Altered Vaginal Microbiota Composition Correlates With Human Papillomavirus and Mucosal Immune Responses in Women With Symptomatic Cervical Ectopy.

Cervical ectopy is a benign condition of the lower genital tract that is frequently detected in women of reproductive age. Although cervical ectopy is regarded as a physiological condition, some women experience symptoms such as leucorrhoea, persistent bleeding and recurrent vaginal infections that require medical intervention. Cervical ectopy has not been linked to cervical cancer, but it is thought to facilitate the acquisition of sexually transmitted diseases (STDs), like Human Papillomavirus (HPV) infection, as it provides a favorable microenvironment for virus infection and dissemination. We and others have described the presence of oncogenic HPV types in women with symptomatic cervical ectopy. The relevance of this finding and the impact of symptomatic cervical ectopy on the cervicovaginal microenvironment (vaginal microbiota, immune and inflammatory responses) are currently unknown. To shed some light into the interplay between HPV, the vaginal microbiota and mucosal immune and inflammatory responses in the context of this condition, we enrolled 156 women with symptomatic cervical ectopy and determined the presence of HPV using a type-specific multiplex genotyping assay. Overall, HPV was detected in 54.48% women, oncogenic HPV types were found in more than 90% of HPV-positive cases. The most prevalent HPV types were HPV16 (29.4%), HPV31 (21.17%) and HPV18 (15.29%). Next, we evaluated the vaginal microbial composition and diversity by 16S rDNA sequencing, and quantified levels of cytokines and chemokines by flow cytometry using bead-based multiplex assays in a sub-cohort of 63 women. IL-21 and CXCL9 were significantly upregulated in HPV-positive women (p=0.0002 and p=0.013, respectively). Women with symptomatic cervical ectopy and HPV infection had increased diversity (p<0.001), and their vaginal microbiota was enriched in bacterial vaginosis-associated anaerobes (Sneathia, Shuttleworthia, Prevotella, and Atopobium) and depleted in Lactobacillus spp. Furthermore, the vaginal microbiota of women with symptomatic cervical ectopy and HPV infection correlated with vaginal inflammation (IL-1ß, rho=0.56, p=0.0004) and increased mucosal homeostatic response (IL-22, rho=0.60, p=0.0001). Taken together, our results suggest that HPV infection and dysbiotic vaginal communities could favor a vaginal microenvironment that might delay the recovery of the cervical epithelium in women with symptomatic cervical ectopy and favor STDs acquisition.

Management of calves in commercial dairy farms in Mecklenburg-Western Pomerania, Germany and its impact on calf mortality and prevalence of rotavirus and Cryptosporidium parvum infections in pre-weaned calves.

In a cross-sectional study, impact of management in dairy farms on calf mortality rates and prevalence of rotavirus and Cryptosporidium parvum in feces of calves was investigated. Sixty-two commercial dairy herds in Mecklenburg-Western Pomerania, Germany, were stratified selected in 2019. We performed in-person interviews and fecal specimens in samples of all-female calves of age 7 up to 21 days. Management data were documented on farm level. A Multiscreen Ag-ELISA was performed to determine rotavirus and Cryptosporidium parvum. Associations between two calf mortality rates, detection of C. parvum and rotavirus, and predictors were examined with GLM models. In farms with routine vaccination against respiratory diseases, 31-days mortality rate was 4.2% +/-1.26 compared to 7.6% +/-0.97 (p = 0.040) on non-vaccinating farms. Six-months mortality was lower in farms that continued feeding milk to calves during periods of diarrhea compared to farms that did not (6.9% +/-0.8 vs. 12.4% +/-2.3). In case of a routine shifting of calves from the calving box into calf boxes less C. parvum was detected compared to an individual moving of calves (33.3% +/-2.6 vs. 19.6% +/-5.3; p = 0.024). Our model confirms a positive association between occurrence of aqueous feces and frequency of detection of C. parvum (45.4% +/-23.6 vs. 21.4% +/-18.7; p < 0.001). Frequency of detection of rotavirus was lower in farms that reported a defined amount of applicated colostrum per calf than in farms that presented a range of colostrum instead of a defined amount. This study indicates the potential for mitigation of risk factors for mortality in calves.

