Oral Vaccination with Hepatitis E Virus Capsid Protein and Immunobiotic Bacterium-Like Particles Induce Intestinal and Systemic Immunity in Mice.

The hepatitis E virus (HEV) genotype 3 (GT3) is an emergent pathogen in industrialized countries. It is transmitted zoonotically and may lead to chronic hepatitis in immunocompromised individuals. We evaluated if the major antigen of HEV, the capsid protein, can be used in combination with immunobiotic bacterium-like particles (IBLP) for oral vaccination in a mouse model. We have cloned and expressed the RGS-His5-tagged HEV GT3 capsid protein (ORF2) in E. coli and purified it by NiNTA. IBLP were obtained from two immunobiotic Lactobacillus rhamnosus strains acid- and heat-treated. ORF2 and the IBLP were orally administered to Balb/c mice. After three oral immunizations (14-day intervals), blood, intestinal fluid, Peyer´s patches, and spleen samples were drawn. IgA- and IgG-specific antibodies were determined by ELISA. Mononuclear cell populations from Peyer's patches and spleen were analyzed by flow cytometry, and the cytokine profiles were determined by ELISA to study cellular immunity. Orally administered recombinant ORF2 and IBLP from two L. rhamnosus strains (CRL1505 and IBL027) induced both antigen-specific humoral and cellular immune responses in mice. IBLP027 was more effective in inducing specific secretory IgA in the gut. IFN-γ, TNF-α, and IL-4 were produced by Peyer's plaques lymphocytes stimulated with ORF2 ex vivo suggesting a mixed Th1/Th2-type adaptive immune response in immunized mice. Oral vaccines are not invasive, do not need to be administered by specialized personal, and elicit both systemic and local immune responses at the port of entry. Here, we present an experimental oral vaccine for HEV GT3, which could be further developed for human and/or veterinary use.

Are there intracellular Ca2+ oscillations correlated with flagellar beating in human sperm? A three vs. two-dimensional analysis.

STUDY QUESTION: Are there intracellular Ca2+ ([Ca2+]i) oscillations correlated with flagellar beating in human sperm? SUMMARY ANSWER: The results reveal statistically significant [Ca2+]i oscillations that are correlated with the human sperm flagellar beating frequency, when measured in three-dimensions (3D). WHAT IS KNOWN ALREADY: Fast [Ca2+]i oscillations that are correlated to the beating flagellar frequency of cells swimming in a restricted volume have been detected in hamster sperm. To date, such findings have not been confirmed in any other mammalian sperm species. An important question that has remained regarding these observations is whether the fast [Ca2+]i oscillations are real or might they be due to remaining defocusing effects of the Z component arising from the 3D beating of the flagella. STUDY DESIGN, SIZE, DURATION: Healthy donors whose semen samples fulfill the WHO criteria between the age of 18-28 were selected. Cells from at least six different donors were utilized for analysis. Approximately the same number of experimental and control cells were analyzed. PARTICIPANTS/MATERIALS, SETTING, METHODS: Motile cells were obtained by the swim-up technique and were loaded with Fluo-4 (Ca2+ sensitive dye) or with Calcein (Ca2+ insensitive dye). Ni2+ was used as a non-specific plasma membrane Ca2+ channel blocker. Fluorescence data and flagella position were acquired in 3D. Each cell was recorded for up to 5.6 s within a depth of 16 microns with a high speed camera (coupled to an image intensifier) acquiring at a rate of 3000 frames per second, while an oscillating objective vibrated at 90 Hz via a piezoelectric device. From these samples, eight experimental and nine control sperm cells were analyzed in both 2D and 3D. MAIN RESULTS AND THE ROLE OF CHANCE: We have implemented a new system that allows [Ca2+]i measurements of the human sperm flagellum beating in 3D. These measurements reveal statistically significant [Ca2+]i oscillations that correlate with the flagellar beating frequency. These oscillations may arise from intracellular sources and/or Ca2+ transporters, as they were insensitive to external Ni2+, a non-specific plasma membrane Ca2+ channel blocker. LARGE SCALE DATA: N/A. LIMITATIONS REASONS FOR CAUTION: Analysis in 3D needs a very fast image acquisition rate to correctly sample a volume containing swimming sperm. This condition requires a very short exposure time per image making it necessary to use an image intensifier which also increases noise. The lengthy analysis time required to obtain reliable results limited the number of cells that could be analyzed. WIDER IMPLICATIONS OF THE FINDINGS: The possibility of recording flagellar [Ca2+]i oscillations described here may open a new avenue to better understand ciliary and flagellar beating that are fundamental for mucociliary clearance, oocyte transport, fertilization, cerebrospinal fluid pressure regulation and developmental left-right symmetry breaking in the embryonic node. STUDY FUNDING AND COMPETING INTEREST(S): This work was supported by Consejo Nacional de Ciencia y Tecnología (CONACyT) (grants 253952 to G.C.; 156667 to F.M.M. and Fronteras 71 39908-Q to A.D. and Post-doctoral scholarships 366844 to P.H.-H. and 291028 to F.M.) and the Dirección General de Asuntos del Personal Académico of the Universidad Nacional Autónoma de México (DGAPA-UNAM) (grants CJIC/CTIC/4898/2016 to F.M. and IN205516 to A.D.). There are no conflicts of interest to declare.

