A universal UHPLC-CAD platform for the quantification of polysaccharide antigens.

Several glycoconjugate-based vaccines against bacterial infections have been developed and licensed for human use. Polysaccharide (PS) analysis and characterization is therefore critical to profile the composition of polysaccharide-based vaccines. For PS content quantification, the majority of Ultra High Performance Liquid Chromatography (UHPLC) methods rely on the detection of selective monosaccharides constituting the PS repeating unit, therefore requiring chemical cleavage and tailored development: only a few methods directly quantify the intact PS. The introduction of charged aerosol detector (CAD) technology has improved the response of polysaccharide analytes, offering greater sensitivity than other detector sources (e.g., ELSD). Herein, we report the development of a universal UHPLC-CAD method (UniQS) for the quantification and quality evaluation of polysaccharide antigens (e.g., Streptococcus Pneumoniae, Neisseria meningitidis and Staphylococcus aureus). This work laid the foundation for a universal UHPLC-CAD format that could play an important role in future vaccine research and development helping to reduce time, efforts, and costs.

Impact of fractionated stereotactic radiotherapy on activity of daily living and performance status in progressive/recurrent glioblastoma: a retrospective study.

BACKGROUND: The prognosis of recurrent glioblastoma (GBM) is poor, with limited options of palliative localized or systemic treatments. Survival can be improved by a second localized treatment; however, it is not currently possible to identify which patients would benefit from this approach. This study aims to evaluate which factors lead to a lower Karnofsky performance status (KPS) score after fractionated stereotactic RT (fSRT). METHODS: We retrospectively collected data from patients treated with fSRT for recurrent GBM at the Institut de Cancérologie de Lorraine between October 2010 and November 2017 and analyzed which factors were associated with a lower KPS score. RESULTS: 59 patients received a dose of 25 Gy in 5 sessions spread over 5-7 days (80% isodose). The median time from the end of primary radiotherapy to the initiation of fSRT was 10.7 months. The median follow-up after fSRT initiation was 8.8 months. The incidence of KPS and ADL impairment in all patients were 51.9% and 37.8% respectively with an adverse impact of PTV size on KPS (HR = 1.57 [95% CI 1.19-2.08], p = 0.028). Only two patients showed early grade 3 toxicity and none showed grade 4 or late toxicity. The median overall survival time, median overall survival time after fSRT, median progression-free survival and institutionalization-free survival times were 25.8, 8.8, 3.9 and 7.7 months, respectively. Initial surgery was associated with better progression-free survival (Hazard ratio (HR) = 0.48 [95% CI 0.27-0.86], p = 0.013). CONCLUSIONS: A larger PTV should predicts lower KPS in the treatment of recurrent GBM using fSRT.

Radiographic evaluation of cardiac silhouette using vertebral heart size method in squirrel monkey (Saimiri sciureus).

BACKGROUND: The Saimiri sciureus represents a valuable resource for biomedical research, due to its large number on primate centers and similarity to human anatomy and physiology. The aim of this study was to apply the method proposed by Buchanan & Bücheler in healthy squirrel monkeys (S. sciureus) and determine by radiographic examination, the average value of the vertebral heart size (VHS) for the specie. METHOD: Laterolateral left (LLL), laterolateral right (LLR), ventrodorsal (VD), and dorsoventral (DV) projections were used for radiographic examinations to evaluate the heart silhouette of S. sciureus. RESULTS: There was no statistical difference between the projections LLL and LLR, or VD and DV, and between males and females, so measurements in the four radiographic projections were not performed. CONCLUSION: The values obtained in VHS of S. sciureus showed up species specific.

Intraoperative identification of the negative motor network during awake surgery to prevent deficit following brain resection in premotor regions.

