A purse-string suture at the level of internal inguinal ring, taking only the peritoneum leaving the distal sac: is it enough for inguinal hernia in pediatric patients?

PURPOSE: Although laparoscopic surgery in children has expanded in recent years. Laparoscopic hernia repair in children is still debatable. We aimed to summarize and describe our results of laparoscopic inguinal hernia repair and techniques among children. METHODS: Between March 2011 and April 2013, 98 children (67 male, 31 female) underwent laparoscopic inguinal hernia repair at the department of surgery. The clinical outcomes were collected retrospectively. RESULTS: The mean follow-up period was 22.6 months. Twelve patients were ex-premature infants and a contralateral patent processus vaginalis (PPV) was present in 37 of the 91 unilateral inguinal hernia patients. There were two postoperative complications (transient hydrocele, umbilical port site infection). The mean operative time was 46 min. Recurrence, metachronous hernia and testicular atrophy were not observed during the follow-up period. CONCLUSIONS: Our preliminary experiences suggest that the laparoscopic purse-string suture of internal inguinal opening of hernia sac could be a safe, effective, and reliable alternative for management of pediatric inguinal hernia.

Sphingosine-1-phosphate signaling in human submandibular cells.

Sphingosine-1-phosphate (S1P) is a significant lipid messenger modulating many physiological responses. S1P plays a critical role in autoimmune disease and is suggested to be involved in Sjögren's syndrome pathology. However, the mechanism of S1P signaling in salivary glands is unclear. Here we studied the effects of S1P on normal human submandibular gland cells. S1P increased levels of the intracellular Ca(2+) concentration ([Ca(2+)](i)), which was inhibited by pre-treatment with U73122 or 2-aminoethoxydiphenyl borate (2-APB). Pre-treated S1P did not inhibit subsequent carbachol-induced [Ca(2+)](i) increase, which suggests that S1P and muscarinic signaling are independent of each other. S1P1, S1P2, and S1P3 receptors SphK1 and SphK2 were commonly expressed in human salivary gland cells. S1P, but not carbachol, induces the expression of interleukin-6 and Fas. Our results suggest that S1P triggers Ca(2+) signaling and the apoptotic pathway in normal submandibular gland cells, which suggests in turn that S1P affects the progression of Sjögren's syndrome.

Dopamine neuron glutamate cotransmission: frequency-dependent modulation in the mesoventromedial projection.

Mesoventromedial dopamine neurons projecting from the medial ventral tegmental area to the ventromedial shell of the nucleus accumbens play a role in attributing incentive salience to environmental stimuli that predict important events, and appear to be particularly sensitive to the effects of psychostimulant drugs. Despite the observation that these dopamine neurons make up almost the entire complement of neurons in the projection, stimulating their cell bodies evokes a fast glutamatergic response in accumbens neurons. This is apparently due to dopamine neuron glutamate cotransmission, suggested by the extensive coexpression of vesicular glutamate transporter 2 (VGLUT2) in the neurons. To examine the interplay between the dopamine and glutamate signals, we used acute quasi-horizontal brain slices made from DAT-YFP mice in which the intact mesoventromedial projection can be visualized. Under current clamp, when dopamine neurons were stimulated repeatedly, dopamine neuron glutamate transmission showed dopamine-mediated facilitation, solely at higher, burst-firing frequencies. Facilitation was diminished under voltage clamp and flipped to inhibition by intracellular Cs(+) or GDPbetaS, indicating that it was mediated postsynaptically. Postsynaptic facilitation was D1 mediated, required activation of NMDA receptors and closure of voltage gated K(+)-channels. When postsynaptic facilitation was blocked, D2-mediated presynaptic inhibition became apparent. These counterbalanced pre- and postsynaptic actions determine the frequency dependence of dopamine modulation; at lower firing frequencies dopamine modulation is not apparent, while at burst firing frequency postsynaptic facilitation dominates and dopamine becomes facilitatory. Dopamine neuron glutamate cotransmission may play an important role in encoding the incentive salience value of conditioned stimuli that activate goal-directed behaviors, and may be an important subtract for enduring drug-seeking behaviors.

Comparison of three warming devices for the prevention of core hypothermia and post-anaesthesia shivering.