[Not Available]. / Ictère nucléaire dans un contexte de naissance à domicile.

This is the clinical history of a term baby born at home who presents a severe hyperbilirubinémia. The medical monitoring was assessed by a private midwife according to parental choice. On the third day of life, the newborn presented an icterus and was exposed to natural daylight in the familial greenhouse under the midwife recommandations. On that day, no laboratory test precised the bilirubin level. On the fifth day, a blood sampling revealed a very high blood bilirubinémia (31 mg/dl or 527 mmol/L), the baby is refered to our NICU and underwent an exchange transfusion. The radiological assessment report structural abnomalies in basal ganglia seen on both MRI and transfontannellar echography. These lesions are known to be responsible of cerebral palsy and hearing loos. The neurophysiologic investigations showed background abnormaly and depression. The extensive blood sampling excluded haemolysis. The clinical examination brought out neurologic impairement and weight loos in this exclusively breastfed baby. This clinical case point out the increasing risk of home Kernicterius as hospital stays diminish and homebirth enthousiasm rise up. The present clinical situation vouches for an adaptation of care giving to both mother and child at home in order to avoid this severe illness.

[Systematic follow-up of infants born preterm]. / Le suivi systématique des grands prématurés.

Advances in pediatric medicine have enabled a decrease in perinatal mortality, especially among infants born preterm (< 32 weeks gestational age) or low birth weight (< 1.500 g). However, this population is exposed to a greater risk of neurological sequelae. This is why the creation of specific follow-up program are mandatory to screen at-risk children to offer them a support able to minimize the impact of prematurity on their future neurological development.

[Patient with a Fontan circulation undergoing caesarean section: Anesthesiological management]. / Sectio caesarea bei einer Patientin mit Fontan-Zirkulation : Anästhesiologisches Management.

Adults suffering from congenital heart diseases (CHD) represent a challenge to anesthesiologists because of the diverse pathologies, complex pathophysiology and special treatment strategies. Due to improved therapeutic options for CHD, patient quality of life and life expectancy is increasing, leaving them as a growing population including pregnant patients with CHD. This article presents the main principles of the pathophysiology and anesthesiological management of pregnant patients living with a Fontan circulation based on a case report, which was complicated by an aortic coarctation and atonic uterine hemorrhage.

[Long-term visual outcome comparison of bilateral implantation of apodised diffractive versus progressive multizonal refractive multifocal intraocular lenses after cataract extraction]. / Langzeitergebnisse nach Implantation apodisiert diffraktiver versus progressiv refraktiver Multi-Zonen-Intraokularlinsen nach Kataraktoperation.

