A study of contralateral persistent processus vaginalis in laparoscopic hernia repair in children.

PURPOSE: The introduction of laparoscopy for hernia repair permits intra-abdominal observation of a hernia and contralateral persistent processus vaginalis (CPPV). The current study's aim was to investigate the diameter of opening of an inguinal hernia and CPPV in patients with unilateral inguinal hernia, and to evaluate their correlation with age. METHODS: From September 2012 to August 2017, 569 pediatric patients underwent laparoscopic repair of unilateral inguinal hernia. We retrospectively evaluated the size of the hernia and CPPV by measuring the diameter of opening. Pearson correlation analysis and linear-by-linear association were used in the statistical analysis. RESULTS: The median age at operation was 32.4 months (range 0.2-219 months). CPPV was observed in 330 patients (58.0%), and its incidence was significantly higher in patients with left inguinal hernias than in those with right inguinal hernias (62.8 versus 54.0%, p < 0.001). The mean diameter of opening for the hernias was significantly larger than that for CPPV (11.2 ± 3.1 vs. 6.1 ± 2.5 mm, p < 0.001). The incidence of CPPV gradually decreased from 77.2% in infants to 46.6% in the oldest age group (≥ 6 years) (p trend < 0.001). The diameter of the opening of a hernia was not correlated with increasing age, and the diameter of the opening of a CPPV was not correlated with increasing age, as well. CONCLUSION: The diameters of a hernia and CPPV were identified in the current study, and the diameter was not correlated with increasing age. The incidence of CPPV was more common in patients with left inguinal hernias than in those with right inguinal hernias, and it gradually decreased with increasing age.

Immunization of mice with live-attenuated late liver stage-arresting Plasmodium yoelii parasites generates protective antibody responses to preerythrocytic stages of malaria.

Understanding protective immunity to malaria is essential for the design of an effective vaccine to prevent the large number of infections and deaths caused by this parasitic disease. To date, whole-parasite immunization with attenuated parasites is the most effective method to confer sterile protection against malaria infection in clinical trials. Mouse model studies have highlighted the essential role that CD8(+) T cells play in protection against preerythrocytic stages of malaria; however, there is mounting evidence that antibodies are also important in these stages. Here, we show that experimental immunization of mice with Plasmodium yoelii fabb/f(-) (Pyfabb/f(-)), a genetically attenuated rodent malaria parasite that arrests late in the liver stage, induced functional antibodies that inhibited hepatocyte invasion in vitro and reduced liver-stage burden in vivo. These antibodies were sufficient to induce sterile protection from challenge by P. yoelii sporozoites in the absence of T cells in 50% of mice when sporozoites were administered by mosquito bite but not when they were administered by intravenous injection. Moreover, among mice challenged by mosquito bite, a higher proportion of BALB/c mice than C57BL/6 mice developed sterile protection (62.5% and 37.5%, respectively). Analysis of the antibody isotypes induced by immunization with Pyfabb/f(-) showed that, overall, BALB/c mice developed an IgG1-biased response, whereas C57BL/6 mice developed an IgG2b/c-biased response. Our data demonstrate for the first time that antibodies induced by experimental immunization of mice with a genetically attenuated rodent parasite play a protective role during the preerythrocytic stages of malaria. Furthermore, they highlight the importance of considering both the route of challenge and the genetic background of the mouse strains used when interpreting vaccine efficacy studies in animal models of malaria infection.

Short-lived effector CD8 T cells induced by genetically attenuated malaria parasite vaccination express CD11c.

Vaccination with a single dose of genetically attenuated malaria parasites can induce sterile protection against sporozoite challenge in the rodent Plasmodium yoelii model. Protection is dependent on CD8(+) T cells, involves perforin and gamma interferon (IFN-γ), and is correlated with the expansion of effector memory CD8(+) T cells in the liver. Here, we have further characterized vaccine-induced changes in the CD8(+) T cell phenotype and demonstrated significant upregulation of CD11c on CD3(+) CD8b(+) T cells in the liver, spleen, and peripheral blood. CD11c(+) CD8(+) T cells are predominantly CD11a(hi) CD44(hi) CD62L(-), indicative of antigen-experienced effector cells. Following in vitro restimulation with malaria-infected hepatocytes, CD11c(+) CD8(+) T cells expressed inflammatory cytokines and cytotoxicity markers, including IFN-γ, tumor necrosis factor alpha (TNF-α), interleukin-2 (IL-2), perforin, and CD107a. CD11c(-) CD8(+) T cells, on the other hand, expressed negligible amounts of all inflammatory cytokines and cytotoxicity markers tested, indicating that CD11c marks multifunctional effector CD8(+) T cells. Coculture of CD11c(+), but not CD11c(-), CD8(+) T cells with sporozoite-infected primary hepatocytes significantly inhibited liver-stage parasite development. Tetramer staining for the immunodominant circumsporozoite protein (CSP)-specific CD8(+) T cell epitope demonstrated that approximately two-thirds of CSP-specific cells expressed CD11c at the peak of the CD11c(+) CD8(+) T cell response, but CD11c expression was lost as the CD8(+) T cells entered the memory phase. Further analyses showed that CD11c(+) CD8(+) T cells are primarily KLRG1(+) CD127(-) terminal effectors, whereas all KLRG1(-) CD127(+) memory precursor effector cells are CD11c(-) CD8(+) T cells. Together, these results suggest that CD11c marks a subset of highly inflammatory, short-lived, antigen-specific effector cells, which may play an important role in eliminating infected hepatocytes.