Resultados funcionales e imagenológicos de la reparación artroscópica de lesiones parciales bursales del manguito rotador sin acromioplastia. Seguimiento a mediano plazo / Midterm functional outcomes and tendon integrity after in situ repair of partial bursal supraspinatus tears without acromioplasty

Antecedentes y objetivoAnalizar la cicatrización mediante ecografía y el seguimiento funcional a mediano plazo de pacientes con roturas parciales bursales del manguito rotador (RPBMR) con reparación in situ sin acromioplastia.Materiales y métodosSe evaluaron 62 pacientes. Se realizó la medición del rango de movimiento y la puntuación en el score American Shoulder and Elbow Surgeons (ASES). El dolor se registró usando una escala visual análoga (EVA). Se valoró ecográficamente la reparación con un mínimo de seguimiento de 5 años.ResultadosLa edad promedio fue de 57,2 años (rango de 44 a 77 años) y el seguimiento promedio, de 7 años (rango de 5 a 9 años). Todos los parámetros del rango de movimiento activo mejoraron significativamente (p<0,0001). El puntaje de ASES mejoró de 46,5 a 90,2 y los puntajes EVA mejoraron de 6,5 a 1,73 (p<0,0001). Un total de 56 pacientes (90%) realizaron control ecográfico al final del seguimiento; 51 presentaron integridad del tendón (91%) en la ecografía. Cinco pacientes presentaron re-roturas (9%); 3 tuvieron una re-rotura completa del tendón (5%) y 2 una re-rotura parcial (4%). No hubieron diferencias funcionales o del rango de movilidad significativas entre aquellos pacientes con el tendón intacto y los que presentaron re- roturas.ConclusionesA mediano plazo, la reparación in situ artroscópica de RPBMR demostró excelentes resultados funcionales en la mayoría de los pacientes con baja tasa de complicaciones. Estos mantienen un alto índice de cicatrización en el tiempo.(AU)

Background and purposeThe purpose of this study was to analyze midterm functional outcomes and tendon integrity in patients treated with in situ arthroscopic repair of Partial Bursal Rotator Cuff Tears (PBRCTs) without acromioplasty.Materials and methodsSixty two patients were included. Clinical assessment consisted of glenohumeral range of motion measurement and the American Shoulder and Elbow Surgeons (ASES) score. Pain was rated by using a visual analog scale (VAS). A postoperative ultrasound image control was performed at a minimum 5 years follow up to assess tendon integrity.ResultsMean age was 57.2 years (range, 44–77 years) and mean follow up was 7 years (range of 5–9 years). Significant improvements in range of motion, functional outcomes and pain were observed postoperatively (P<.0001). The ASES score improved from 46.5 to 90.2; and the VAS improved from 6.5 to 1.73 (P<.0001). Fifty six patients (90%) performed ultrasound evaluation at the end of follow up. Fifty-one patients (91%) presented tendon integrity on ultrasound. Five patients presented re-tears (9%). Three patients (5%) had a complete tendon re-tear and two patients (4%) had a partial re-tear. No difference in range of motion or functional outcomes were found between patients with intact tendon and those who had re-tears.ConclusionsIn the midterm follow up, arthroscopic in situ repair PBRCTs without acromioplasty showed excellent functional outcomes and high healing rates in most patients with low complication rates.(AU)

Midterm functional outcomes and tendon integrity after in situ repair of partial bursal supraspinatus tears without acromioplasty / Resultados funcionales e imagenológicos de la reparación artroscópica de lesiones parciales bursales del manguito rotador sin acromioplastia. Seguimiento a mediano plazo