INTRODUCTION: Surgical resection in premotor areas can lead to supplementary motor area syndrome as well as a permanent deficit. However, recent findings suggest a putative role of the negative motor network in those dysfunctions. Our objective was to compare the functional results in two groups of adult patients who underwent the resection of a frontal glioma with and without resection of the negative motor networks. MATERIAL AND METHODS: Twelve patients (total of 13 surgeries) were selected for awake surgery for a frontal glioma. Negative motor responses were monitored during surgery at the cortical and subcortical levels. Sites eliciting negative motor responses were first identified then spared (n=8) or removed (n=5) upon oncological requirements. RESULTS: In the group with removal of the negative motor network (n=5), all patients presented a complete supplementary motor area syndrome with akinesia and mutism. At 3months, they all presented bimanual coordination dysfunction and fine movement disorders. In the group with preservation of the negative motor network (n=8), all patients presented transient and slight disorders of speech or upper limb, they all recovered completely at 3months. DISCUSSION: The negative motor network is a part of a modulatory motor network involved in the occurrence of the supplementary motor area syndrome and the permanent deficit after resection in premotor areas. Then, intraoperative functional cortico-subcortical mapping using direct electrostimulation under awake surgery seems mandatory to avoid deficit in bimanual coordination and fine movements during surgery in premotor areas.

Eurasiaplex: a forensic SNP assay for differentiating European and South Asian ancestries.

We have selected a set of single nucleotide polymorphisms (SNPs) with the specific aim of differentiating European and South Asian ancestries. The SNPs were combined into a 23-plex SNaPshot primer extension assay: Eurasiaplex, designed to complement an existing 34-plex forensic ancestry test with both marker sets occupying well-spaced genomic positions, enabling their combination as single profile submissions to the Bayesian Snipper forensic ancestry inference system. We analyzed the ability of Eurasiaplex plus 34plex SNPs to assign ancestry to a total 1648 profiles from 16 European, 7 Middle East, 13 Central-South Asian and 21 East Asian populations. Ancestry assignment likelihoods were estimated from Snipper using training sets of five-group data (three Eurasian groups, East Asian and African genotypes) and four-group data (Middle East genotypes removed). Five-group differentiations gave assignment success of 91% for NW European populations, 72% for Middle East populations and 39% for Central-South Asian populations, indicating Middle East individuals are not reliably differentiated from either Europeans or Central-South Asians. Four-group differentiations provided markedly improved assignment success rates of 97% for most continental Europeans tested (excluding Turkish and Adygei at the far eastern edge of Europe) and 95% for Central-South Asians, despite applying a probability threshold for the highest likelihood ratio above '100 times more likely'. As part of the assessment of the sensitivity of Eurasiaplex to analyze challenging forensic material we detail Eurasiaplex and 34-plex SNP typing to infer ancestry of a cranium recovered from the sea, achieving 82% SNP genotype completeness. Therefore, Eurasiaplex provides an informative and forensically robust approach to the differentiation of European and South Asian ancestries amongst Eurasian populations.

Magnetic resonance hysterosalpingography in the evaluation of tubal patency in infertile women: an observational study.

The purpose of this study was to evaluate the ability of magnetic resonance hysterosalpingography (MR-HSG) to demonstrate fallopian tube patency in infertile women and to improve the MR-HSG technique. Sixteen consecutive infertile women were recruited for this trial. All subjects underwent clinically indicated MR-HSG: 10-15 ml of 1:10 solution of gadolinium and normal sterile saline (0.9%) was gently hand-injected intracervically through a 7 French balloon catheter while seven consecutive flash-3D dynamic (FL 3D DY) T1-weighted MR sequences were acquired. Two readers independently assessed image quality as well as anatomic and pathologic correlations. Patient comfort was evaluated using a specific score questionnaire. MR-HSG was successfully completed in all patients. In 14/16 (87.4%) patients, MR-HSG showed bilateral tubal patency with symmetric contrast agent diffusion and a regular tubo-ovarian relationship. One patient (6.3%) with monolateral hydrosalpinx presented no contrast agent diffusion in the affected side (monolateral tubal occlusion); in another patient (6.3%) the fallopian tube was displaced upward causing loss of the tubo-ovarian anatomical relationship resulting in asymmetric and inadequate contrast agent diffusion. Eight women (50%) were found to have abnormalities on MR imaging; these abnormalities included multi follicular ovaries (5 cases 31.1%), endometrioma (1 case, 6.3%), leiomyoma (1 case/6.3%) and endometrial polyp (1 case/6.3%). The average time required for the study was 25-30 minutes. Analysis of the questionnaires administered to the patients showed that 15/16 patients (93.7%) were fully satisfied with the procedure. All examinations were judged to be of high diagnostic quality and the two readers made similar diagnoses. In conclusion, MR-HSG is a feasible, useful and well tolerated tool for the assessment of the uterus, fallopian tubes, ovaries and extra-uterine structures. MR-HSG is a new promising imaging approach to female infertility.

