Invasive meningococcal disease in Chile seven years after ACWY conjugate vaccine introduction.

BACKGROUND: A serogroup W (MenW) outbreak in Chile prompted a meningococcal vaccination campaign using tetravalent meningococcal-conjugate vaccines (MCV-ACWY) in children since 2012, followed by its introduction into the National Immunization Program (NIP) in toddlers from 2014. Direct protection was observed, but no indirect effects in other age-groups were evidenced. The aim of this study was to describe invasive meningococcal disease (IMD) cases in Chile between 2009 and 2019, and its trend after the introduction of MCV-ACWYs. METHODS: IMD cases, cumulative incidence per 100,000 inhabitants, CFR, and vaccination uptake were described. Data were obtained from the Public Health Institute and NIP. RESULTS: Overall-IMD cases increased in 2009-2014 period, followed by a decline in 2015-2019, focused in infants, children <5 years and people ≥60 years. Serogroup B (MenB) and MenW alternate its predominance. Median overall incidence was 0.6/100,000, increasing from 0.6/100,000 in 2009 to 0.8/100,000 in 2014, later decreasing to 0.4/100,000 in 2019. Median incidences for MenB, serogroup C (MenC) and Y (MenY) were 0.25/100,000, <0.01/100,000 and <0.01/100,000, respectively. Median MenW incidence was 0.53/100,000, increasing from 0.01/100,000 in 2009 to 0.56/100,000 in 2014, followed by a constant decline to 0.12 in 2019. Infants, children <5 years and adults ≥60 years were affected the most, with median incidences of 9.7, 0.9 and 0.93, decreasing to 1.3, 0.1 and 0.1/100,000 in 2019, respectively. Median overall-CFR was 19%, 7.5% for MenB and 24.5% for MenW. Median MCV-ACWY uptake was 93% CONCLUSION: Overall-IMD, MenW cases and incidence declined since 2015 after the MCV-ACWY introduction, while MenB, MenC and MenY have been stable. MenW incidence declined in all age groups, including non-immunized infants and people >60 years. Further analysis and a longer period of observation are needed to have a more robust conclusion about this epidemiological trend. By 2019, CFR remains high.

Meningococcal invasive disease by serogroup W and use of ACWY conjugate vaccines as control strategy in Chile.

BACKGROUND: Serogroup causing invasive meningococcal disease (IMD) can change abruptly, as it occurred in Chile when serogroup predominance switched from MenB to MenW in 2012. As a response, a national vaccination strategy was implemented since 2012 using tetravalent meningococcal-conjugate vaccines (MCV-ACWY) in children 9 months through 4 years of age. The aim of this study was to describe IMD cases by MenW in Chile 2009-2016, and to analyse its trend after the introduction of MCV-ACWY. METHODS: Descriptive study of IMD cases in Chile, period 2009-2016. Cumulative incidence and mortality rate per 100,000 inhabitants, and case fatality rate (CRF) were used for descriptive analysis. Linear regression was used for post-intervention trend analysis. RESULTS: In 2012, MenW, mainly ST-11 cc, became predominant. MenW incidence rose from 0.01/100,000 inhabitants in 2009 to a maximum of 0.6/100,000 in 2015. Infants and adults 80 years of age and older were mostly affected, with an incidence peak of 9.7/100,000 and 1.6/100,000, respectively, in 2015. In the group of children from 1 to 4 years of age MenW incidence declined from 1.3/100,000 in 2012 to 0.1/100,000 in 2016, a 92.3% reduction after vaccination implementation. In the same period and age-cohort, CFR decreased from 23% to 0%. High mortality rates concentrated in infants and adults 80 years of age and over. CONCLUSION: MenW became predominant in Chile since 2012. IMD cases increased steadily from 2009 to 2016, with higher incidence, CFR and mortality concentrating in infants and people 80 years of age and older. MCV-ACWY provided direct protection against MenW, reducing its incidence after mass meningococcal vaccine implementation. Indirect effects of vaccination are not yet observed.

Combined administration of a gonadotropin-releasing hormone antagonist and testosterone in men induces reversible azoospermia without loss of libido.

