Molecular epidemiology of paediatric invasive pneumococcal disease in Andalusia, Spain.

This study aimed to assess the impact of the introduction of pneumococcal conjugate vaccine 13 (PCV13) on the molecular epidemiology of invasive pneumococcal disease (IPD) in children from Andalusia. A population-based prospective surveillance study was conducted on IPD in children aged <14 years from Andalusia (2018-2020). Pneumococcal invasive isolates collected between 2006 and 2009 in the two largest tertiary hospitals in Andalusia were used as pre-PCV13 controls for comparison of serotype/genotype distribution. Overall IPD incidence rate was 3.55 cases per 100 000 in 2018; increased non-significantly to 4.20 cases per 100 000 in 2019 and declined in 2020 to 1.69 cases per 100 000 (incidence rate ratio 2020 vs. 2019: 0.40, 95% confidence interval (CI) 0.20-0.89, P = 0.01). Proportion of IPD cases due to PCV13 serotypes in 2018-2020 was 28% (P = 0.0001 for comparison with 2006-2009). Serotypes 24F (15%) and 11A (8.3%) were the most frequently identified non-PCV13 serotypes (NVT) in 2018-2020. Penicillin- and/or ampicillin-resistant clones mostly belonged to clonal complex 156 (serotype 14-ST156 and ST2944 and serotype 11A-ST6521). The proportion of IPD cases caused by PCV13 serotypes declined significantly after the initiation of the PCV13 vaccination programme in 2016. Certain NVT, such as serotypes 24F and 11A, warrant future monitoring in IPD owing to invasive potential and/or antibiotic resistance rates.

Selenomethionine administration decreases the oxidative stress induced by post mortem ischemia in the heart, liver and kidneys of rats.

Selenium is an essential element in human and animal metabolism integrated into the catalytic site of glutathione peroxidase (GPX1), an antioxidant enzyme that protects cells from damage caused by reactive oxygen species (ROS). Oxidative stress refers the imbalance between ROS and antioxidant defense systems. It generates alterations of DNA, proteins and lipid peroxidation. The imbalance occurs particularly during ischemia and lack of postmortem perfusion. This mechanism is of relevance in transplant organs, affecting their survival. The aim of this research is to evaluate the effect of seleno-methionine (SeMet) as a protective agent against postmortem ischemia injury in transplant organs. Wistar rats were orally administered with SeMet. After sacrifice, liver, heart and kidney samples were collected at different postmortem intervals (PMIs). SeMet administration produced a significant increase of Se concentration in the liver (65%, p < 0.001), heart (40%, p < 0.01) and kidneys (45%, p < 0.05). Levels of the oxidative stress marker malondialdehyde (MDA) decreased significantly compared to control in the heart (0.21 ± 0.04 vs. 0.12 ± 0.02 mmol g-1) and kidneys (0.41 ± 0.02 vs. 0.24 ± 0.03 mmol g-1) in a PMI of 1-12 h (p < 0.01). After SeMet administration for 21 days, a significant increase in GPX1 activity was observed in the liver (80%, p < 0.001), kidneys (74%, p < 0.01) and heart (35%, p < 0.05). SeMet administration to rats significantly decreased the oxidative stress in the heart, liver and kidneys of rats generated by postmortem ischemia.

Differential estrogen-related responses in myofiber cross-sectional area of pelvic floor muscles in female rats.

OBJECTIVE: To determine the role of estrogens in myofiber cross-sectional area (CSA) of the pubococcyegeus (Pcm) and iliococcygeus muscles (Icm). METHODS: In Experiment 1, we excised the Pcm and Icm during the metestrus and proestrus stages of the estrous cycle to measure the myofiber CSA. In Experiment 2, we allocated other rats into the following groups: sham (Sh), ovariectomized (OVX), OVX plus 1,4,6-androstatriene-3,17-dione (ATD; OVX + ATD), an aromatase inhibitor, and OVX plus estradiol benzoate (OVX + EB). We carried out appropriate statistical tests to determine significant differences (p ≤ 0.05) in variables measured for both Experiments. RESULTS: The Pcm myofiber CSA at proestrus was higher than at metestrus, while the Icm myofiber CSA did not change. Ovariectomy increased the Pcm myofiber CSA, which was exacerbated with the ATD administration. The EB supplementation successfully reversed the ovariectomy-induced enlargement of the CSA. No significant changes were detected for the Icm myofiber CSA. CONCLUSIONS: Fluctuating ovarian steroid levels at the estrus cycle significantly influence the CSA myofiber of the Pcm but not that of the Icm. Estrogen actions, having a gonadal or extragonadal origin, influence importantly the CSA of the Pcm.