The efficacy of forced air warming with a surgical access blanket in preventing a decrease in core temperature during anaesthesia and post-anaesthesia shivering (PAS) was compared with two widely used interventions comprising forced air warming combined with an upper body blanket, and a circulating water mattress, in a prospective, randomized double-blind study. A total of 90 patients undergoing total abdominal hysterectomy were studied, 30 in each group. Core temperature was measured 15, 30, 45, 60, 90 and 120 min after induction of anaesthesia. PAS was evaluated every 5 min after emergence from anaesthesia over a period of 1 h. Core temperature fell in all three groups compared with the baseline, but forced air warming using a surgical access blanket was more effective than the other warming methods in ameliorating the temperature decrease. The surgical access blanket was also superior to the circulating water mattress in reducing PAS.

EMLA cream versus dorsal penile nerve block for postcircumcision analgesia in children.

We conducted a prospective, randomized, double-blinded trial comparing preoperative application of EMLA cream and sodium chloride solution dorsal penile block (n = 31) with placebo cream and bupivacaine dorsal penile nerve block (n = 32) for postcircumcision analgesia. Pain was assessed using modified Children's Hospital of Eastern Ontario Pain Scale and the duration of block by the time to requirement of first dose of postoperative analgesic. There was no difference in Children's Hospital of Eastern Ontario Pain Scale between the two groups, but bupivacaine dorsal penile nerve block resulted in longer analgesia (P = 0.003). There were no local or systemic complications related to either technique, and there was a very small incidence of vomiting. We conclude that preoperative application of EMLA cream is an effective and simple method to produce postcircumcision analgesia with a very small incidence of adverse effects.

Application of cDNA microarray technique to detection of gene expression in host cells infected with viruses.

cDNA microarray technique was used to monitor changes in mRNA levels in cells after Hantaan virus (HTNV) infection. The values of the ratio of medians for HTNV and Japanese encephalitis virus (JEV) at the early stage of infection were compared and found similar, suggesting that the same or similar genes are associated with the early events of infection with either virus. The reproducibility of values of the "ratio of medians" for HTNV was examined. We found that applying cluster analysis to the gene expression data groups efficiently together genes with the same function. Therefore, in analyzing the effects of viral infection on host cells by the cDNA microarray technique, clustering data appear to be necessary for gaining biological meaning from a dump of gene expression profiles obtained from virus-infected cells.

Chip electrochromatography of polycyclic aromatic hydrocarbons on an acrylate-based UV-initiated porous polymer monolith.

The first rigorous evaluation of a UV-initiated porous polymer monolith (PPM) as a stationary phase for chip electrochromatography (ChEC) is described. All channels in an offset T-injector-design-chip (25-microm deep by 50-microm wide channels) were filled by capillary action with an acrylate-based PPM precursor solution and polymerized in situ using 365 nm light for several minutes. Photodefinability of the monolith cast in the channels during the polymerization process was also demonstrated by masking off the injection arms during photoinitiation. The chromatographic performance of this chip was compared with that of chips completely filled with monolith. The detection window was photodefined after polymerization using the detection laser (257 nm doubled argon ion laser) to depolymerize the detection window. A successful ChEC separation of 10 out of 13 polycyclic aromatic hydrocarbons (PAH) was performed with on-column, off-packing laser-induced fluorescence detection at 257 nm. Van Deemter plots for early-, middle-, and late-eluting compounds showed the minimum plate height to be 5 microm. The average number of theoretical plates per meter for the PAH was 200,000. Several factors contributed to irreproducible results. Oxygen was observed to dynamically quench the fluorescence of the sample over time. Improved sealing of the reservoirs solved this problem. A within-chip variability in the retention time of 2-10% RSD was observed. These results demonstrate the feasibility and reliability of the PPM as a solid reversed-phase for electroosmotic flow-driven chip-based chromatography in microscale total analysis systems.

Profiling of impurities in heroin by capillary electrochromatography and laser-induced fluorescence detection.

Capillary electrochromatography (CEC) with laser-induced fluorescence (LIF) detection was investigated for the analysis of acidic and neutral impurities in heroin. The phenanthrene-like heroin impurities exhibit high native fluorescence when excited with a doubled argon ion laser (operating at 257 nm). The limit of detection for acetylthebaol is 66 pg ml(-1). CEC-LIF analysis of heroin samples of different geographical origin gave distinguishable peak-enriched chromatograms. A sulfonic acid C12 polymer monolith column provided similar resolving power to a 1.5 mm non-porous ODS column for the isocratic analysis of a refined heroin sample. Analysis of a crude heroin sample via a multi-step gradient CEC resolved a significantly higher number of peaks than gradient high-performance liquid chromatography or micellar electrokinetic capillary chromatography.