Mammographic features in infertile women as a potential risk for breast cancer: a preliminary study.

The purpose of the present study was to evaluate breast mammographic features, particularly mammographic density in a selected population of infertile women and to assess if these women should be considered at higher risk for breast cancer. The prevalence of female infertility in Western countries is approximately 10-15% and since causes affecting the female are involved in 35-40%, concerns have developed about the future health of these women, specifically whether infertility could represent a risk factor for future cancer development. Moreover, infertility is now often treated with medication and procedures that could modify the hormonal environment and be cofactors in the cellular changes towards cancer development. Mammographic breast density is a useful marker for breast cancer risk and breast density is considered one of the strongest risk factors for breast cancer. Breast density is associated with known breast cancer risk factors such as reproductive and menstrual factors including serum estrogen and progesterone concentrations. In Italy the National Federation for Breast Cancer (FONCAM) guidelines suggest the usefulness of mammography from 35 years of age for women who undergo infertility hormone therapy (FONCAM Guidelines, 2005). According to this recommendation 294 women aged > or = 35, with primary infertility, sent to our breast service before joining an IVF program were recruited and then underwent clinical examination and X-ray mammography. Women were divided into two groups: dense breast (DB) and non-dense breast (NDB). Univariate analysis was employed to evaluate if there was an association between mammographic density and other risk factors. Evaluation of mammographic features showed the presence of BI-RADs C and D in the sample of 200 (68%) patients with DB and in 94 (32%) patients with NDB BI-RADS A and B. Univariate analysis showed that there were no statistically significant differences between the groups BD and NDB as regards age at mammography, age at menarche, BMI and family history for breast cancer, while ovulatory etiology of infertility was found to be associated with high mammographic density (p < 0.05). In conclusion, bearing in mind that 68% of our study sample had high breast density, we can assume that patients with primary infertility might represent a group at high risk for breast cancer, particularly if infertility is due to an ovulatory factor. We suggest breast screening from the age of 35 in infertile patients who undergo treatment with fertility drugs in accordance with FONCAM recommendations. This might allow the identification of higher risk patients who need more closely monitored breast examinations.

[Cervical cerclage and evidence-based medicine: if, how and when]. / Il cerchiaggio cervicale e la medicina basata sull'evidenza: se, come e quando.

Cervical cerclage has always been the main treatment option in cases of so-called cervical insufficiency, a condition that is notoriously associated with a high risk of second trimester abortion and/or preterm delivery. We can distinguish between a prophylactic cerclage, to be performed electively, usually at 13-16 weeks gestation, only when the woman has a history extremely suggestive for cervical incompetence (3 or more mid-trimester abortions or preterm deliveries) and a therapeutic cerclage. This last cerclage is recommended either for women who have ultrasonographic changes consistent with a short cervix or the presence of funneling after the 16-20 weeks gestation (urgent cerclage) and for women who present the asymptomatic dilation of the uterine cervix of at least 2 cm and/or a prolapse of the amniochorial membranes (emergent cerclage). So far there is still a lack of controlled and randomized trials that can unquestionably demonstrate the advantages of the cervical cerclage in comparison with a ''wait and see'' aptitude. The cerclage can be performed either transvaginally, usually according to the McDonald technique, or transabdominally. This last approach is recommended when a transvaginal cerclage has to be avoided because of technical difficulties depending on the conditions of the cervix or when the pregnant woman has a history of one or more failed transvaginal cerclages. Interesting perspectives are currently offered by the laparoscopic cerclage, a method that has been effective and unexpectedly safe till now.