Bladder and urethral dysfunction in multiparous and mature rabbits correlates with abnormal activity of pubococcygeus and bulbospongiosus muscles.

AIMS: To evaluate the combined effect of age and multiparity on the micturition reflex, including pelvic floor muscle activation. METHODS: Young and mature nulliparous rabbits were compared to young and mature multiparas (n = 6 per group). Cystometrograms and urethral pressure (UP) were performed while simultaneously recording the electromyogram (EMG) activity of the pubococcygeus and bulbospongiosus muscles to establish their functional correlation to urological function. RESULTS: Multiparity and age significantly influence the bladder and UP affecting the voiding efficiency and intercontraction interval. Such interaction also reduced the UP threshold, timing, and duration. Other bladder and urethral variables were predominantly affected only by age. Urodynamic alterations correlated with abnormal patterns or absent EMG activity of the pubococcygeus and bulbospongiosus muscles. CONCLUSIONS: The present findings strongly suggest that multiparity and age affects specific pelvic floor muscle reflex activation during micturition, and may contribute to alterations in bladder and urethral function. This data broadens our understanding of the critical role of the appropriate activity of the individual pelvic floor muscles in micturition.

Arsenic species uptake and translocation in Elodea canadensis.

The capacity of Elodea canadensis to phytofiltrate arsenic species from water was evaluated. Plants were adapted to tap water and supplemented with 15 and 250 µg L-1 of As. Inorganic arsenic species (As III, As V), and organic arsenic compounds: monomethylarsonate (MMA) and dimethylarsinate (DMA) were analyzed. Sampling was carried out at different times after exposure in culture water and plant organs. Plants exposed to 15 µg L-1 of As concentration showed no significant difference on As concentration (95% confidence level) in their organs compared to controls. When plants were exposed to 250 µg L-1 of As concentration, a significant increase of As concentration in plant organs was observed. After 1 h exposure, plants reduce 63.16% the As concentration in the culture water, with a bioaccumulation factor (BF) of 4.3. Under these conditions, E. canadensis accumulate As V in roots and do not translocate it to stems (transfer factor <1). MMA was determined in stems and leaves. E. canadensis effectively phytofiltrate As from tap water of a city located in an arsenic endemic area from concentrations of 36 µg L-1 to undetectable levels (10 ng L-1).

Respiratory Syncytial Virus Seasonality: A Global Overview.

Respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory infections in children. By the age of 1 year, 60%-70% of children have been infected by RSV. In addition, early-life RSV infection is associated with the development of recurrent wheezing and asthma in infancy and childhood. The need for precise epidemiologic data regarding RSV as a worldwide pathogen has been growing steadily as novel RSV therapeutics are reaching the final stages of development. To optimize the prevention, diagnosis, and treatment of RSV infection in a timely manner, knowledge about the differences in the timing of the RSV epidemics worldwide is needed. Previous analyses, based on literature reviews of individual reports obtained from medical databases, have failed to provide global country seasonality patterns. Until recently, only certain countries have been recording RSV incidence through their own surveillance systems. This analysis was based on national RSV surveillance reports and medical databases from 27 countries worldwide. This is the first study to use original-source, high-quality surveillance data to establish a global, robust, and homogeneous report on global country-specific RSV seasonality.

The metabolic effects of mercury during the biological cycle of vines (Vitis vinifera).