Background and purposeThe purpose of this study was to analyze midterm functional outcomes and tendon integrity in patients treated with in situ arthroscopic repair of Partial Bursal Rotator Cuff Tears (PBRCTs) without acromioplasty.Materials and methodsSixty two patients were included. Clinical assessment consisted of glenohumeral range of motion measurement and the American Shoulder and Elbow Surgeons (ASES) score. Pain was rated by using a visual analog scale (VAS). A postoperative ultrasound image control was performed at a minimum 5 years follow up to assess tendon integrity.ResultsMean age was 57.2 years (range, 44–77 years) and mean follow up was 7 years (range of 5–9 years). Significant improvements in range of motion, functional outcomes and pain were observed postoperatively (P<.0001). The ASES score improved from 46.5 to 90.2; and the VAS improved from 6.5 to 1.73 (P<.0001). Fifty six patients (90%) performed ultrasound evaluation at the end of follow up. Fifty-one patients (91%) presented tendon integrity on ultrasound. Five patients presented re-tears (9%). Three patients (5%) had a complete tendon re-tear and two patients (4%) had a partial re-tear. No difference in range of motion or functional outcomes were found between patients with intact tendon and those who had re-tears.ConclusionsIn the midterm follow up, arthroscopic in situ repair PBRCTs without acromioplasty showed excellent functional outcomes and high healing rates in most patients with low complication rates.(AU)

Antecedentes y objetivoAnalizar la cicatrización mediante ecografía y el seguimiento funcional a mediano plazo de pacientes con roturas parciales bursales del manguito rotador (RPBMR) con reparación in situ sin acromioplastia.Materiales y métodosSe evaluaron 62 pacientes. Se realizó la medición del rango de movimiento y la puntuación en el score American Shoulder and Elbow Surgeons (ASES). El dolor se registró usando una escala visual análoga (EVA). Se valoró ecográficamente la reparación con un mínimo de seguimiento de 5 años.ResultadosLa edad promedio fue de 57,2 años (rango de 44 a 77 años) y el seguimiento promedio, de 7 años (rango de 5 a 9 años). Todos los parámetros del rango de movimiento activo mejoraron significativamente (p<0,0001). El puntaje de ASES mejoró de 46,5 a 90,2 y los puntajes EVA mejoraron de 6,5 a 1,73 (p<0,0001). Un total de 56 pacientes (90%) realizaron control ecográfico al final del seguimiento; 51 presentaron integridad del tendón (91%) en la ecografía. Cinco pacientes presentaron re-roturas (9%); 3 tuvieron una re-rotura completa del tendón (5%) y 2 una re-rotura parcial (4%). No hubieron diferencias funcionales o del rango de movilidad significativas entre aquellos pacientes con el tendón intacto y los que presentaron re- roturas.ConclusionesA mediano plazo, la reparación in situ artroscópica de RPBMR demostró excelentes resultados funcionales en la mayoría de los pacientes con baja tasa de complicaciones. Estos mantienen un alto índice de cicatrización en el tiempo.(AU)

Midterm functional outcomes and tendon integrity after in situ repair of partial bursal supraspinatus tears without acromioplasty. / Resultados funcionales e imagenológicos de la reparación artroscópica de lesiones parciales bursales del manguito rotador sin acromioplastia. Seguimiento a mediano plazo.

BACKGROUND AND PURPOSE: The purpose of this study was to analyze midterm functional outcomes and tendon integrity in patients treated with in situ arthroscopic repair of Partial Bursal Rotator Cuff Tears (PBRCTs) without acromioplasty. MATERIALS AND METHODS: Sixty two patients were included. Clinical assessment consisted of glenohumeral range of motion measurement and the American Shoulder and Elbow Surgeons (ASES) score. Pain was rated by using a visual analog scale (VAS). A postoperative ultrasound image control was performed at a minimum 5 years follow up to assess tendon integrity. RESULTS: Mean age was 57.2 years (range, 44-77 years) and mean follow up was 7 years (range of 5-9 years). Significant improvements in range of motion, functional outcomes and pain were observed postoperatively (P<.0001). The ASES score improved from 46.5 to 90.2; and the VAS improved from 6.5 to 1.73 (P<.0001). Fifty six patients (90%) performed ultrasound evaluation at the end of follow up. Fifty-one patients (91%) presented tendon integrity on ultrasound. Five patients presented re-tears (9%). Three patients (5%) had a complete tendon re-tear and two patients (4%) had a partial re-tear. No difference in range of motion or functional outcomes were found between patients with intact tendon and those who had re-tears. CONCLUSIONS: In the midterm follow up, arthroscopic in situ repair PBRCTs without acromioplasty showed excellent functional outcomes and high healing rates in most patients with low complication rates.