Valoración médico forense del esguince cervical / Medico legal evaluation of whyplash injury

El esguince cervical es la patología traumática más frecuente en el ámbito de la valoración médico forense. Se realiza una aproximación a la misma a través de las peritaciones realizadas por 22 médicos forenses de 15 partidos judiciales españoles, sobre un total de 572 lesionados. Se concluye la mayor incidencia en el sexo femenino (57.52%), en el rango de edades entre 21 y 40 años (58%), siendo conductor de un turismo (54.7%) que recibe un golpe posterior (59%). Con un tiempo de sanidad entre los 31 y 90 días (52%), resultando en un mayor porcentaje con secuelas (64,3%), comúnmente SPC o cervicalgia, generalmente (56%) de grado leve

Whiplash is the leading traumatic cause concerning the evaluation in forensic activity. It has been carried out an approximation through the reports done by 22 forensic examiners working in 15 Spanish jurisdictions, over a total amount of 572 injured people. In fact there is a bigger incidence of damage in female gender (57.52%), in the age ranging from 21 to 40 years (58%), driving a car (54.7%), following a rear-end collision (59%). The healing time varies from 31 to 90 days (52%), and mostly with sequels (64.3%), commonly Cervical Post-Traumatic Syndrome or cervical pain, usually (56%) in a low degree

[Placental chorioangioma]. / Il corionangioma placentare.

Chorioangioma is a primary benign tumour with predominantly vascular involvement. It is found in 1% of all placentas undergoing careful and systematic histopathological examination. A rarer form is the clinically relevant chorioangioma which is likely to be associated with complications like polyhydramnios, pre-term birth, IUGR, anemia, fetal congested heart decompensation, non-immune fetal hydrops, perinatal mortality. Chorioangioma generally presents an increased consistency compared to adjacent tissues; it tends to be single, rounded and reddish-brown in colour. The preferred localisation is on the fetal side of the placental disk and it may protrude into the amniochorial cavity; the volume is variable (but rarely exceeds a maximum diameter of 4-5 cm). Of the known 3 histotypes of chorioangioma (angiomatous, cellular, degenerative), the first is the most insidious: the intratumoral vascular bed represents an arteriovenous shunt in the systematic circulation of the fetus #150; sometimes causing dramatic hemodynamic consequences #150; as well as a functional empty space (blood short-circuited through the neoplasm circulation would be deprived of the possibility of adequate gaseous and metabolic exchanges at the level of the terminal villi). Having been identified as early as 1798, chorioangioma has recently been the subject of renewed interest for the following reasons: 1) the possibility of prenatal diagnosis, prohibited to earlier generations of authors, following the development of instrumental techniques like ultrasonography and flowmetry; 2) the acquisition of new knowledge regarding the physiopathology of the tumour and the etiopathogenesis of its main complications; 3) improved prognostic capacity accompanied by better prospects for correct management; 4) wider awareness of the range of therapeutic options available.

Optimization of parameters for the supercritical fluid extraction in the determination of N-nitrosamines in rubbers.

The study of the possibilities of supercritical fluid extraction (SFE) with N-nitrosamines in rubbers has been carried out. Home-made materials fortified with several N-nitrosamines were prepared in order to optimize the SFE parameters. A Plackett-Burman design was employed to evaluate the influence of those parameters to be controlled in SFE, such as pressure, temperature, static and dynamic time, restrictor temperature and volume of modifier while CO2 was used as the extraction fluid. An extra central composite design for the main factors (according to the previously obtained results) was also developed in order to refine the best supercritical conditions for the extraction of N-nitrosamines from rubbers. Gas chromatography with a nitrogen and phosphorus sensitive detector was used to achieve sensitivity and limits of detection for the concentrations expected in plastic materials. The proposed analytical method has shown to be useful in the determination of N-nitrosamines even for complex matrices.

Improvement of the accuracy by the measurement of the electrical cell membrane parameters.

The electrical parameters of the cell membrane are mostly estimated employing ac methods. The measurement is based on the analysis of the current(s) flowing through an access resistance and the membrane. A current/potential transducer is used at the input of the device. The parameters of this transducer, especially its feedback capacity, degrades the accuracy of the measurement and hence diminishes the suppression of mutual influences of the individual parameters. The paper suggests a possible software correction and is supplemented by remarks for practical application.