Mercury (Hg) is a major environmental pollutant that can be disposed to the environment by human activities, reaching crops like vineyards during irrigation with contaminated waters. A 2-year study was performed to monitor Hg variations during reproductive and vegetative stages of vines after Hg supplementation. Variations were focused on total Hg concentration, the molecular weight of Hg fractions and Hg-proteins associations in roots, stems and leaves. Total Hg concentrations increased during reproductive stages and decreased during vegetative stages. Variations in length of these stages were observed, according to an extension of the vegetative period. Six months post Hg administration, in roots, stems and leaves, initial Hg proteic fractions of 200 kDa were catabolized to 66 kDa fractions according to a transition from reproductive to vegetative stages. However, 24 months after Hg supplementation, the 66 kDa Hg proteic fraction was continuously determined in a prolonged senescence. Accordingly, the identified proteins associated to Hg show catabolic functions such as endopeptidases, hydrolases, glucosidases and nucleosidases. Stress associated proteins, like peroxidase and chitinase were also found associated to Hg. During the reproductive periods of vines, Hg was associated to membrane proteins, such as ATPases and lipid transfer proteins, especially in roots where Hg is absorbed.

Actor-specific contributions to the deforestation slowdown in the Brazilian Amazon.

Annual deforestation rates in the Brazilian Amazon fell by 77% between 2004 and 2011, yet have stabilized since 2009 at 5,000-7,000 km(2). We provide the first submunicipality assessment, to our knowledge, of actor-specific contributions to the deforestation slowdown by linking agricultural census and remote-sensing data on deforestation and forest degradation. Almost half (36,158 km(2)) of the deforestation between 2004 and 2011 occurred in areas dominated by larger properties (>500 ha), whereas only 12% (9,720 km(2)) occurred in areas dominated by smallholder properties (<100 ha). In addition, forests in areas dominated by smallholders tend to be less fragmented and less degraded. However, although annual deforestation rates fell during this period by 68-85% for all actors, the contribution of the largest landholders (>2,500 ha) to annual deforestation decreased over time (63% decrease between 2005 and 2011), whereas that of smallholders went up by a similar amount (69%) during the same period. In addition, the deforestation share attributable to remote areas increased by 88% between 2009 and 2011. These observations are consistent across the Brazilian Amazon, regardless of geographical differences in actor dominance or socioenvironmental context. Our findings suggest that deforestation policies to date, which have been particularly focused on command and control measures on larger properties in deforestation hotspots, may be increasingly limited in their effectiveness and fail to address all actors equally. Further reductions in deforestation are likely to be increasingly costly and require actor-tailored approaches, including better monitoring to detect small-scale deforestation and a shift toward more incentive-based conservation policies.

Coital Urinary Incontinence Induced by Impairment of the Dorsal Nerve of the Clitoris in Rats.

PURPOSE: We determined the effect of chronic bilateral neurectomy of the dorsal nerve of the clitoris on urinary parameters and sexual behavior of conscious female rats. MATERIALS AND METHODS: A total of 18 anesthetized virgin female Wistar rats were used in this study, including 11 that underwent bilateral neurectomy of the dorsal nerve of the clitoris and 7 that underwent sham surgery. Urinary parameters were determined in awake animals preoperatively, and 3 and 10 days postoperatively. Sexual behavior was tested 14 days postoperatively to determine whether the females expelled urine during sexual encounters. After male ejaculation the females were anesthetized with urethane to record external urethral sphincter electromyogram activity in response to clitoris, perigenital skin and vaginal stimulation. Neurectomy was corroborated anatomically. RESULTS: Sham surgery did not significantly modify urinary parameter values. However, bilateral neurectomy of the dorsal nerve of the clitoris significantly increased voiding frequency and voiding duration (p <0.05). It did not significantly affect the flow rate, voided volume or voiding interval. Of females that underwent bilateral neurectomy of the dorsal nerve of the clitoris 67% expelled urine just after male ejaculation. CONCLUSIONS: These results suggest that the pudendal nerve is an important neural pathway in the convergence and crosstalk of female urogenital neural circuits, and genital deafferentation may be a causal factor of coital urinary incontinence. Rats with bilateral transection of the dorsal nerve of the clitoris may serve as an animal model of coital incontinence.

Liquid chromatography coupled to molecular fluorescence with postcolumn UV sensitization for thimerosal and derivative compounds monitoring in environmental samples.