AIM: The aim of this study was to evaluate the functional effect of bilateral implantation of apodised diffractive versus progressive multizonal refractive multifocal intraocular lenses compared to standard monofocal intraocular lenses. PATIENTS AND METHOD: 229 patients underwent cataract extraction; 66 Array SA40N (AMO, Irvine, CA, USA), 76 SA60D3 ReSTOR (Alcon, Fort Worth, TX, USA), and 87 MA60AC (Alcon, Fort Worth, TX, USA) were implanted. In this retrospective trial the main outcome measures were near, intermediate, and distance visual acuity and assessment of subjective function by questionnaire. RESULTS: Mean follow-up was 4.7 ± 1.4 years with monofocal, 6.6 ± 1.7 with Array and 4.3 ± 1.1 with ReSTOR implants. Uncorrected binocular distance visual acuity was equivalent in the three groups. Uncorrected binocular near and intermediate visual acuity and spectacle independence were significantly higher in the two multifocal groups (p < 0.001). Glare and halos were more bothersome with multifocal than monofocals implants (p < 0.05) and adverse visual symptoms at night with Array implants but equivalent between ReSTOR patients and monofocal patients. Between the two multifocal groups spectacle independence was higher and adverse visual symptoms lower in ReSTOR patients than in Array patients (p < 0.05). ReSTOR patients reported a higher overall visual satisfaction than the other groups (p < 0.001) and rated their vision at 8.8 ± 1.8. Monofocal patients reported a slightly higher satisfaction at 7.6 ± 1.7 compared to Array patients at 6.9 ± 2.6 (p = 0.05). CONCLUSION: In this long-term study the highest overall visual satisfaction could be achieved by bilateral implantation of apodised diffractive intraocular lenses.

[Neurodevelopment of extremely low birthweight infants born in Erasmus Hospital between 1992 and 2001]. / Suivi neurodéveloppemental des prématurissimes nés a l'Hôpital Erasme entre 1992 et 2001.

Survival of extremely low birth weight infants has dramatically improved in the last decade of the twentieth century. The objective of our study was to evaluate the neurological evolution of the surviving infants because frequent motor, sensitive and psychological disturbances are related. Prospective, longitudinal study in a population of newborns, nursed in our neonatal intensive care unit and born between 1992 and 2001 with less than 1.000 g and/or less than 28 weeks of gestational age (GA). Neurological assessment of outcome was made using the neurodevelopmental score (O.M.S. 1988) at 6, 9, 12, 18, 24 and 36 months. Neurological follow-up every year and neuropsychological testing at 3, 5 and 8 years. Only children with at least 2 years of follow-up were included. The children were grouped in 3 categories: M (major neurological handicap), m (minor neurological handicap), N (normal neurological outcome). To evaluate the evolution with time, we compared the results from the first period (1992 to 1996) to the second part of this decade (1997 to 2001). Mortality fell from 38% (27/70) in the first period (1992-1996) to 18% (8/44) in the second one (1997-2001) (p = 0.02) including neonates of less than 25 weeks GA. Neurodevelopmental status improved and severe brain lesions decreased (25% with intraventricular haemorrhage III & IV and cystic periventricular leukomalacia versus 6% in the second period) (p = 0.017). Major handicap fell from 26% (9/34) to 16% (5/31) and normal neurological evolution raised from 15% (5/34) to 48% (15/31) (p = 0.013). With the survival of newborns less than 28 weeks, the severe ocular complications increased: 6% (5/79). In conclusion, mortality and quality of life have significantly improved in the past 10 years in our service. Severe brain lesions have decreased under a better multifactorial management. Nevertheless when the gestational age of the surviving babies diminishes, ocular sequelae increase. We still think that prematurity remains a burden for the child, his family and the society.

[Papillary hemodynamics in patients with normal pressure glaucoma and hemorrhage of the optic papilla circumference]. / Papilläre Hämodynamik bei Patienten mit Normaldruckglaukom und Papillenrandblutungen.

