Decline in Vaccine-Type Streptococcus pneumoniae Serotypes Following Pneumococcal Conjugate Vaccine Introduction in Madagascar.

BACKGROUND: The 10-valent conjugate vaccine (PCV10) was introduced into the Extended Program on Immunization in Madagascar. We assessed the impact of PCV10 on the targeted pneumococcal serotypes among children < 5 years of age at Centre Hospitalier Universitaire Mère Enfant Tsaralalàna. METHOD: Between 2012 and December 2018, cerebrospinal fluid (CSF) samples were collected and tested for S. pneumoniae by culture, and antigen tests. The Sentinel Site Laboratory (SSL) referred available CSF samples to the Regional Reference Laboratory (RRL) for real-time polymerase chain reaction confirmatory testing and serotyping. RESULTS: In total, 3616 CSF specimens were collected. The SSL referred 2716 to the RRL; 125 were positive for S. pneumoniae. At the RRL, 115 samples that tested positive for S. pneumoniae were serotyped; PCV10 serotypes accounted for 20%. Compared to the pre-PCV period, the proportion of S. pneumoniae detected declined from 22% to 6.6%, (P < .05), the proportion of PCV10 serotypes as the cause of pneumococcal meningitis cases declined by 26% following vaccine introduction. CONCLUSIONS: In our findings, PCV10 introduction resulted in a decline of meningitis caused by S. pneumoniae and PCV10 vaccine serotypes.

Characterization of Gd loaded chitosan-TPP nanohydrogels by a multi-technique approach combining dynamic light scattering (DLS), asymetrical flow-field-flow-fractionation (AF4) and atomic force microscopy (AFM) and design of positive contrast agents for molecular resonance imaging (MRI).

Chitosan CS-tripolyphosphate TPP/hyaluronic acid HA nanohydrogels loaded with gadolinium chelates (GdDOTA ⊂ CS-TPP/HA NGs) synthesized by ionic gelation were designed for lymph node (LN) MRI. In order to be efficiently drained to LNs, nanogels (NGs) needed to exhibit a diameter ϕ < 100 nm. For that, formulation parameters were tuned, using (i) CS of two different molecular weights (51 and 37 kDa) and (ii) variable CS/TPP ratio (2 < CS/TPP < 8). Characterization of NG size distribution by dynamic light scattering (DLS) and asymetrical flow-field-flow-fractionation (AF4) showed discrepancies since DLS diameters were consistently above 200 nm while AF4 showed individual nano-objects with ϕ < 100 nm. Such a difference could be correlated to the presence of aggregates inherent to ionic gelation. This point was clarified by atomic force microscopy (AFM) in liquid mode which highlighted the main presence of individual nano-objects in nanosuspensions. Thus, combination of DLS, AF4 and AFM provided a more precise characterization of GdDOTA ⊂ CS-TPP/HA nanohydrogels which, in turn, allowed to select formulations leading to NGs of suitable mean sizes showing good MRI efficiency and negligible toxicity.

A new magnetic resonance imaging contrast agent loaded into poly(lacide-co-glycolide) nanoparticles for long-term detection of tumors.

The incorporation of a lipophilic Gd chelate (GdDO3A-C12) in biocompatible PLGA poly(D, L-lactide-co-glycolide) nanoparticles was explored as an approach to increase the relaxivity of contrast agents for magnetic resonance imaging. By nanoprecipitation, it was possible to obtain PEGylated gadolinium nanoparticles (mean diameter of 155 nm) with high Gd loading (1.1 × 10(4) Gd centers per nanoparticle). The corresponding GdDO3AC12 ⊂ NPs nanoparticles exhibited an enhanced relaxivity (up to sixfold greater than DOTAREM® at 40 MHz) because the nanoparticle framework constrained the lipophilic Gd chelate motion and favorably impacted the Gd chelate rotational correlation time. T1-weighted imaging at 3 T on phantoms showed enhanced contrast for the GdDO3AC12 ⊂ NPs. Importantly, Gd chelate leakage was almost nonexistent, which suggested that these GdDO3AC12 ⊂ NPs could be useful for long-term MRI detection.