A HPLC coupled with molecular fluorescence (MF) spectrometry method for determination of thimerosal (THM, sodium ethylmercurythiosalicylate, C9 H9 HgNaO2 S), and derivatives is proposed. A sensitization of MF was provoked by UV irradiation of analytes in a home-made photoreactor that served as interface between the LC column and MF spectrometer. This method is applied to determination of THM, ethyl mercury, and thiosalicylic acid in samples of pharmaceutical industry effluents, and waters of La Carolina and Jáchal rivers situated in the center-west side of San Luis city and in the east of San Juan city (Middle West, Argentine) where the effluents are dumped. The LODs calculated on basis of 3σ criterion were 1.8, 5, and 0.05 µmol/L for THM, ethyl mercury, and for thiosalicylic acid, respectively.

Activity of the external urethral sphincter evoked by genital stimulation in male rats.

AIMS: To determine whether the external urethral sphincter (EUS) fasciculi of male rats respond to the mechanical stimulation of genital structures and to characterize the pattern of the electromyographic (EMG) activity of the three regions of the EUS: the cranial (CrEUS), the medial (MeEUS) and the caudal (CaEUS). METHODS: Electromyographic signals were recorded from the CrEUS, MeEUS and CaEUS regions of the male rat's EUS, before, during and after the mechanical stimulation of the urogenital structures. RESULTS: The CrEUS, MeEUS and CaEUS regions responded when brushing and squeezing the foreskin and glans as well as to penile and prostatic urethral distension. The CaEUS EMG amplitude (P < 0.01) and frequency (P < 0.05) were lower in comparison to the CrEUS and MeEUS responses to the mechanical stimulation. In addition, the CaEUS was characterized by a short or no afterdischarge. In contrast, the CrEUS and MeEUS responded by presenting a long discharge after the penile or prostatic urethral distension. CONCLUSIONS: The activity of the EUS is modulated by both, cutaneous and visceral genitourinary stimuli, with motor units being activated by mechanoreceptors located in the foreskin, glans, bladder, and urethra. The CrEUS, MeEUS and CaEUS have differential EMG patterns, indicating that the EUS consists of three anatomically and functionally different regions. Precise coordination in the muscular activity of these regions may be crucial for the control of male expulsive urethral functions, i.e., during voiding and ejaculation. Neurourol. Urodynam. 35:914-919, 2016. © 2015 Wiley Periodicals, Inc.

Differential damage and repair responses of pubococcygeus and bulbospongiosus muscles in multiparous rabbits.

AIM: To determine the extent of damage and regeneration associated with multiparity on the pubococcygeus and bulbospongiosus muscles. METHODS: Age-matched virgin nulliparous and multiparous rabbits that were killed at days 3 and 20 after the fourth delivery were used to harvest pubococcygeus and bulbospongiosus muscles. The activity of the lysosomal enzyme ß-glucuronidase was used as a muscle damage indicator. The number of immunoreactive myofiber-associated nuclei anti-Pax7, -MyoD, and -myogenin, as well as the anti-desmin immunoreactive area were measured in muscle sections to estimate some regenerative stages. Significant differences were considered at a P ≤ 0.05. RESULTS: The ß-glucuronidase activity was increased at postpartum day 20 in the pubococcygeus muscle. This variable was unaltered in the bulbospongiosus muscles of multiparas regardless of the postpartum day on which this was measured. The number of immunoreactive nuclei anti-Pax7 in the pubococcygeus muscle was similar between nulliparas and multiparas, whilst those of anti-MyoD and anti-myogenin were increased at postpartum days 3 and 20. The same was true for these latter three markers evaluated in the bulbospongiosus muscles, supporting an ongoing regeneration. The desmin-positive percentage of muscle area per field was increased at postpartum day 20 in the pubococcygeus muscle, whilst such an increment was seen at postpartum days 3 and 20 in the bulbospongiosus muscles. CONCLUSIONS: Damage and regeneration of the pubococcygeus and bulbospongiosus muscles are differently influenced by multiparity in rabbits. This could rely on the anatomical location, metabolism, myofiber composition, and muscle exertion during pregnancy and/or the delivery of each muscle.

Liver failure secondary to poisoning by a homemade product made of star and green anise in a 4-month-old infant.