BACKGROUND: Optic disc hemorrhages in patients with normal-pressure glaucoma (NPG) are usually regarded as a sign of vascular dysfunction and as an indicator for glaucoma damage progression. METHODS: Optic nerve head blood flow was measured in 21 patients suffering from NPG with acute optic disc hemorrhages by scanning laser Doppler flowmetry at various locations of the optic disc. Intraocular pressure and mean deviation of the visual field were also monitored. Two groups served as control: 21 patients with NPG matched for age, sex, and stage of the disease and in addition the contralateral eye without any hemorrhages. RESULTS: Optic nerve head blood flow as a mean of several locations was significantly lower in eyes with optic disc hemorrhages than in controls and differed significantly from the contralateral eye. CONCLUSION: Optic nerve blood flow was lower in NPG eyes with optic disc hemorrhages than in the contralateral eye and in controls.

Mural aortic thrombi: An important cause of peripheral embolization.

PURPOSE: Arterial thromboembolism in patients with an unknown source of embolization is still associated with significant morbidity and mortality. The advent of transesophageal echocardiography (TEE) and magnetic resonance imaging (MRI) and the more frequent use of computed tomography (CT) have led to the identification of mural aortic thrombi (MAT) as a source of distal embolization in a much higher proportion of patients than previously appreciated. The incidence, diagnosis, and treatment of patients with MAT is reported. METHODS: In a prospective study, from January 1996 to December 1998, 89 patients with acute embolic events underwent an extensive diagnostic workup, consisting of TEE, CT, or MRI, to detect the source of embolization. Patients in whom the heart (n = 51), occlusive aortoiliac disease (n = 16), or aortic aneurysms (n = 12) was identified as the source of embolization were excluded. RESULTS: Five female and three male patients, with a median age of 63 years (range, 35 to 76 years), with bilateral or repetitive embolic events resulting from MAT were identified, representing 9% of all patients with arterial thrombembolism. All patients had several risk factors for atherosclerosis, but only one young patient had a single risk factor that promoted thrombosis. Successful percutaneous catheter aspiration embolectomy was performed in six patients. The remaining two patients underwent surgical thromboembolectomy. A below-knee amputation had to be performed in two patients, thus representing a morbidity of the primary treatment of 25%. MAT of equal value were detected in the ascending (n = 1) and thoracic aorta (n = 3) by means of TEE, CT, or MRI. MAT in the abdominal aorta (n = 4) were identified by means of CT and MRI. Surgical removal of MAT was performed in seven patients by means of graft replacement of the ascending aorta (n = 1), open thrombectomy of the descending aorta (n = 2), and thrombendarterectomy of the abdominal aorta (n = 4), without intraoperative or postoperative complications. No recurrence of MAT occurred during a median follow-up period of 13 months (range, 4 to 24 months). CONCLUSION: MAT represent an important source of arterial thrombembolism. A diagnostic workup of the aorta, preferably by means of CT or MRI, should be performed in all patients in whom other sources of embolization have been ruled out. The ideal therapeutic approach to these patients still awaits prospective evaluation. However, based on our experience, MAT can be successfully treated with a definitive surgical procedure in selected patients, with low mortality and morbidity.

Evaluation of coronary bypass flow with color-Doppler and magnetic resonance imaging techniques: comparison with intraoperative flow measurements.