Does whole body vibration exercise improve oxidative stress markers in women with fibromyalgia?

The objective of this study was to investigate the effect of whole body vibration (WBV) exercise on oxidative stress markers in a group of women with fibromyalgia (FM) compared to a group of healthy women (CT). Twenty-one women diagnosed with FM and 21 age- and weight-matched healthy women were enrolled the study. Plasma oxidative stress markers (primary outcomes) were evaluated at rest and after WBV, and included thiobarbituric acid reactive substances (TBARS), iron reduction capacity (FRAP), superoxide dismutase antioxidant enzymes activity (SOD), and catalase (CAT). At rest, the FM group had higher TBARS (P<0.001) and FRAP (P<0.001), and lower CAT (P=0.005) compared to the CT. In the CT group, the WBV had no effect on TBARS (P=0.559) and FRAP (P=0.926), whereas it increased both SOD (P<0.001) and CAT (P<0.001). In the FM group, the WBV reduced TBARS (p <0.001), FRAP (P<0.001), and CAT (P=0.005), while it increased SOD (P=0.019). There was an interaction effect (moments vs groups) in the TBARS (effect size=1.34), FRAP (effect size=0.93), CAT (effect size=1.45), and SOD (effect size=1.44) (P<0.001). A single trial of WBV exercise improved all oxidant and antioxidant parameters towards a greater adaptation to the stress response in FM women.

Does whole body vibration exercise improve oxidative stress markers in women with fibromyalgia?

The objective of this study was to investigate the effect of whole body vibration (WBV) exercise on oxidative stress markers in a group of women with fibromyalgia (FM) compared to a group of healthy women (CT). Twenty-one women diagnosed with FM and 21 age- and weight-matched healthy women were enrolled the study. Plasma oxidative stress markers (primary outcomes) were evaluated at rest and after WBV, and included thiobarbituric acid reactive substances (TBARS), iron reduction capacity (FRAP), superoxide dismutase antioxidant enzymes activity (SOD), and catalase (CAT). At rest, the FM group had higher TBARS (P<0.001) and FRAP (P<0.001), and lower CAT (P=0.005) compared to the CT. In the CT group, the WBV had no effect on TBARS (P=0.559) and FRAP (P=0.926), whereas it increased both SOD (P<0.001) and CAT (P<0.001). In the FM group, the WBV reduced TBARS (p <0.001), FRAP (P<0.001), and CAT (P=0.005), while it increased SOD (P=0.019). There was an interaction effect (moments vs groups) in the TBARS (effect size=1.34), FRAP (effect size=0.93), CAT (effect size=1.45), and SOD (effect size=1.44) (P<0.001). A single trial of WBV exercise improved all oxidant and antioxidant parameters towards a greater adaptation to the stress response in FM women.

Parental-reported neurodevelopmental issues in Loeys-Dietz syndrome.