[PGF2 and the induction of the trial of labor. The prognostic value of uterine contractions]. / PGE2 e induzione del travaglio di parto. Valore prognostico delle contrazioni uterine.

BACKGROUND: Uterine contractions during the first hour following intracervical application of dinoprostone would show the myometrial sensitivity to prostaglandins E2 (PGE2) and could be a good practical marker of the real prospects of success of an attempt to induce labor according to the above mentioned modalities. The verification of such an hypothesis is the principal aim of this work. METHODS: The study was carried out on a group of 90 cardiotocograms recorded soon after a sample of pregnant women at term underwent labor induction by intracervical application of a gel containing 0.5 mg of PGE2 (dinoprostone). Special attention was paid to some characteristics of the cardiotocograms obtained during the first hour following administration of the gel: regularity of uterine contractions, total number of contractions, number of contractions having an intensity equal or superior to 50 mmHg, mean number of contractions during a period of 10 minutes, mean number of "effective" contractions during a period of 10 minutes, presence or absence of "excessive uterine activity" (tachysystole and/or hypersystole). RESULTS: Some cardiotocographic patterns were associated with a higher percentage of successful inductions, but the observed differences, not statistically significant, do not seem to be clinically interesting. CONCLUSIONS: However, it's not possible to exclude that a longer period of uterine contractions monitoring, perhaps lasting 2 hours instead of only 60 minutes, could lead to more useful information for the management choices.

[Collection of autologous blood during pregnancy and fetal safety. Computer analysis of fetal heart rate]. / Predonazione di sangue autologo in corso di gravidanza e sicurezza fetale. Analisi computerizzata dell'attività cardiaca fetale.

BACKGROUND: This study aims at objectively evaluating the impact on the fetus of a sudden maternal blood withdrawal, through a computer analysis of the various aspects of the cardiotocographic pattern before and after one autologous blood unit predonation from near term pregnant women. METHODS: For the collection of cardiotocographic data, a Sonicaid Team fetal monitor was used; it is a monitoring system which uses Doppler ultrasound impulses and autocorrelates the received Doppler signals with varied signals of frequency, connected to Sonicaid System 8000, a program for "objective" cardiotocographic analysis according to the so-called Dawes-Redman criteria. RESULTS: From the statistic comparison of Non Stress Tests preceeding and following predonation, some statistically significant differences have sometimes emerged on the level of specific parameters. CONCLUSIONS: In particular, the general variation of fetal heart frequency and fetal reactivity resulted higher after predonation, even though they were within absolute normality, justifying the suspicion that the stress of autologous predonation practice might produce some indefinite disturbance to fetal activity/rest rhythm.

[Predictive factors of fetal macrosomia]. / Fattori predittivi di megalosomia fetale.

BACKGROUND: Fetal macrosomia is a condition which may increase the risk of mechanical and/or dynamic problems for the parturient. In the past, we have demonstrated that in more than half of the cases it is not possible to exclude a contribution of maternal pathology to the determination of fetal-macrosomia. The aim of this work is to verify whether our more recent experience regarding predictive factors of fetal macrosomia shows some noteworthy novelty. METHODS: The study was retrospectively carried out on pregnant women who, during the period January 1994-February 1996, delivered babies weighing at least 4 kg at the Midwifery School of Camerino. With regard to the frequency of the main risk factors of fetal macrosomia described in scientific literature, the sample was compared with a control group randomly selected. RESULTS: Advanced gestational age at the time of delivery, parental tallness, maternal overweight/obesity, gestational glucidic dysmetabolism, a distance between pubis and uterine fundus of at least 34 cm, male sex of the unborn child have proved to be predictive factors of fetal macrosomia. CONCLUSIONS: The differences in comparison with the past, on the other hand altogether negligible, are the consequence of changes in the management of some obstetric situations.

[The role of chorionic gonadotropin in transient hyperthyroidism in hyperemesis gravidarum]. / Il ruolo della gonadotropina corionica nell'ipertiroidismo biochimico transitorio dell'iperemesi gravidica.