GnRH antagonists suppress pituitary and gonadal function by competing with endogenous GnRH for binding to receptors on pituitary gonadotrophs. We studied the effects of GnRH antagonist administration to men in a protocol simulating a likely male contraceptive regimen combined with a low dose of testosterone. The GnRH antagonist Nal-Glu was given daily (10 mg, sc) for 20 weeks to eight normal men, and a low dose of testosterone enanthate (25 mg, sc) was given every week. Sperm counts started declining during week 4, and complete azoospermia was reached within 6-12 weeks in six of the eight subjects. Subjects 7 and 8, whose sperm counts and serum gonadotropin levels were not suppressed after 10 weeks, were given 20 mg Nal-Glu starting at week 10. One became azoospermic at week 16, while the other's total sperm counts continued declining and reached a nadir of 1.4 million by week 20. Sperm motility and viability in this subject were completely suppressed after week 14. Sperm counts returned to baseline levels 12-14 weeks after the end of Nal-Glu administration. The mean serum LH level of the first six subjects decreased from 3 +/- 03. U/L at baseline to less than 0.1 U/L until week 20, and then levels returned to baseline. FSH levels similarly decreased from a combined mean of 3.6 +/- 0.9 U/L at baseline to below 0.3 U/L after 4 weeks of Nal-Glu administration. Serum mean testosterone levels between weekly injections of testosterone enanthate ranged from 27.4 +/- 5.9 to 4.8 +/- 1.4 nmol/L, but remained in the hypogonadal range (less than 10 nmol/L) for 4 of the 7 days. None of the subjects, however, complained of decreased libido or potency, as assessed by a questionnaire. No systemic or significant local side-effects were observed, other than a minimal reaction at the injection site. These data suggest that complete sustained azoospermia can be achieved in man, without loss of libido, by chronic administration of a GnRH antagonist plus testosterone.

Distribution of angiotensin IV binding sites (AT4 receptor) in the human forebrain, midbrain and pons as visualised by in vitro receptor autoradiography.

Angiotensin IV and other AT4 receptor agonists, improve memory retention and retrieval in the passive avoidance and swim maze learning paradigms. Angiotensin IV binding sites (also known as the AT4 receptors) are widely distributed in guinea pig and monkey (Macaca fascicularis) brains where high densities of the binding sites have been detected in the hippocampus, neocortex and motor nuclei. However, the distribution of the binding sites in the human brain is not known. We have recently localised the angiotensin IV binding sites (AT4 receptors) in post-mortem human brain using iodinated Nle-angiotensin IV, a higher affinity and more stable analogue of angiotensin IV. This radioligand bound with relatively high affinity and specificity to angiotensin IV binding sites. In competition studies on consecutive sections through the prefrontal cortex and claustrum, angiotensin IV, Nle-angiotensin IV and LVV-hemorphin 7 competed for the binding of 125I[Nle]-angiotensin IV with nanomolar affinities. Angiotensin II and the AT1 and AT2 receptor antagonists were ineffective in competing for the binding at concentrations of up to 10 microM. We found high densities of 125I[Nle]-angiotensin IV binding sites throughout the cerebral cortex including the insular, entorhinal, prefrontal and cingulate cortices. Very high densities of the binding sites were observed in the claustrum, choroid plexus, hippocampus and pontine nucleus. Some thalamic nuclei displayed high densities of binding including the anteroprincipal, ventroanterior, anteromedial, medial dorsal and ventrolateral nuclei. The caudate nucleus, putamen, many amygdaloid nuclei and the red nucleus all displayed moderate densities of binding with a higher level detected in the substantia nigra pars compacta. In the hypothalamus, high densities binding sites were found in the ventromedial nucleus with lower levels in the dorsomedial and paraventricular nuclei. The distribution of 125I[Nle]-angiotensin IV binding sites in the human brain is similar to that found in other species and supports multiple roles for the binding sites in the central nervous system, including facilitation of memory retention and retrieval.

Sperm motion parameters after suppression of spermatogenesis with a gonadotropin-releasing hormone antagonist plus testosterone supplementation.