BACKGROUND: Loeys-Dietz syndrome (LDS) is a congenital multisystem disorder affecting the cardiovascular and musculoskeletal system. Limited data have reported neurodevelopmental (ND) issues in LDS. AIMS: To determine the extent of ND issues in patients with LDS. METHODS: A prospective study was performed of LDS patients or their caregivers. The study included data collected via an online survey of age-specific questions. Standard statistical methods were used for baseline and demographic characteristics, as well as group comparisons. OUTCOMES: Data were obtained from 67 patients with LDS (54% female). Median age was 14.9 years. Gene mutations included TGFBR1 (39%), TGFBR2 (40%), SMAD3 (7%), and unknown (14%). Motor delays (30%, 18/61) and hypotonia (63%, 37/60) occurred frequently. Physical (62%, 39/62), occupational (41%, 23/56), and speech therapies (34%, 20/58) were common. Feeding issues were common (41%, 23/56). TGFBR1 mutations were more frequent among those with motor delays and feeding issues. CONCLUSIONS: Patients with LDS and/or their caregivers report at least one ND problem in most cases, and many require therapies. These data suggest ND disorders should be considered to be part of the phenotype.

Lactation Biology Symposium: role of colostrum and colostrum components on glucose metabolism in neonatal calves.

In neonatal calves, nutrient intake shifts from continuous glucose supply via the placenta to discontinuous colostrum and milk intake with lactose and fat as main energy sources. Calves are often born hypoglycemic and have to establish endogenous glucose production (eGP) and gluconeogenesis, because lactose intake by colostrum and milk does not meet glucose demands. Besides establishing a passive immunity, colostrum intake stimulates maturation and function of the neonatal gastrointestinal tract (GIT). Nutrients and nonnutritive factors, such as hormones and growth factors, which are present in high amounts in colostrum of first milking after parturition, affect intestinal growth and function and enhance the absorptive capacity of the GIT. Likely as a consequence of that, colostrum feeding improves the glucose status in neonatal calves by increasing glucose absorption, which results in elevated postprandial plasma glucose concentrations. Hepatic glycogen concentrations rise much greater when colostrum instead of a milk-based colostrum replacer (formula with same nutrient composition as colostrum but almost no biologically active substances, such as hormones and growth factors) is fed. In contrast, first-pass glucose uptake in the splanchnic tissue tended to be greater in calves fed formula. The greater plasma glucose rise and improved energy status in neonatal calves after colostrum intake lead to greater insulin secretion and accelerated stimulation of anabolic processes indicated by enhanced maturation of the postnatal somatotropic axis in neonatal calves. Hormones involved in stimulation of eGP, such as glucagon and cortisol, depend on neonatal diet, but their effects on eGP stimulation seem to be impaired. Although colostrum feeding affects systemic insulin, IGF-I, and leptin concentrations, evidence for systemic action of colostral insulin, IGF-I, and leptin in neonatal calves is weak. Studies so far indicate no absorption of insulin, IGF-I, and leptin from colostrum in neonatal calves, unlike in rodents where systemic effects of colostral leptin are demonstrated. Therefore, glucose availability in neonatal calves is promoted by perinatal maturation of eGP and colostrum intake. There may be long-lasting effects of an improved colostrum supply and glucose status on postnatal growth and development, and colostrum supply may contribute to neonatal programming of performance (milk and growth) in later life, but data proving this concept are missing.

Anemia in kidney transplants without erythropoietic agents: levels of erythropoietin and iron parameters.