Vectorization of copper complexes via biocompatible and biodegradable PLGA nanoparticles.

A double emulsion-solvent diffusion approach with fully biocompatible materials was used to encapsulate copper complexes within biodegradable nanoparticles, for which the release kinetics profiles have highlighted their potential use for a prolonged circulating administration.

Evaluation of mTHPC-loaded PLGA nanoparticles for in vitro photodynamic therapy on C6 glioma cell line.

Photodynamic therapy (PDT) is a very attractive strategy to complement or replace common cancer treatments such as radiotherapy, surgery, and chemotherapy. Some molecules have shown their efficiency as photosensitizers (PS), still many issues have to be solved such as the inherent cytotoxicity of the PS or its hydrophobic properties causing limitation in their solubility, leading to side effects. In this study, the encapsulation of an approved PS, the meso-tetra hydroxyphenylchlorine (mTHPC, Foscan®) within biocompatible and biodegradable poly(D, l-lactide-co-glycolide) acid (PLGA) NPs prepared by the nanoprecipitation method was studied. The mTHPC-loaded NPs (mTHPC ⊂ PLGA NPs) were analyzed by UV-vis spectroscopy to determine the efficiency of mTHPC encapsulation, and by dynamic light scattering (DLS) and atomic force microscopy (AFM) to determine mTHPC ⊂ PLGA NPs sizes, morphologies and surface charges. The longitudinal follow-up of mTHPC release from the NPs indicated that 50% of the encapsulated PS was retained within the NP matrix after a period of five days. Finally, the cytotoxicity and the phototoxicity of the mTHPC ⊂ PLGA NPs were determined in murine C6 glioma cell lines and compared to the ones of mTHPC alone. The studies showed a strong decrease of mTHPC cytotoxicity and an increase of mTHPC photo-cytotoxicity when mTHPC was encapsulated. In order to have a better insight of the underlying cellular mechanisms that governed cell death after mTHPC ⊂ PLGA NPs incubation and irradiation, annexin V staining tests were performed. The results indicated that apoptosis was the main cell death mechanism.

Trichomonas vaginalis among women raped in Antananarivo, 2005-2011.

Rape is a forensic emergency due to the possibility of sexually transmitted infections, including HIV/AIDS. Moreover, an upsurge is currently underway. The objective of this study was to determine the incidence of Trichomonas vaginalis in women undergoing tests at the Microbiology Unit of the Joseph Ravoahangy Andrianavalona Antananarivo Hospital after being raped. This retrospective descriptive study covers the 7-year period from 2005 to 2011 and reports the results of direct examination of the vaginal discharge after vulvar smears from rape patients. We studied the age of patients and the results of these direct smear examinations that tested for T. vaginalis. In our study, the number of women with T. vaginalis increased markedly with age. The mean age of the patients was 13.7 years (range: 2 to 41 years). Among the 232 rape victims tested, only 26 were positive for it (11.20%); trichomoniasis was thus rare among these women. T. vaginalis is a common cause of vaginitis in women and one of the most common infections transmitted by sexual assault. In children, the presence of T. vaginalis in vaginal sample is a strong indicator of sexual abuse. Other sexually transmitted infections can be transmitted during sexual abuse.

Hydroxyethylstarch microcapsules: a preliminary study for tumor immunotherapy application.

The objective of this work was to prepare microcapsules which would allow protection and slow release of antigens used for melanoma immunotherapy treatment. Hydroxyethylstarch (HES) microcapsules were prepared using interfacial cross-linking with terephthaloyl chloride (TC). They were characterized with respect to morphology (microscopy) and size (in the 4-15 microm range). Bovine serum albumin (BSA) was used as model protein for loading and release studies. Microcapsules were loaded with solutions at different protein concentrations (0.5-5%). The maximum loading efficiency (20%) was observed with the concentration of 2.5%, which allowed a loading capacity near 100%. Confocal laser scanning microscopy (CLSM) visualization showed that BSA was entrapped within the microcapsules and not only associated to their outer surface. BSA-release studies showed a 20% BSA release within 30 min while 80% remained entrapped in the microcapsules for 4 days. Microcapsules were degraded by alpha-amylase and addition of esterase to alpha-amylase enhanced slightly their degradation. In vitro studies on melanoma cells showed that HES microcapsules were non-toxic. Preliminary in vivo studies demonstrated that microcapsules were biodegradable after intraperitoneal injection (i.p.). The observation of peritoneal wash showed a complete degradation within 7 days, indicating a possible application as an in vivo drug delivery system especially to enhance the presentation of antigens.