Microtubule- and dynein-mediated movement of Orientia tsutsugamushi to the microtubule organizing center.

The host cell microfilaments and microtubules (MTs) are known to play a critical role in the life cycles of several pathogenic intracellular microbes by providing for successful invasion and promoting movement of the pathogen once inside the host cell cytoplasm. Orientia tsutsugamushi, an obligate intracellular bacterium, enters host cells by induced phagocytosis, escapes to the cytosol, and then replicates in the cytosol. ECV304 cells infected with O. tsutsugamushi revealed the colocalization of the MT organizing center (MTOC) and cytosolic orientiae by indirect immunofluorescence assay. Using immunofluorescence microscopy in the presence and absence of MT-depolymerizing agents (colchicine and nocodazole), it was shown that the cytosolic oriential movement was mediated by MTs. By transfection study (overexpression of dynamitin [also called p50], which is known to associate with dynein-dependent movement), the movement of O. tsutsugamushi to the MTOC was also mediated by dynein, the minus-end-directed MT-related motor. Although the significance of this movement in the life cycle of O. tsutsugamushi was not proven, we propose that the cytosolic O. tsutsugamushi bacteria use MTs and dyneins to propel themselves from the cell periphery to the MTOC.

Cellular invasion of Orientia tsutsugamushi requires initial interaction with cell surface heparan sulfate.

Role of transmembrane heparan sulfate proteoglycans on invasion of Orientia tsutsugamushi into host cells was investigated. Pretreatment with heparan sulfate and heparin inhibited the infection of O. tsutsugamushi for L cell, mouse fibroblast, whereas other glycosaminoglycans had little effect. These same treatments were also shown to reduce the infection in a dose-dependent manner, and enzymatic treatment of cells with heparitinase, but not chondroitinase ABC, inhibited the infection. In addition, mutant cell lines of Chinese hamster ovarian cell defective in heparan sulfate synthesis but not chondrotin sulfate synthesis and defective in all glycosaminoglycan synthesis showed marked reduction in susceptibility to infection by O. tsutsugamushi. Also mutant cell lines, which express heparan sulfate proteoglycans at low level, showed intermediate level of infectivity. Finally O. tsutsugamushi bind to(35)S-labelled heparin. Collectively, these findings provide strong evidence that heparan sulfate proteoglycans contribute to the attachment of O. tsutsugamushi to the cells.

Observations of the liquid-crystal analog of the abrikosov phase.

Freeze-fracture transmission electron micrographs of the smectic A(*) phase confirm the twist grain boundary model of Renn and Lubensky. The fracture surface has an undulating structure with a 0.5-micrometer helical pitch parallel to 4.1-nanometer smectic layers. The layers are disrupted by a lattice of screw dislocations oriented normal to the helical axis. Optical diffraction shows that rotation of smectic blocks occurs in discrete steps of about 17 degrees ; hence, the screw dislocations are 14 to 15 nanometers apart and the grain boundaries are 24 nanometers apart. These observations show that the SmA(*) phase is the liquid-crystal analog of the Abrikosov phase in superconductors.

Polymorphism of a long-chain cycloparaffin (CH2)120.

The polymorphism of a long-chain cycloparaffin (CH2)120 and chain packing in its crystals were discussed on the basis of some results obtained mainly by transmission electron microscopy. Monoclinic and orthorhombic single crystals of (CH2)120 were isothermally grown together from a dilute solution in p-xylene. Lozenge-shaped orthorhombic single crystals were more frequently observed than lath-shaped monoclinic ones. The basal surfaces of orthorhombic and monoclinic single crystal platelets were decorated with vapor-deposited polyethylene [PE]. Orthorhombic single crystals of (CH2)120 with the (110) twin boundary and monoclinic ones with the (100) twin boundary were also observed. Rod-like edge-on crystals of (CH2)120 were grown from a dilute p-xylene solution onto the (001) surface of alkali halides. The crystal system of the (CH2)120 edge-on crystals depended on the kind of substrate. The monoclinic crystal was grown on NaCl, the orthorhombic one on KBr and KCl. The monoclinic form of (CH2)120 edge-on crystal was transformed to the orthorhombic one by annealing on NaCl. In both monoclinic and orthorhombic edge-on crystals, the molecular plane determined by two zigzag stems in a molecule of (CH2)120 was parallel to the substrate surface and the molecular axis (crystallographic c-axis) was perpendicular to the longer side of the rod-like edge-on crystals. The sub-cell dimensions of the stem chains in both forms of (CH2)120 crystals were very similar to those of monoclinic and orthorhombic PE crystals, respectively.