AIM: To study the association between hemoglobin, endogenous erythropoietin (EPO) levels and ferric parameters in kidney recipients not treated with EPO-stimulating agents. MATERIALS AND METHODS: Transverse study of 219 kidney transplant outpatients. The median time after transplantation was 54 months (P(25-75), 23-107). We assessed blood counts, ferric parameters, EPO levels, renal function (MDRD-4), and adjuvant treatment. We performed a linear regression analysis to predict hemoglobin. RESULTS: Median EPO values were 14.05 mUI/mL (P(25-75) = 10.2-19.7). Applying the formulas described by Beguin, kidney transplant recipients showed a low observed/expected ratio of erythropoietin and of transferrin. Considering anemia to be an hemoglobin of < 12 g/dL in women and < 13 g/dL in men, 24.2% of subjects were anemic (n = 53), including 2.3% with hemoglobin < 11 g/dL. Anemic patients displayed worse renal function (49.2 ± 18.5 versus 55.46 ± 16.58 mL/min/1.73 m(2) in nonanemic; P = .021). There were no differences in C-reactive protein. The patients receiving a combination of angiotensin-converting enzyme inhibitors (ACEI) and angiotensin II receptor blockers (ARB) showed the highest prevalence of anemia compared with other groups (42.9%, P = .027). EPO levels were significantly lower among patients treated with these drugs (P = .041), without differences in transferrin and ferritin. The percentage of anemic patients treated with mammalian target of rapamycin inhibitors (mTORi) was 31% versus 22.2% among those not receiving these immunosuppressants (P = .23). Although there were no differences in hemoglobin levels, patients treated with mTORi, showed higher EPO levels (P = .005) and lower mean corpuscular volume (P < .001). Regarding the etiology of chronic kidney disease, less frequently anemic patients were those with polycystic kidney disease (8.6% versus 26.7% in the rest, P = .021). The formula obtained by multiple linear regression to calculate hemoglobin was: hemoglobin = 11829-0909 log (EPG level) - 0455 (if female) + 0.010 0.013 transferrin + 0.013 creatinine clearance (r = .424, P < .001). CONCLUSIONS: Treatment with ACEI and/or ARBs seemed to produce a defect in the synthesis of EPO, while those treated with mTORi, a hyporesponsive state.

New developments in understanding interfacial processes in turbulent flows.

Interfaces, across which fluid and flow properties change significantly, are a ubiquitous feature of most turbulent flows and are present within jets, plumes, homogeneous turbulence, oceans and planetary atmospheres. Even when the interfaces occupy a small volume fraction of the entire flow, they largely control processes such as entrainment and dissipation and can act as barriers to transport. This Theme Issue brings together some of the leading recent developments on interfaces in turbulence, drawing in many methodologies, such as experiments, direct number simulations, inverse methods and analytical modelling.

Distribution of Phoma sclerotioides on Alfalfa and Winter Wheat Crops in the North Central United States.

Brown root rot of alfalfa (Medicago sativa), caused by Phoma sclerotioides, has been reported in several states in the northern United States and in western Canada. A survey was conducted to determine the distribution of the fungus in Minnesota and Wisconsin. Isolates of the pathogen were recovered from roots of alfalfa, winter wheat, and perennial ryegrass plants. The internal transcribed spacer (ITS) 1, 5.8S, and ITS2 of the rDNA of the isolates from alfalfa and wheat were identical and matched the sequences of a P. sclerotioides isolate from Wyoming. The fungus was found to be widespread in both states and was detected in roots of alfalfa plants from 17 counties in Minnesota and 14 counties in Wisconsin using polymerase chain reaction (PCR)-based assays. A real-time PCR assay was developed that increased sensitivity of detecting the pathogen from plant tissues and soil. The isolates from alfalfa caused disease on inoculated winter wheat plants. Although the fungus was previously found associated with roots of diseased cereal and turfgrass plants, this is the first demonstration of pathogenicity of P. sclerotioides on wheat.

Reactogenicity and immunogenicity profile of a two-dose combined hepatitis A and B vaccine in 1-11-year-old children.

This study was conducted to compare the reactogenicity, immunogenicity and safety of a combined two-dose (0, 6 months) hepatitis A and B vaccine (720ELU HAV, 20 mcg HBsAg) with the established three-dose (0, 1 and 6 months) hepatitis A and B vaccine (360ELU HAV, 10 mcg HBsAg). A total of 511 children aged 1-11 years who had not previously received a hepatitis A or B vaccine were enrolled in the study. Both vaccines were well tolerated, and were shown to be safe and immunogenic. The analysis, stratified according to two age groups (1-5 year and 6-11-year-old children) demonstrated that the reactogenicity profile of the two-dose schedule was at least as good as that of the established schedule. Both vaccines and schedules provided at least 98% seroprotection against hepatitis B and 100% seroconversion against hepatitis A, 1 month after the end of the vaccination course (Month 7).