Sunscreen use, wearing clothes, and number of nevi in 6- to 7-year-old European children. European Organization for Research and Treatment of Cancer Melanoma Cooperative Group.

BACKGROUND: Previous epidemiologic studies have suggested that sunscreen use is associated with an increased risk of melanoma skin cancer. Because high nevi (mole) count in adults is a strong predictor of melanoma, we conducted a study examining the number of nevi in 6- to 7-year-old European children, according to their sunscreen use. METHODS: Whole-body and site-specific counts of nevi 2 mm or larger were performed in 631 children in their first year of primary school in four European cities. Independently, parents were interviewed regarding sun exposure, sunscreen use, and physical sun protection of their child. RESULTS: After adjustment for sun exposure and host characteristics (e.g., skin phototype, eye color), the relative risk for high nevus count on the trunk was 1.68 (95% confidence interval [CI] = 1.09-2.59) for the highest level of sunscreen use and 0.59 (95% CI = 0.36-0.97) for the highest level of wearing of clothes while in the sun. The sun protection factor had no effect on nevus counts despite a high median value of 17.4. Sunburn number was not associated with nevus count. The highest risk associated with sunscreen use was found among children who had never experienced sunburn. CONCLUSIONS: In white, European children, sunscreen use appears to be associated with development of nevi, probably because it allows longer sun exposures. Wearing clothes may be an effective way to prevent proliferation of nevi. Since a high nevus count is a strong predictor of melanoma, sunscreen use may be involved in melanoma occurrence because it may encourage recreational sun exposure.

Visual activity in the telencephalon of the painted turtle, Chrysemys picta.

Multiple unit activity in response to visual stimulation was recorded in the following telencephalic areas of the painted turtle: lateral and medial divisions of the dorsal cortex, lateral and medial divisions of the dorsal ventricular ridge, and the striatum. The data confirm anatomical evidence for ascending visual input to the dorsal cortex, the lateral dorsal ventricular ridge and the striatum. The identification of a new visual zone in a medial division of the dorsal ventricular ridge suggests that turtles, and perhaps other reptiles, possess at least 3 visual pallial areas.

Fourier transform infrared spectroscopic studies of cross-linked human serum albumin microcapsules. 3. Influence of terephthaloyl chloride concentration on spectra and correlation with microcapsule morphology and size.

Microcapsules were prepared from human serum albumin (HSA) by interfacial cross-linking with terephthaloyl chloride (TC). TC concentrations were increased from 0.5 to 5% w/v, while pH (9.8) and reaction time (30 min) were kept constant. Fourier transform infrared (FT-IR) spectra of lyophilized microcapsules were compared. Correlations were established with microcapsule morphology and size. The results were compared with those of previous studies exploring pH or reaction time and with those of parallel determinations of microcapsule free amino groups. With 0.5% TC, decreases of the ester-assigned 1724-cm-1 band area and of the carboxylate-assigned 1394-cm-1 band area were observed compared with pure HSA. This phenomenon was attributed to a removal of contaminating lipids of HSA. Increasing TC concentration resulted in a progressive increase of the band areas at 1724 cm-1 (esters) and 1795 cm-1 (anhydrides), in a further decrease of the 1394-cm-1 band area (carboxylates), and in marked alterations of teh 1340-1080-cm-1 region. These changes, which revealed the progressive acylation of hydroxy and carboxylate groups of HSA, were accompanied by an increase of the 1624-cm-1 band area (beta-sheet), reflecting interchain H-bonding due to cross-linking. As observed in the previous studies of pH and reaction time, important spectral changes corresponded to low values of -NH2 content, to a decrease in microcapsule mean size (from > 30 to < 15 microns), and to modifications of the membrane surface (made rough).