Conduct-as-cast polymer monoliths as separation media for capillary electrochromatography.

We have developed porous polymer monoliths (PPMs) that are versatile and robust reversed-phase chromatography media. The PPMs are cast-to-shape, UV-cured polymers that form uniform packings within pretreated glass capillaries and fused-silica chips. No applied pressure is ever needed to flush the PPMs since they support electroosmotic flow as cast. Such characteristics make the PPMs useful for chip-based devices. Our results show efficiencies greater than or equal to 150,000 plates/m for both capillary and chip-based separations of polycyclic aromatic hydrocarbons. By changing the monomers, the hydrophobicity of the polymers, and the direction of the electroosmotic flow can be altered without degrading chromatographic performance. We describe here the development of these acrylate-based materials along with both physical and chromatographic characterization.

The use of intravenous atropine after a saline infusion in the prevention of spinal anesthesia-induced hypotension in elderly patients.

UNLABELLED: We investigated the efficacy of IV atropine for preventing spinal anesthesia-induced hypotension in elderly patients. Seventy-five patients undergoing transurethral prostate or bladder surgery were randomized to receive either placebo (n = 25), atropine 5 microg/kg (small-dose atropine, n = 25) or atropine 10 microg/kg (large-dose atropine, n = 25) after the induction of spinal anesthesia. All the patients received an IV infusion of 10 mL/kg 0.9% normal saline over 10 min before the induction of anesthesia. The systolic blood pressure decreased in all three groups after spinal anesthesia. There was a significant increase in the mean heart rate in both atropine groups as compared to the placebo group (placebo group: 78 bpm, 95% confidence interval [CI]: 76.6-78.5; small-dose atropine group: 86 bpm, 95% CI 83.9-88.8; large-dose atropine group: 97 bpm, 95% CI 94.5-100.3; P: = 0.001). There was a significant decrease in the incidence of hypotension in patients who received atropine (placebo group: 76%, small-dose atropine group: 52%, large-dose atropine group: 40%, P: = 0.03). The mean dose of ephedrine required was significantly decreased in the atropine groups (placebo group: 12.2 mg [SD= 10.5], small-dose atropine group: 7.4 mg [SD= 10.0], large-dose atropine group: 5.4 mg [SD= 8.7 mg], P: = 0.048). The total amount of IV fluid and number of patients requiring metaraminol in addition to 30 mg of ephedrine were not significantly different among the three groups. Significant side effects, such as confusion, ST segment changes or angina were not detected in any of the patients. We conclude that IV atropine may be a useful supplement to the existing methods in preventing hypotension induced by spinal anesthesia. IMPLICATIONS: IV atropine increases heart rate in a dose-dependent manner in elderly patients undergoing spinal anesthesia. It reduces the incidence of hypotension and the dose of ephedrine required. Small-dose atropine may be a useful supplement in preventing spinal anesthesia-induced hypotension in elderly patients.

Ultrahigh-resolution plastic graded-index fused image plates.

Fiber-optic imaging systems such as the medical endoscope, the boroscope, the fused-image faceplate, and the image conduit are now made from glass step-index (SI) fibers, and the image resolution of the SI fiber-optic imaging systems is limited to ~5 microm. Ultrahigh-resolution fiber-optic fused-image plates with fiber diameter sizes of 5 and 2.8 microm were fabricated with plastic graded-index (GRIN) fibers. The measured image resolutions of the 5-microm SI and GRIN-based guides were comparable, and the resolution of the plastic GRIN image guides improved as the fiber diameter was reduced from 5 to 2.8 microm.

Antibody reaction of human anti-Toxoplasma gondii positive and negative sera with Neospora caninum antigens.

Anti-Neospora caninum antibody was detected in anti-Toxoplasma gondii positive and negative human sera by ELISA, western blot and immunofluorescence assay (IFA). Twelve cases out of 172 (6.7%) Toxoplasma-positive sera cross-reacted with both T. gondii and N. caninum antigens, and one out of 110 Toxoplasma-negative sera reacted with N. caninum antigen by ELISA. By western blot, all 12 sera reacted with T. gondii antigens with various banding patterns but specifically at 30 kDa (SAG1) and 22 kDa (SAG2) bands. With N. caninum antigen, the number of reactive bands was reduced, however a 43 kDa band reacted in three cases in Toxoplasma-positive sera in addition to one in Toxoplasma-negative control sera. All sera of the Toxoplasma-positive group labeled surface membrane of T. gondii, but reacted differently with N. caninum. Fluorescence was detected in surface membrane, subcellular organelles, or both in N. caninum. And one case in the Toxoplasma-negative group also reacted with N. caninum strongly in subcellular organelles. This suggested that the antibody against N. caninum may be present in human sera although the positive rate was very low in this study. The possibility of human infection with N. caninum remains to be evaluated further.