OBJECTIVES: After coronary artery bypass surgery, patency and flow assessment is based on invasive methods such as angiography and intravascular ultrasound or flow wire techniques. The aim of the study was to compare intraoperative transit time flow measurements of coronary bypass grafts with early postoperative color-Doppler and MR-imaging assessment. METHODS: In 22 patients (62+/-8.5 years) undergoing elective coronary bypass surgery the flow was measured in all internal mammary artery grafts (IMA) and saphenous vein grafts using the transit time flow technique. Postoperatively (days 5-7) all patients had a color-Doppler IMA graft assessment followed by a MR-angiography and flow measurement (navigator echo phase contrast technique with and without contrast bolus application) to determine patency and graft flow. RESULTS: Data are expressed as the mean +/- SD). (1) In all patients the left IMA graft to the left anterior descending coronary artery (LAD) could be identified and flow could be assessed with both color-Doppler and MRI. Venous grafts could only be visualized by MRI. The use of an intravenous contrast bolus enhanced the visualization of coronary artery bypass grafts. (2) The mean IMA to LAD flow was 33+/-17 ml/min intraoperatively by transit time and postoperatively 36+/-25 ml/min by MR respectively 66+/-54 ml/min by color-Doppler technique. (3) The systolic/diastolic flow ratio was 0.44+/-0.12 intraoperatively and 0.43+/-0.17 postoperatively by MR respectively 0.67+/-1.0 by color-Doppler. (4) A statistically significant correlation could be demonstrated between intraoperative transit time and postoperative MR flow measurements (r = 0.57; P < 0.04), whereas the correlations to color-Doppler flow were poor. Postoperatively MR and color-Doppler showed a good correlation of systolic/diastolic flow ratio (r = 0.88; P < 0.008). CONCLUSIONS: The color-Doppler method during echocardiography and MR-imaging are useful non-invasive techniques to visualize postoperative IMA grafts for patency assessment. The quantification of IMA flow is still difficult with either technique, but MR flow measurements showed the best correlation to the intraoperatively measured transit time flow. The MR technique is the most promising non-invasive method for postoperative evaluation of coronary bypass grafts, since it allows visualization and reliable flow quantification.

[Periventricular leukomalacia in premature infants: prognostic role of ultrasonography and MRI]. / Leucomalacie périventriculaire du prématuré: rôle pronostique de l'échographie et de l'IRM.

PURPOSE: To evaluate the role of cranial US and MRI to establish the neurological prognosis of premature infants with periventricular leukomalacia (PVL). PATIENTS AND METHODS: Follow-up results of cranial US and early MRI evaluation (before 25 weeks*) of 28 premature infants were retrospectively reviewed and compared to the neurological outcome at 18 months* (*corrected age). RESULTS: Follow-up by cranial US was more sensitive (8/28) than early MRI to detect cystic PVL lesions because of the transient nature of these cysts. This has prognostic implications since all patients (8/8) with cystic PVL lesions had neurological sequelae. MRI was useful, as a complement to cranial US, for the evaluation of non-cystic PVL lesions. Indeed, patients with evidence of hemorrhage or paucity of white matter at MRI had a higher risk of neurological sequelae (9/11) than infants with echogenic periventricular white matter at US without evidence of white matter abnormality at MRI (p < 0.013). CONCLUSION: MRI was useful, as a complement to cranial US, to evaluate the prognosis of infants with non-cystic PVL lesions.

Assessment of cardiac rejection by MR-imaging and MR-spectroscopy.

BACKGROUND: Detection of cardiac rejection is a major problem in cardiac transplantation. The gold standard is, and remains, endomyocardial biopsy. PURPOSE: Evaluation of MR-imaging and MR-spectroscopy for detection of cardiac rejection. METHODS: Orthotopic cardiac transplantation (HTX) was performed in 13 pigs (body weight 30 kg). All animals obtained immunosuppressive (triple) therapy for 1 week after the operation. Thereafter immunosuppression was stopped to induce cardiac rejection. MRI and MRS (1.5 Tesla General Electrics Signa) were performed pre- and post-operatively on days 10, 17, 24 and 31. The degree of rejection was determined post-operatively using endomyocardial biopsy (Texas grading score). RESULTS: (1) MR-imaging: LV function remained unchanged after HTX. LV mass increased (+42%; P < 0.05) with cardiac rejection. (2) MR-spectroscopy: a marked reduction in the ratio of phosphocreatine and adenosine triphosphate, respectively, to inorganic phosphate was observed in the rejecting hearts. (3) Histologic grading confirmed cardiac rejection after stopping immunosuppression. The Texas score was 5.7+/-0.8 at autopsy. CONCLUSIONS: MR-imaging and MR-spectroscopy allow the detection of changes associated with cardiac rejection. Both techniques are correlated with histologic rejection. However, endomyocardial biopsy remains the gold standard for reliable detection of cardiac rejection.