Fourier transform infrared spectroscopic studies of human serum albumin microcapsules prepared by interfacial cross-linking with terephthaloylchloride: influence of polycondensation pH on spectra and relation with microcapsule morphology and size.

Fourier transform infrared (FT-IR) spectroscopic studies were performed on microcapsules prepared through interfacial cross-linking of human serum albumin (HSA) with terephthaloylchloride at various pH values (5.9 to 11). Correlations were established with morphology and size of microcapsules. Increasing polycondensation pH resulted notably in a progressive increase of peaks at 1795 and 1724 cm-1, assigned to anhydride and ester; respectively, in a decrease of the carboxylate-assigned 1394 cm-1 peak, and in alterations of the 1340-1080-cm-1 region. These spectral changes were most pronounced from pH 9 and were shown to correspond to smaller-sized microcapsules (mean size decreased from 30-40 microns to less than 15 microns) with rough surfaces. Further soaking of highly cross-linked microcapsules in a pH 7.5 buffer resulted in the disappearance of the 1795 cm-1 peak, with a concurrent increase of the 1394 cm-1 peak and a decrease of the 1724 cm-1 peak. These changes, attributed to complete breaking of anhydride and partial hydrolysis of esters, were accompanied by an unwrinkling of the microcapsule membrane, then made smooth, and a significant increase in size. Treating microcapsules with hydroxylamine under alkaline conditions allowed complete reversal of the spectral alterations assigned to anhydride and ester formation. A comparable (slightly higher) increase in size was observed with microcapsules which exhibited smooth surfaces and a low density.

Fourier-transform infrared spectroscopic studies of cross-linked human serum albumin microcapsules. 2. Influence of reaction time on spectra and correlation with microcapsule morphology and size.

Microcapsules were prepared from human serum albumin (HSA) through interfacial cross-linking with terephthaloyl chloride (TC). Reaction times were increased from 2 to 60 min, while pH (9.8) and TC concentration (2.5% w/v) were kept constant. Fourier-transform infrared (FT-IR) spectra of lyophilized microcapsules were compared. Correlations were established with microcapsule morphology and size, as had been done in a previous study exploring the effect of increasing pH values. Microcapsules obtained after 2 min had to be considered separately. Minor alterations were observed in the spectrum as compared with pure HSA. They consisted of a decrease of the ester-assigned 1724-cm-1 band and of the carboxylate-assigned 1394-cm-1 band, attributed to a removal of contaminating lipids of HSA, and an increase of the 1624-cm-1 band, attributed to interchain H bonding following acylation of the NH2 groups. Prolonging the reaction time resulted in a progressive increase of the bands at 1724 (esters), 1795 (anhydrides), and 1624 cm-1 (beta-sheet), in a further decrease of the 1394-cm-1 band (carboxylates), and in marked alterations of the 1340-1080-cm-1 region. These important changes, which appeared after 5 min, reflect the progressive acylation of the hydroxy and carboxylate groups of HSA. As in the previous series of pH-based assays, important spectral changes were shown to correspond to a decrease in microcapsule mean size (from 32 to < 15 microns) and in important modifications of the membrane surface, made rough.

Cross-linked beta-cyclodextrin microcapsules. II. Retarding effect on drug release through semi-permeable membranes.

Microcapsules were prepared by interfacial cross-linking of beta-cyclodextrins (beta-CD) with terephthaloylchloride (TC) as described previously. Complexation assays were conducted with propranolol HCl. After 1 h incubation of 50 mg lyophilized microcapsules in 10 ml propranolol solution, the amounts of fixed drug were 507.5+/-8.6 micromol and 811.2+/-16.0 micromol per g lyophilized microcapsules with 1 mM and 2 mM solutions, respectively. A dialysis experiment was then performed. After 1 h incubation of microcapsules (10 or 50 mg) in 10 ml of 2 mM propranolol solution, the suspension was dialysed against a phosphate buffer pH 7.4 at 37 degrees C. The drug diffusion was all the more retarded that the amount of added beta-CD microcapsules was higher. Finally, double microcapsules were prepared using a suspension of beta-CD microcapsules (10-100 mg) in a solution of methylene blue in an acetate buffer pH 7.4. After adding human serum albumin (HSA), the suspension was emulsified in cyclohexane and double microcapsules were obtained by cross-linking the HSA with TC. In vitro release studies showed that the incorporation of beta-CD microcapsules resulted in a decrease in release rate of methylene blue, the decrease being related to the amount of encapsulated beta-CD microcapsules. The study then suggests interesting applications of beta-CD microcapsules for modulating the release rate of drugs through semi-permeable membranes.