Intoxications in pediatric age represent a frequent cause of visit to the hospital emergency unit. Herb-made products can be toxic for the infant. The neurotoxic properties of the star anise (Illicium verum) have been widely described, although it is a classic product used to treat the infantile colic. Hepatic failure due to the consumption of anise herb elaborated infusions is presented as an exceptional finding in our environment. A case of a 4-month-old infant with hypertransaminasemia, severe coagulopathy, non ketotic hypoglycemia, moderated metabolic acidosis and neurologic symptoms such as seizures and nistagmus is described. After discarding infectious, metabolic and autoimmune etiology and through a meticulous anamnesis, the family referred having administered in the last two months a daily star anise and green anise (Pimpinella anisum) infusion to the patient. It is important to emphasize the serious risk of administering homemade herb infusions to infants.

Prognosis of atypical teratoid rhabdoid tumors (AT/RT) treated with multimodal therapy protocols. Report of our series.

OBJECTIVE: Atypical teratoid rhabdoid tumors (AT/RT) of the central nervous system are rare, very aggressive embryological tumors, typically diagnosed in young patients and having a low survival rate after diagnosis. The aim of this study was to emphasize, based on the latest results in the literature, the need for protocols for multidisciplinary treatment in these patients. MATERIAL AND METHODS: We report our series of 3 cases treated, diagnosed and followed up between 2009 and 2014. They were treated with multimodal therapy protocols (Rhabdoid SIOP-2007 and European Rhabdoid Registry EU-RHAB-2010). In addition, we carried out a literature review. RESULTS: Two of our 3 cases (supratentorial and spinal tumors) did not show any progression of the disease after long follow-up, in contrast with most of the cases available in the literature. The second patient had a shorter survival. CONCLUSIONS: Patient age at the time of diagnosis, supratentorial location of the mass and fewer complications with adjuvant treatments seem to be factors yielding good prognosis for AT/RT tumors. In agreement with the latest international protocols, multidisciplinary treatment is the ideal treatment, consisting of radiotherapy and chemotherapy after complete tumor resection.

Aromatase expression is linked to estrogenic sensitivity of periurethral muscles in female rabbits.

Beyond its role in the conversion of androgens to estrogens, the expression of aromatase could influence on the estrogenic signalling in targeted tissues. Considering the well-defined biochemical and physiological differences between the pubococcygeus (Pcm) and bulbospongiosus (Bsm) muscles in female rabbits, it is presently hypothesized that the aromatase expression is differentially linked to the estrogen sensitivity of each muscle. To this end, serum estradiol levels and the aromatase expression, presence of ERα and ERß and morphometry were evaluated in the Pcm and Bsm of female rabbits allocated in control, ovariectomized (OVX) and OVX treated with estradiol benzoate (OVX + EB) groups. Aromatase expression was high in the Pcm. Independently to serum estradiol, ovariectomy increased aromatase expression in the Pcm while decreased it in the Bsm. The EB treatment avoided the effect of ovariectomy only in the Pcm. The number of immunoreactive nuclei anti-ERα and anti-ERß was high in the Pcm of OVX and OVX + EB rabbits, while those in the Bsm remained unchanged. The number of peripheral nuclei per fibre and the cross-sectional area-to-myonucleus ratio were modified only in the Pcm. Our findings support aromatase expression in the Pcm, and Bsm of rabbits is differentially linked to estrogenic sensitivity of each muscle.

Characterization of Hg-phytochelatins complexes in vines (Vitis vinifera cv Malbec) as defense mechanism against metal stress.

An approach to understand vines (Vitis vinifera) defense mechanism against heavy metal stress by isolation and determination of Hg-phytochelatins (PCs) complexes was performed. PCs are important molecules involved in the control of metal concentration in plants. PCs complex toxic metals through -SH groups and stores them inside cells vacuole avoiding any toxic effect of free metals in the cytosol. The Hg-PCs identification was achieved by determination of Hg and S as hetero-tagged atoms. A method involving two-dimensional chromatographic analysis coupled to atomic spectrometry and confirmation by tandem mass spectrometry is proposed. An approach involving size exclusion chromatography coupled to inductively coupled plasma mass spectrometry on roots, stems, and leaves extracts describing Hg distribution according to molecular weight and sulfur associations is proposed for the first time. Medium-low molecular weight Hg-S associations of 29-100 kDa were found, suggesting PCs presence. A second approach employing reversed-phase chromatography coupled to atomic fluorescence spectrometry analysis allowed the determination of Hg-PCs complexes within the mentioned fractions. Chromatograms showed Hg-PC2, Hg-PC3 and Hg-PC4 presence only in roots. Hg-PCs presence in roots was confirmed by ESI-MS/MS analysis.