A possible association of hyperemesis gravidarum with biochemical transient hyperthyroidism (a significant self-limited increase in serum levels of some thyroid hormones) has long been reported. It seems there is not any causal connection between the two above mentioned conditions, but they may be both independently related to the same cause: the trophoblastic production of a large amount of human chorionic gonadotropin or, otherwise, the presence in the maternal circulation of structural variants of hCG with higher biological activity. The validity of such hypothesis, denied by some authors, encounters the verification in the peculiar clinical case described in this report. It also shows clearly the danger of considering a case of hyperemesis gravidarum as of psychological nature, without having preliminarily excluded an organic cause. For the pregnant woman, to hear of her unconscious refusal of pregnancy or of the timeliness of her removal from the environment in which some conflictual situations have likely raised or, furthermore, to hear of other common places often too easily evoked could be cause of anxiety and unjustified feelings of guilt.

[PGE2 and induction of labor. Authors' experience]. / PGE2 e induzione del travaglio di parto. Nostra esperienza.

BACKGROUND: The aim of this work is to contribute to the evaluation of the dinoprostone clinical value (PGE2) in labor induction. Special attention is paid to the evaluation of the effectiveness and safety of the method employed, as well as to the singling out of possible predictive factors of success. METHODS: The study was carried out on a sample of 85 women, who underwent labor induction by intracervical application of a gel containing 0.5 mg of dinoprostone. The following parameters were evaluated: 1) percentage of spontaneous deliveries within 24 hours from the administration of dinoprostone; 2) kind of delivery (comparison with women who delivered without labor induction); 3) characteristics of the cardiotocogram performed soon after the application of the gel; 4) prognostic value of some factors (maternal age, parity, gestational age, Bishop score). RESULTS: The difference between the two categories compared according to susceptibility of a positive answer to the treatment with PGE2 is mainly the consequence of different Bishop scores. The Bishop score was the only examined having an autonomous prognostic value and a real indicator of success prospects of an attempt to induce, labor by administration of PG. CONCLUSIONS: In personal experience, the una tantum intracervical application of dinoprostone is an effective method and, at the same time, it is free from any major risk.

[Breast feeding. The importance of correct management]. / Allattamento al seno. L'importanza di un corretto management.

The advantages that breast-feeding still offers over bottle-feeding are widely recognized. This study has the following aims: 1) to verify how many of the women that delivered during the last biennium at the Midwifery School of Camerino breastfed their infants for a fair amount of time; 2) to evaluate the relationship between the results obtained and the protocols followed in the management of the various issues connected with breast-feeding. The research was carried out into a sample of 140 women, randomly selected. The anamnestic and clinical data obtained from the medical cards were integrated with information gathered by telephonic interviews. Less than 40% of the women of the sample gave their infants only maternal milk for a very good period. The inobservance of many WHO guidelines for the promotion of breast-feeding seems to be the main cause of the disappointing results obtained. Some groups of women proved more exposed to the risk of inadequate suckling: teenagers, the older gravidae, women that delivered prematurely, those that had children with a low birth weight, the women that had previous breast-feeding failures, smokers, women that attended a course of psychoprophylaxis, women with the lowest socioeconomic standards, mothers that were precociously separated from their babies, women that had some gestational pathology or some problems during the first days after delivery, women that had as main reference doctors without a proper breast-feeding culture.

A method of resolution improvement by the measurement of cell membrane capacitance.

When measuring cell membrane electrical capacitance in whole cell configuration using alternating currents, the resolution decreases with increasing membrane conductance and pipette resistance. Improved resolution was attained by the dual-frequency method which was modified as to control the voltage amplitude of one of the measuring frequencies. A model circuit was developed for the verification of the method. This circuit allows measurement of calibrated capacitance changes even in the range of 5 to 20 fF. Moreover, the method was applied to capacitance measurements on pancreatic exocrine acinar cells. The results of measurements on the model as well as on pancreatic acinar cells are presented. The principle can also be applied to other hardware and software methods for measuring electrical cell membrane parameters.