Cross-linked beta-cyclodextrin microcapsules: preparation and properties.

Microcapsules were prepared by interfacial cross-linking of beta-cyclodextrins (beta-CD) with terephthaloyl chloride (TC). Batches were prepared from beta-CD solutions in 1 M NaOH, using 5% TC and a 30 min reaction time. Microcapsules were studied with respect to morphology (microscopy), size (laser diffraction technique) and, for selected batches, IR spectroscopy, determination of beta-CD content (polarimetry after alkaline dissolution of microcapsules) and complexing properties, evaluated using p-nitrophenol (pNP) as the guest molecule. Well-formed microcapsules were obtained from 5, 7.5, and 10% beta-CD solutions. The mean size of all batches was in the 10-35 microm range. The IR spectrum showed bands at 1724, 1280 and 731 cm(-1), reflecting the formation of esters. The beta-CD contents were 46, 56-58 or 60-66% for batches prepared from 5, 7.5 or 10% beta-CD solutions, respectively. The experiments conducted with 1 mM pNP showed a rapid complexation reaching a maximum within 1 h. When incubating 50 mg lyophilized microcapsules in 10 ml pNP solution, the maximal fixation (97.8 micromol/g microcapsules) was observed for small-sized particles ( approximately 11 microm) prepared from a 7.5% beta-CD solution. The method then appears as a simple and rapid procedure to provide stable microcapsules, having an interesting guest-binding ability.

Comparison of biochemical and immunoenzymatic macromethods and a new immunocytochemical micromethod for assaying estrogen receptors in human breast carcinomas.

Estrogen receptors (ERs) were assayed in 23 breast carcinomas by: (1) the conventional biochemical assay with dextran-coated charcoal (DCC); (2) the immunoenzymatic assay using a monoclonal antibody (MAb), ER-EIA (Abbott); and (3) an original cytochemical method using another MAb, ER-ICA (Abbott). The first two techniques were performed on biopsy samples, whereas the last was carried out on fine needle aspiration (FNA) samples. The ER contents in aspirates were evaluated by: (1) scaled proportions of colored neoplastic cells; (2) scaled coloration intensity; (3) total grading (= proportion plus intensity); (4) product grading (= proportion times intensity); and (5) a new index (NI) described in this paper. The ER-EIA assay correlated best, with a high statistical significance, with the NI (P less than .001); NI was also the only index that significantly correlated (P less than .05) with the DCC results. The results show that the ER-ICA assay offers the great advantages of being applicable to FNA specimens and of producing rapidly available results. This new technique enriches the panel of MAbs for the diagnosis of adenocarcinomas and offers a new tool for the therapeutic follow-up of breast cancer patients. Our preliminary results suggest that the anti-ER MAbs might be helpful for measuring the hormone dependence of small lesions not assayable by DCC, even under endocrine therapy, thus avoiding false-negative assays.

[Microcapsules with modulable properties made with cross-linked proteins and polysaccharides]. / Microcapsules à propriétés modulables formées de protéines et de polysaccharides coréticulés.

A cross-linking process was applied to mixtures of a protein (gelatin A or B) and a polysaccharide. Mixed-walled microcapsules were then obtained, which exhibited properties different from those of microcapsules prepared from the protein alone. They were shown to be more resistant to enzymatic lysis and this effect depended on the ratio of the polysaccharide, on its nature and on the cross-linking pH. Very hydrophilic microcapsules were obtained through cross-linking of gelatin admixed with alginate or carboxymethylcellulose. The addition of alginate to gelatin resulted in a slower release of encapsulated pilocarpine.