OBJECTIVE: To evaluate whether the chronic administration of a GnRH antagonist supplemented with T enanthate affects sperm motion parameters. DESIGN: Prospective study. SETTING: Academic medical research environment. PATIENTS/PARTICIPANTS: Six normospermic men of reproductive age. INTERVENTION: A 20-week treatment included the administration of 10 mg of GnRH antagonist every day and 25 mg of T enanthate once a week. MAIN OUTCOME MEASURE: Computerized sperm motion analysis on each participant every 2 weeks for a period of 11 months. RESULTS: Sperm concentration decreased after 4 weeks of the GnRH antagonist administration, accompanied by a reduction of sperm motility from an initial mean value of 56% to 34% on treatment week 4 and 21% on treatment week 6. In contrast, sperm velocity, linearity, amplitude of the lateral head displacement, and beat cross-frequency values did not significantly change in spite of reduced overall sperm concentration and motility. Azoospermia was reached within 6 to 12 weeks of the GnRH administration, and it was sustained during the treatment period. During recovery of spermatogenesis, sperm movement parameters returned to initial values earlier than sperm concentration and normal morphology. All participants recovered initial semen parameters 20 weeks after the end of treatment. CONCLUSION: The administration of a GnRH antagonist supplemented with T enanthate successfully induced reversible suppression of spermatogenesis. The sperm motion characteristics appeared to be the last parameters to decline and the first to return to initial values after the treatment.

In vitro deleterious effect of hypoxanthine in Ham's Nutrient Mixture F-10 culture medium on human oocyte fertilization and early embryonic development.

OBJECTIVE: To determine whether hypoxanthine in Ham's Nutrient Mixture F-10 (GIBCO, Grand Island, NY) culture medium affects murine embryo development or the outcome of patients undergoing IVF-ET. DESIGN: For the two-cell embryo bioassay, embryos from each female were equally distributed and incubated in Ham's F-10 with or without hypoxanthine supplementation for up to 72 hours. To assess the effect of hypoxanthine in human IVF-ET, oocytes, sperm, and embryos were cultured in Ham's F-10 medium with or without hypoxanthine. Fertilization and embryo cleavage were assessed at 18 and 40 hours, respectively, after insemination. SETTING: University medical research laboratory. PATIENTS, PARTICIPANTS: Nine couples undergoing IVF-ET. RESULTS: Two-cell mouse embryos incubated for 24 hours without hypoxanthine developed 40% to morula, compared with 6.5% for the hypoxanthine group. At 72 hours, 99.5% of the embryos without hypoxanthine reached the expanded blastocyst stage with 65% of them exhibiting hatching, compared with 72% and 19.5%, respectively, for the group with hypoxanthine. Human oocytes cultured in Ham's F-10 without hypoxanthine showed higher fertilization rates than the group incubated in the presence of hypoxanthine (69% versus 53%). The proportion of cleaved embryos was not different between the two culture media; however, the rate of embryos cleaving without cytoplasmic fragments was significantly higher in the group without hypoxanthine (75% versus 35%). CONCLUSION: Ham's F-10 medium containing hypoxanthine significantly reduced embryo development in the two-cell mouse embryo bioassay. Hypoxanthine in culture medium for IVF-ET may have a deleterious effect on human gametes, leading to decreased fertilization and increased incidence of cytoplasmic fragments in the conceptuses.

The efficacy of cryopreserved hamster ova in the sperm penetration assay.

To solve the logistical problems of the sperm penetration assay (SPA) to provide just a sufficient number of hamster ova exactly when they are needed, a new method to cryopreserve the ova has been devised. The ova, suspended in a 1.5 M solution of propylene glycol as a cryoprotectant in an isotonic salt solution, were frozen in 1/4 mL plastic straws. Included in each straw was a sucrose solution, isosmotic to the propylene glycol solution, to serve as an osmotic buffer during dilution of the cryoprotectant out of the ova. This one-step method of dilution permitted the ova to be recovered and diluted out of the cryoprotectant within the straw in which they had been originally frozen. A total of 547 cryopreserved ova were thawed, 504 (92.1%) of which were morphologically normal after they had been incubated at 37 degrees C for 3 hours. After removal of the zonae, the frozen-thawed ova were compared with fresh, control ova in SPAs of donor and patient semen that had been capacitated in TEST-yolk buffer. The percent penetration and penetration index of fresh versus cryopreserved ova did not differ significantly for either donor or patient semen.