[The significance of magnetic resonance venography in the pre-interventional clarification of a malignant superior venous obstruction]. / Bedeutung der Magnetresonanz-Venographie in der präinterventionellen Abklärung der malignen oberen zentralvenösen Obstruktion.

PURPOSE: Comparison between peripheral digital subtraction venography (DSV) and magnetic resonance venography (MRV) in planning for stent placement in malignant superior central venous obstruction. METHOD: 19 patients with malignant central vein obstruction were examined by DSV (n = 19) and MRV (n = 19). For each patient 12 segment-vessel evaluation was performed to review for obstruction and then compared with the gold standard--selective catheter phlebography--performed at the time of stent insertion. Additionally, tumor extension and collateral venous outflow was noted. RESULTS: Sensitivity, specificity, and accuracy amounted to 92%, 96%, and 95% for DSV and 98%, 100%, and 100%, respectively for MRV. The McNemar test revealed a significance between MR venography and DSV. CONCLUSIONS: With regard to planning of stent placement, a more exact evaluation was possible with MR venography than with DSV. Thus, MR venography can replace DSV as the method of choice for clarifying malignant superior central vein obstructions prior to stent placement.

The influence of reperitonization on the induction of formation of intraperitoneal adhesions by a polypropylene mesh prosthesis. An experimental study in rats.

The correction of groin hernias has increasingly been performed by transperitoneal videolaparoscopic methods, using a polypropylene mesh. This could lead to an increased incidence of adhesion formation. The incidence of adhesions induced by mesh placement and the influence of reperitonization was observed on 40 male adult Wistar rats. Adhesions were significantly more common on the groups in which the prosthesis was placed (59% vs 95%; p = 0.01), as well as on the groups in which reperitonization was performed (58% vs 100%; p = 0.03). The results suggest that polypropylene mesh placement and reperitonization are each independent factors in the induction of adhesions formation.

Can the IVIM model be used for renal perfusion imaging?

OBJECTIVE: Renal perfusion imaging may provide information about the hemodynamic significance of a renal artery stenosis and could improve noninvasive characterization when combined with angiography. It was proposed previously that diffusion sequences could provide useful perfusion indices based on the intravoxel incoherent motion (IVIM) model. Owing to motion artifacts, diffusion imaging has been restricted to relatively immobile organs like the brain. With the availability of single-shot echo-planar imaging (EPI) our purpose was to evaluate the IVIM model in renal perfusion. METHODS AND MATERIAL: Eight volunteers underwent diffusion-sensitive magnetic resonance (MR) imaging of the kidneys using a spin echo (SE) EPI sequence. The diffusion coefficients determined by a linear regression analysis and fits to the IVIM function were calculated. RESULTS AND CONCLUSION: Our preliminary experience does not support the possibility of obtaining perfusion information using the IVIM model in the kidneys.

[The focus in neonatology]. / Le point en néonatologie.

During the last years, neonatology has greatly improved. In the last decade, mortality and morbidity have decreased: mortality from respiratory failure of prematurity has decreased from 22% to 12%, mortality of the very low birthweight infants under 1000 g fell from 56% to 35% and mortalities related to asphyxia have diminished from 21% to 12% and to malformations from 33% to 28%. Prematurity is now the first cause of neonatal mortality. During this period, the number of babies under 1000 g has increased 4-fold and the number of multiple births increased more than 2-fold from 3% to 7% of the live births of our hospital. Attitudes towards the premature infant have changed, especially towards the extremely small (called the micropremies). The number of disabled children has increased in parallel with the better survival of the very immature newborns who till recently were not resuscitated.