Comparison of the reactogenicity and immunogenicity of a combined diphtheria, tetanus, acellular pertussis, hepatitis B, inactivated polio (DTPa-HBV-IPV) vaccine, mixed with the Haemophilus influenzae type b (Hib) conjugate vaccine and administered as a single injection, with the DTPa-IPV/Hib and hepatitis B vaccines administered in two simultaneous injections to infants at 2, 4 and 6 months of age.

An open, randomised, multicentre trial was performed to compare the reactogenicity and safety profile of the administration of a hexavalent diphtheria-tetanus-acellular pertussis-hepatitis B-inactivated polio (DTPa-HBV-IPV) vaccine administered in one injection mixed with Haemophilus influenzae type b (Hib) conjugate vaccine (Group 1) with that of a pentavalent DTPa-IPV vaccine mixed with a Hib vaccine (DTPa-IPV/Hib), simultaneously administered with HBV (Group 2) in two injections in opposite thighs, as a primary vaccination course, to healthy infants at 2, 4 and 6 months of age. A total of 235 completed the study, 120 from Group 1 and 115 from Group 2. Blood samples (pre-vaccination and 1 month after the third dose) were obtained from a subset of infants (Group 1: 40; Group 2: 31) to assess the immune response to vaccination. Local and general solicited symptoms were recorded by parents on diary cards. Seven hundred and five diary cards (Group 1: 360; Group 2: 345) were collected. The clinically relevant and most commonly reported local reaction was pain (infant cried when the limb was moved) in 2.5% (Group 1) and 1.2% (Group 2) of diary cards. Fever was more frequently reported in Group 1 (21% of diary cards) than in Group 2 (12% of diary cards). However only 3 and 2% of doses in Groups 1 and 2, respectively, were responsible for a rectal temperature between 38.6 and 39.5 degrees C and only one case (Group 2) had > or =39.5 degrees C. Other clinically relevant general symptoms were rarely recorded: irritability (2-2.8%), loss of appetite (0.3-0.6%) and drowsiness (0.3-0.3%). All subjects included in the immunogenicity analysis had seroprotective titres to diphtheria, tetanus, polio virus types 1 and 3, Hib. Almost all subjects were seroprotected for anti-polio type 2 and hepatitis B (with the exception of 1 subject in Group 1 for each antigen). The vaccines response rates to pertussis antigens were over 97 and 90% in Groups 1 and 2, respectively. This study shows that, from a clinical perspective, the DTPa-HBV-IPV/Hib vaccine given in a single injection has a similar reactogenicity and safety profile to that of two licensed vaccines (DTPa-IPV/Hib, HBV) given in two simultaneous injections to infants at 2, 4 and 6 months of age. This is a valuable advantage, since in some countries, such as Spain and the UK, an additional injection (for the administration of meningococcal C conjugate vaccine) has been recently included in the infants' vaccination calendars.

Determination of the fraction of S-phase cells in root meristems using bromodeoxyuridine labeling.

The use of bromodeoxyuridine (BrdU) and subsequent immunocytochemical visualization for studying cell proliferation in plant meristems was investigated in Allium cepa L. root-tips. We describe the optimization of an indirect immunoperoxidase method for detecting incorporation of this DNA precursor in pulse-labeled cells. The basic object of this study is to quantify the extent to which the fraction of S-phase cells can reliably be estimated in asynchronous populations. A matrix of parallel labeling schedules with tritiated-thymidine or BrdU was developed, and the labeling indices provided by autoradiography or immunocytochemistry were compared. Thus, 0.5 mM BrdU assured saturation S-phase labeling after an exposure time of 30 min, and the mean length of the S-phase determined under such conditions was similar to that previously reported for this plant system. Interestingly, Feulgen staining did not interfere with subsequent detection of the BrdU probe. This allowed comparative evaluations of the nuclear DNA content by Feulgen-microdensitometry and the position of a given cell in G1, S or G2 compartments. We also explored the possibility of quantifying BrdU-incorporation in single nuclei by densitometry measurement of the peroxidase label.