head morphometry and tail size both being smaller in the backflow after cervical insemination compared with intrauterine insemination. Mean tail length of sperm collected in the backflow was smaller than that in the insemination dose and at the uterotubal junction. Overall, our results suggest that sperm size may be involved in sperm transport either because of environment or through sperm selection and competence on their way to encounter the female gamete.

Reproductive performance and backflow study in cervical and post-cervical artificial insemination in sows.

The present study was developed to evaluate multiparous sow reproductive performance and backflow in post-cervical artificial insemination (post-CAI) using a reduced number of sperm than in cervical artificial insemination (CAI). The experimental groups were divided into sows inseminated by: 1) cervical artificial insemination (CAI): 3×10(9) spermatozoa/80 ml; 2) post-CAI: 1.5×10(9) spermatozoa/40 ml (post-CAI 1); 3) post-CAI using 1×10(9) spermatozoa/26 ml (post-CAI 2). Post-CAI 1 reproductive parameters were similar to those of post-CAI 2 (except for live born litter size which was greater in post-CAI 1) and better than for the CAI group (p<0.01). In a second experiment the backflow volume, number of sperm, and sperm quality in the backflow were studied in the 3 experimental groups. The % of volume and spermatozoa in the backflow was higher in the CAI group (p<0.05) than post-CAI groups (statistically similar between them). Moreover, the quality parameters (motility, progressive motility, viability, chromatin decondensation and morphology) in backflow semen were identical in all three experimental groups, but differed as regards the original insemination dose incubated inside a colostomy bag (sperm quality control group). The present study shows that the use of post-CAI (either post-CAI 1 or 2) in field conditions can be recommended because the efficiency is similar (in the case of post-CAI 2) or higher (post-CAI 1) than when using the traditional method (CAI), representing a reduction cost.