Coculture of mouse embryos with cells isolated from the human ovarian follicle, oviduct, and uterine endometrium.

OBJECTIVE: To examine the specificity of somatic cell support by comparing embryonic development during long-term in vitro coculture with feeder cells derived from the human ovarian follicle, oviduct, and endometrium. DESIGN: Comparative study of murine embryo development and degeneration during 6 days of in vitro coculture. RESULTS: All feeder-cell cultures were beneficial to embryonic development and viability. Few differences were observed between feeder cell types (epithelial or fibroblastic) or cell origin (ovarian follicle, oviductal, or endometrial). Embryos developed to the eight-cell stage in 24 hours whether in coculture (83.6% to 100%) or in media alone (85.2%); however, further development in media alone decreased compared with coculture (15.6% versus 63.4% to 87.7%, plating) and embryo degeneration increased (67.9% versus 5.5% to 19.4%) after 6 days. CONCLUSIONS: [1] Coculture of embryos with human reproductive tract cells is beneficial to embryonic development and viability. [2] Human somatic cell support of murine embryos during long-term in vitro coculture is not tissue specific nor dependent on cell type.

The influence of oocyte maturity and embryo quality on pregnancy rate in a program for in vitro fertilization-embryo transfer.

An important factor influencing the pregnancy rate after in vitro fertilization-embryo transfer (IVF-ET) appears to be the number of embryos transferred to the uterus. In this study, the influence of oocyte maturity and embryo quality on pregnancy rate was assessed in patients undergoing IVF-ET. Ovarian hyperstimulation was performed by human menopausal gonadotropin (hMG [n = 29]), clomiphene citrate (CC)/hMG (n = 81), and hMG/follicle-stimulating hormone (FSH [n = 13]) protocols. Oocyte maturity was graded on a scale from 1 to 5 based on the morphology of the ooplasm, cumulus mass, corona radiata, and membrana granulosa cells. Embryos were graded according to the symmetry of the blastomeres and the presence or absence of fragmentation. Mature preovulatory oocytes yielded the highest fertilization rates. No differences were found among the protocols in terms of fertilization rate, embryo quality, or pregnancy rate. When all protocols were combined, patients who conceived had a significantly higher number of embryos transferred than those who did not conceive (3.6 +/- 0.1 [mean = SEM] versus 2.7 +/- 0.1). When embryo quality was compared, there was no difference in the number of "B" embryos transferred between patients who conceived and those who did not (1.2 +/- 0.2 versus 1.2 +/- 0.1), but the patients who conceived had significantly more "A" embryos transferred (1.6 +/- 0.3 versus 0.8 +/- 0.1). These data suggest that the treatment protocol did not determine embryo quality. Furthermore, the increase in pregnancy rates seen with an increase in embryos transferred is the result of the transfer of more "A" embryos.

Correlación entre las buenas prácticas de manufactura y el cumplimiento de los criterios microbiológicos en la fabricación de helados en Chile / Correlation between the good manufacturing practices and compliance with microbiological criteria in the manufacture of ice cream in Chile

The Good Manufacturing Practices (GMP) is a basic tool to obtain safe foodstuffs for human consumption. The main goal of this research was to determine the percentage of compliance with GMP in ice cream factories and its correlation with the acceptance of microbiological criteria in samples of ice cream, according to the Sanitary Chilean Food Regulation. Inspectors from the Ministerial Regional Secretariat (SEREMI) of Health Ñuble audited GMP in 40 ice cream factories and 435 microbial analyses from ice cream samples taken between the years 2005 and 2010 were analyzed. Results showed that 55.2% of samples failed in RAM (105 UFC/g) and coliform (102 UFC/g) and 4.6% for Staphylococcus aureus (102 UFC/g) and 100% for Salmonella spp. The biggest RAM was during summer with 60% rejection. Sixty seven% of inspections of GMP reached values above the minimum (70% compliance). Facilities were the best item assessed (88.4%), and training was the worst (20.3%). The correlation between the results of audits of GMP and microbiological criteria allowed proving that with a percentage equal to or greater than 80% compliance with GMP would ensure the microbiological quality of ice cream, thereby decreasing the risk for consumer health.