[Belgian registry of thyroid cancer. Preliminary epidemiological characteristics revealed by a retrospective study (1988-1995). Belgian Thyroid Cancer Study Group]. / Le registre belge du cancer thyroïdien. Premières caractéristiques épidémiologiques révélées par une étude rétrospective (1988-1995). Belgian Thyroid Cancer Study Group.

The aim of a cancer registry is to study the incidence of cancer in a well-determined population and to allow epidemiological research to the setting up of diagnosis and therapeutic strategies. The Belgian Thyroid Cancer Study Group (BTCSG) was founded in 1990. In the present study we report data collected from 1988 to 1995 in 397 patients with a differentiated (papillary and follicular) thyroid carcinoma living in the french-speaking area of Belgium. The sex ratio female/male is 3.5 and the median ages at the diagnosis, is similar (45 yrs, 12-82) in both sexes. Seven cases of thyroid cancer were registered in young patients less than 18 yrs old. Thyroid carcinoma were associated with multinodular goiter in more than 50% cases. Cancer was bilateral in 17%. Papillary histological type accounts for 84% in our series while its diagnosis was established in 45% at early clinical stages (TO-T1). These observations could probably be related with 1) broader indications and more aggressive options for the surgical removal of diffuse multinodular goiter, 2) more sophisticated pathologic examinations that might have led to the detection of a greater incidence of occult carcinomas, incidentally discovered. Lymph nodes metastases were present at the time of diagnosis in 20%, especially in young patients. The risk for local and/or lateral recurrence or distant metastases is significantly related to the size of the tumor, histologically verified lymph node metastases and the values of the EORTC prognostic index (> or = 50) that additionally takes into account the differentiation of the tumor. Considering our short median follow-up time of 25 months, it is currently too early to define if the controversial attitude about the extent of surgery (total thyroidectomy plus I131 or individualized surgery) can also negatively influence the risk for recurrence. In our series, eight patients died of thyroid cancer.

[The effect of cross-linked protein microcapsules on cell cultures]. / Effet de microcapsules de protéines réticulées sur des cultures de cellules.

Microcapsules (diameter range: 5 to 100 microns) prepared through interfacial cross-linking of proteins with terephthaloylchloride exhibited a cytotoxic effect on L 1210 cell cultures. IC50 was: 0.86 mg/ml +/- 0.24 for microcapsules prepared from human serum albumin (AT microcapsules) and 0.63 mg/ml +/- 0.05 for those obtained from egg white lysozyme (LT microcapsules). With K 562 cells IC50 were 0.42 +/- 0.11 mg/ml (AT microcapsules), 0.06 mg/ml (LT microcapsules). An increase in the cytotoxicity was observed when reducing the size of the microcapsules and when increasing the reaction pH or the terephthaloylchloride concentration, or the relative concentration of microcapsules vs cells. On the contrary, the cytotoxic effect decreased, when prolonging the cross-linking time. The activity was not affected when the microcapsules were washed with toluene or with an alkaline solute. The cytotoxic effect, which appears for relatively high doses, apparently involves a contact between the microcapsules and the cells and seems to be related with the degree of cross-linking of the constitutive protein.

[The value of screening using cervicography in dysplasia and neoplasms of the uterine cervix]. / Intérêt du dépistage par cervicographie dans la pathologie dysplasique et néoplasique du col utérin.

In cervical dysplasia and neoplasia, recent studies have reported the unreliability of the pap smear. We have used a new screening technique, cervicography and have compared it to cervix cytology. We have performed biopsies under colposcopy as reference and quality control. On a series of 65 selected patients with pap smear screening program which were sent to the colposcopic triage, the cervicography is more sensitive (83%) than the pap smear (sensitivity: 42%). However, those two tests are from a specificity point of view, comparable (cervicography: 95% versus pap smear: 98%). The association of these two techniques has detected all the lesions: they seem to be complementary. In the Bordet Institute we decided to systematically perform both pap smear and cervicography for cervix screening.