Molecular epidemiology of pneumococcal carriage in children from Seville, following implementation of the PCV13 immunization program in Andalusia, Spain.

INTRODUCTION: The 13-valent pneumococcal conjugate vaccine (PCV13) universal vaccination programme was introduced in December 2016 in Andalusia. METHODS: A cross-sectional study was conducted on the molecular epidemiology of pneumococcal nasopharyngeal colonization. A total of 397 healthy children were recruited from primary healthcare centres in Seville for the periods 1/4/2018 to 28/2/2020 and 1/11/2021 to 28/2/2022 (PCV13 period). Data from a previous carriage study conducted among healthy and sick children from 1/01/2006 to 30/06/2008 (PCV7 period), were used for comparison of serotype/genotype distributions and antibiotic resistance rates. RESULTS: Overall, 76 (19%) children were colonized with S. pneumoniae during the PCV13 period and there were information available from 154 isolates collected during the PCV7 period. Colonization with PCV13 serotypes declined significantly in the PCV13 period compared with historical controls (11% vs 38%, p = 0.0001), being serotypes 19F (8%), 3 (1%) and 6B (1%) the only circulating vaccine types. Serotypes 15B/C and 11A were the most frequently identified non-PCV13 serotypes during the PCV13 period (14% and 11%, respectively); the later one increased significantly between time periods (p = 0.04). Serotype 11A was exclusively associated in the PCV13 period with ampicillin-resistant variants of the Spain9V-ST156 clone (ST6521 and genetically related ST14698), not detected in the preceding period. CONCLUSIONS: There was a residual circulation of vaccine types following PCV13 introduction, apart from serotype 19F. Serotype 11A increased between PCV13 and PCV7 periods due to emergence and clonal expansion of ampicillin-resistant genotype ST6521.

Inner capillary diameter of hypothalamic paraventricular nucleus of female rat increases during lactation.

BACKGROUND: The role of the endothelial cell (EC) in blood flow regulation within the central nervous system has been little studied. Here, we explored EC participation in morphological changes of the anterior hypothalamic paraventricular nucleus (PVN) microvasculature of female rats at two reproductive stages with different metabolic demand (virginity and lactation). We measured the inner capillary diameter (ICD) of 800 capillaries from either the magnocellular or parvocellular regions. The space occupied by neural (somas, dendrites and axons) and glial, but excluding vascular elements of the neurovascular compartment was also measured in 100-µm2 sample fields of both PVN subdivisions. RESULTS: The PVN of both groups of animals showed ICDs that ranged from 3 to 10 microns. The virgin group presented mostly capillaries with small ICD, whereas the lactating females exhibited a significant increment in the percentage of capillaries with larger ICD. The space occupied by the neural and glial elements of the neurovascular compartment did not show changes with lactation. CONCLUSIONS: Our findings suggest that during lactation the microvasculature of the PVN of female rats undergoes dynamic, transitory changes in blood flow as represented by an increment in the ICD through a self-cytoplasmic volume modification reflected by EC changes. A model of this process is proposed.

Risk assessment on irrigation of Vitis vinifera L. cv Malbec with Hg contaminated waters.

Concerns regard watering crops with Hg contaminated waters have arisen worldwide recently. In these sense Hg uptake by Vitis vinifera L. cv. Malbec was evaluated under greenhouse conditions by the administration of Hg(2+) for 4 days through irrigation water (short-term administration). Vines uptake Hg translocating it from roots through stems to leaves. Roots accumulated the higher Hg concentration. Hg in stems and leaves was accumulated mostly as organic Hg, bind to different moieties. Size exclusion chromatography (SEC) and ion pair chromatography (IPC) were employed to reach insights into these ligands. Hg is distributed mainly in high molecular weight fractions of 669 kDa in vine plants. In stems and leaves, Hg-S associations were found in 669 and 66 kDa fractions. Hg-S association at 66 kDa suggests a possible protein or peptide binding affecting vines normal physiology. Since Hg contamination through organomercurials is more harmful than Hg(2+) itself, methyl mercury, dimethyl mercury, and phenyl mercury, more toxic Hg species were evaluated with negative results.