Las Buenas Prácticas de Manufacturas (BPM) son una herramienta básica para obtener un producto alimenticio seguro para consumo humano. El objetivo de este estudio fue determinar el porcentaje de cumplimiento de BPM y su correlación con la aceptación de criterios microbiológicos en fábricas y muestras de helados de Ñuble, Chile, de acuerdo al Reglamento Sanitario de Alimentos chilenos. Se analizaron auditorías de BPM aplicadas por fiscalizadores de Secretaría Regional Ministerial (SEREMI) de Salud Ñuble, a 40 fábricas de helados y los resultados microbiológicos de 435 muestras de helados entre los años 2005 y 2010. Se determinó que el 55,2% de las muestras no cumplieron para RAM (105 UFC/g) y coliformes (102 UFC/g) y 4,6% para Staphylococcus aureus (102 UFC/g), en cambio el 100% cumplieron para Salmonella spp. Los mayores RAM fueron en verano con 60% de rechazo. El 67% de auditorías de BPM alcanzaron valores superiores al mínimo (70% de cumplimiento), las instalaciones fue el ítem mejor evaluado (88,4%), y capacitación del personal el peor (20,3 %). La correlación entre los resultados de auditorías de BPM y criterios microbiológicos permitió comprobar que con un porcentaje igual o superior al 80% de cumplimiento de BPM se aseguraría la calidad microbiológica de helados, disminuyendo con ello el riesgo en la salud del consumidor.

[The influence of -238 and -308 TNF alpha polymorphisms on the pathogenesis and response to treatment in rheumatoid arthritis]. / Papel de los polimorfismos -238 y -308 del promotor del factor de necrosis tumoral alfa en la patogenia y respuesta al tratamiento anti-factor de necrosis tumoral alfa en artritis reumatoide.

Rheumatoid arthritis (RA) is a systemic autoimmune disease that affects 0.8% of the world population, it affects the synovial membrane of joints and the clinical presentation encompasses a wide spectrum, ranging from a mild to a severe and erosive disease that causes joint and cartilage destruction which finally provokes irreversible structural damage and patient disability. In the last years, there have been important advances in the pathogenesis of this disease, the efforts have been concentrated on pro-inflammatory cytokines such as tumor necrosis factor alpha (TNFalpha). This protein guides numerous events in the synovial and systemic inflammatory process and is encoded in the Major Histocompatibility Complex (MHC), one of the most polymorphic of the genome. Polymorphisms affecting the TNFalpha gene and its regulatory regions are associated with RA prevalence and course. There is a possible association between these polymorphisms and the clinical response to the use of monoclonal antibodies anti-TNFalpha. The possibility that the determination of genotypes -238 and -308 may have prognostic and therapeutic consequences is debated nowadays.

Assessment of human sperm functional changes after in-vitro coincubation with cells retrieved from the human female reproductive tract.

Human spermatozoa must undergo functional changes prior to fertilization; however, the site of this physiological event is still unclear. To evaluate the influence of the female reproductive tract on sperm fertilizing capacity, fertile sperm samples were coincubated with endometrial, oviductal, granulosa and cumulus cells, follicular fluid and maternal serum. Sperm penetration into the zona-free hamster ova and motion parameters were measured daily for 72 h. Compared to control samples, endometrial and oviductal cell cultures did not alter sperm fertilizing capacity or their movement characteristics. Sperm coincubated with follicular fluid, granulosa or cumulus cells exhibited a significantly higher ability to penetrate zona-free hamster ova for up to 48 h. Sperm motility increased at 4 h in the presence of follicular fluid and serum. At 24 h sperm velocity and amplitude of lateral head displacement significantly declined in sperm samples exposed to serum, and velocity also declined in follicular fluid and with coincubation using ovarian follicle cells. Sperm motility and velocity decreased at 48 h in the presence of serum, follicular fluid, cumulus or granulosa cells. Our findings may suggest that specific secretory factors produced in the human pre-ovulatory ovarian follicle enhance human sperm fertilizing capacity.