Foodborne outbreaks of human toxoplasmosis.

Two outbreaks of acute toxoplasmosis involving 8 adult patients in Korea were linked to eating uncooked pork. In the first outbreak, 3 patients developed unilateral chorioretinitis within 3 months of eating a meal consisting of raw spleen and liver of a wild pig. In the second outbreak, 5 of 11 soldiers who ate a meal consisting of raw liver of a domestic pig developed lymphadenopathy. All 8 patients had high levels of IgG Toxoplasma gondii antibodies (> or = 1:1024) in the Sabin-Feldman dye test, modified agglutination test incorporating mercaptoethanol, and latex agglutination test. T. gondii IgM antibodies persisted in these patients for several months. Most patients had a favorable response to anti-T. gondii chemotherapy with pyrimethamine and sulfanomides.

Analysis of antigenic domain of GST fused major surface protein (p30) fragments of Toxoplasma gondii.

Antigenic domain of major surface protein (p30) of Toxoplasma gondii was analyzed after polymerase chain reaction (PCR) of its gene fragments. Hydrophilic or hydrophobic moiety of amino acid sequences were expressed as glutathione S-transferase (GST) fusion proteins. Fragments of p30 gene were as follows: T37, total p30 open reading frame (ORF); S28, total ORF excluding N-terminal signal sequence and C-terminal hydrophobic sequence: A19, N-terminal 2/3 parts of S28; P19, C-terminal 2/3 of S28; X9, N-terminal 1/3 part of S28; Y10, middle 1/3 of S28; and Z9, C-terminal 1/3 of S28, respectively. Primer of each fragment was synthesized to include clamp sequence of EcoR I restriction site. PCR amplified DNA was inserted into GST (26 kDa) expression vector, pGEX-4T-1 to transform into Escherichia coli (JM105 strain). GST fusion proteins were expressed with IPTG induction as 63, 54, 45, 45, 35, 36, and 35 kDa proteins measured by SDS-PAGE. Each fusion protein was confirmed with GST detection kit. Western blot analysis with the serum of a toxoplasmosis patient revealed antigenicity in proteins expressed by T37, S28, and A19 but not those by P18, X9, Y10, and Z9. Antigenicity of p30 seems to be located either in N-terminal 1/3 part in the presence of middle 1/3 part or in the oligopeptides between margins of the first and second 1/3 parts.

Inhibition of Con A-induced lymphocyte proliferation by peritoneal exudate of Toxoplasma gondii-infected mice.

The presence of biological response modifiers (BRM)-like effect was confirmed in peritoneal exudate (PE) of Toxoplasma gondii-infected ICR mice which inhibited Concanavalin A (Con A)-induced peritoneal lymphocyte (PL) proliferation. During 5 days of PL incubation with 10 micrograms/ml Con A with or without PE, 3H-thymidine uptake was measured for the last 24 hrs. Compared to uninduced control, PL proliferated by 7.3-fold with Con A induction. When PE of infected mice was added, PL proliferation was inhibited by 74.0 +/- 11.9% whereas inhibition by PE of normal mice was 16.4 +/- 8.3%. Inhibitory effect of PE increased exponentially from 3 days up to 4-5 days of survival after the infection. Inhibitory activity of PE was decreased concentration-dependently. Also the inhibition was diminished when the PE was treated with heat of 95 degrees C for 10 min or precipitated with 10% trichloracetic acid (TCA). In SDS-PAGE of PE, many minor bands appeared newly. Heat-labile protein molecule in PE exerted inhibitory activity to Con A-induced lymphocyte proliferation.

Lymphadenitis in experimental murine toxoplasmosis induced by intramuscular injection of tachyzoites.

When tachyzoites (RH strain) of Toxoplasma gondii are injected intramuscularly, experimental mice survive up to 7 days, 1-2 days longer than those infected intraperitoneally. We observed sequential histopathological changes in inguinal lymph nodes after intramuscular injection of tachyzoites to thighs of specific pathogen free (SPF) mice. Initial findings on 1 or 3 days after the injection were reactive germinal centers, distended sinuses and epithelioid cell clusters in cortical and paracortical regions. Later on 5 days after the injection, however, effacement of nodal structure with depletion of cells and focal necrosis were observed. Necrotizing lymphadenitis in the experimental murine toxoplasmosis suggests the causal relation between T. gondii infection and the human disease.

Culture of tissue-cyst forming strain of Toxoplasma gondii and the effect of cyclic AMP and pyrimidine salvage inhibitors.

An in vitro culturing to examine the cyst stage of Toxoplasma gondii (ME49 strain) was investigated using murine peritoneal macrophages, and we also examined the effect of cAMP or DHFR inhibitors on the growth of bradyzoites. For experiments ICR mice were injected i.p. with 1,500 brain cysts. At 1, 3, 5 and 7 days, peritoneal exudates were isolated and then adherent peritoneal macrophages were cultured for 1, 3, 5 and 10 days. Growth pattern of bradyzoites was measured by [3H]-uracil uptake assay and morphological pattern of pseudocysts formed in macrophages was observed with Giemsa stain. Mostly bradyzoites were observed in the macrophages extracted at 3 and 5 days post infection. After 3 days in vitro, a number of pseudocysts were formed in the macrophages and the size of pseudocysts was increased during further 5 and 10 days in vitro culture. cAMP stimulated the growth of bradyzoites when in vivo 3 and 5 days and then in vitro 5 and 10 days conditions were applied. In case of DHFR inhibitors, pyrimethamine produced a linearly decremental effect with a conc.-dependent mode but methotrexate was not effective against intracellular bradyzoites or pseudocysts in this system. It was suggested that cyst-forming strain of T. gondii (ME49 strain) could be maintained and cultivated in vitro by use of murine peritoneal macrophages. In vivo 3 and 5 days and then in vitro 5 and 10 days conditions appeared to be suitable for culturing of bradyzoites. cAMP and pyrimethamine had an effect of stimulation and inhibition on the growth of bradyzoite, respectively.

Inhibition of entry of Toxoplasma gondii into MDCK cells by fetal bovine serum.

We experienced the partial inhibition of entry of Toxoplasma gondii into MDCK cells when the FBS was depleted from media. MDCK cells and Toxoplasma (RH strain) were co-cultured, the penetration was inhibited up to 60-80% with concentration-dependence of FBS. Inhibitory effect was clear when the conc. of FBS was over 1% (v/v) with 50% inhibition conc. of 5%. When Toxoplasma was pre-incubated with FBS and then applied to MDCK cells, there were no inhibitory effect, but when FBS was added to Toxoplasma-MDCK co-culture, the time of adding was critical with rapid inhibition. And when FBS was further treated with heat (95 degrees C, 10 min), the inhibitory effect was decreased slightly in both raw and inactivated FBS. The FBS factor(s) might participate to neutralize secreted materials which enhancing penetration or intervene between receptor-ligand binding at the moment of entry through sterically rather than functionally.

Detection of antibodies in serum and cerebrospinal fluid to Toxoplasma gondii by indirect latex agglutination test and enzyme-linked immunosorbent assay.

Sensitivity of anti-Toxoplasma antibody (IgG) test by enzyme-linked immunosorbent assay (ELISA) was evaluated in comparison with indirect latex agglutination (ILA) using 2,016 paired human samples of serum and cerebrospinal fluid (CSF). The samples were collected from neurologic patients in Korea with mass lesions in central nervous system (CNS) as revealed by imaging diagnosis (CT/MRI). When the sera were screened for anti-Toxoplasma antibody by ILA, 76 cases(3.8%) were positive (1:32 or higher titers). In the paired samples of CSF, no positive reactions were observed. When ELISA was performed using PBS extract of Percoll purified tachyzoites as antigen, cut-off absorbance was determined as 0.40 for serum and 0.27 for CSF tests. The antibody positive rates by ELISA were 7.0% in serum and 5.6% in CSF. Of them, 40 cases (2.0%) showed positive reactions in both serum and CSF. The antibody positive rates were higher in groups older than 40 years. The rates were higher in male (4.7% by ILA, 8.3% by ELISA) than in female (2.2% by ILA, 5.0% by ELISA). The rates in CSF showed no such sex difference. ELISA showed twice higher positive rates when serum was tested, and was sensitive enough to detect specific antibodies in CSF. Etiologic relations between positive antibody tests and CNS lesions remained unknown.