Herpes simplex virus spreads rapidly in human foreskin, partly driven by chemokine-induced redistribution of Nectin-1 on keratinocytes.

HSV infects keratinocytes in the epidermis of skin via nectin-1. We established a human foreskin explant infection model to investigate HSV entry and spread. HSV1 entry could only be achieved by the topical application of virus via high density microarray projections (HD-MAPs) to the epidermis, which penetrated beyond one third of its thickness, simulating in vivo microtrauma. Rapid lateral spread of HSV1 to a mean of 13 keratinocytes wide occurred after 24 hours and free virus particles were observed between keratinocytes, consistent with an intercellular route of spread. Nectin-1 staining was markedly decreased in foci of infection in the epidermis and in the human keratinocyte HaCaT cell line. Nectin-1 was redistributed, at the protein level, in adjacent uninfected cells surrounding infection, inducible by CCL3, IL-8 (or CXCL8), and possibly CXCL10 and IL-6, thus facilitating spread. These findings provide the first insights into HSV1 entry and spread in human inner foreskin in situ.

Herpes Simplex Virus type 1 infects Langerhans cells and the novel epidermal dendritic cell, Epi-cDC2s, via different entry pathways.

Skin mononuclear phagocytes (MNPs) provide the first interactions of invading viruses with the immune system. In addition to Langerhans cells (LCs), we recently described a second epidermal MNP population, Epi-cDC2s, in human anogenital epidermis that is closely related to dermal conventional dendritic cells type 2 (cDC2) and can be preferentially infected by HIV. Here we show that in epidermal explants topically infected with herpes simplex virus (HSV-1), both LCs and Epi-cDC2s interact with HSV-1 particles and infected keratinocytes. Isolated Epi-cDC2s support higher levels of infection than LCs in vitro, inhibited by acyclovir, but both MNP subtypes express similar levels of the HSV entry receptors nectin-1 and HVEM, and show similar levels of initial uptake. Using inhibitors of endosomal acidification, actin and cholesterol, we found that HSV-1 utilises different entry pathways in each cell type. HSV-1 predominantly infects LCs, and monocyte-derived MNPs, via a pH-dependent pathway. In contrast, Epi-cDC2s are mainly infected via a pH-independent pathway which may contribute to the enhanced infection of Epi-cDC2s. Both cells underwent apoptosis suggesting that Epi-cDC2s may follow the same dermal migration and uptake by dermal MNPs that we have previously shown for LCs. Thus, we hypothesize that the uptake of HSV and infection of Epi-cDC2s will stimulate immune responses via a different pathway to LCs, which in future may help guide HSV vaccine development and adjuvant targeting.

Complexity of gastroschisis predicts outcome: epidemiology and experience in an Australian tertiary centre.

BACKGROUND: Gastroschisis is a congenital anomaly of the fetal abdominal wall, usually to the right side of umbilical insertion. It is often detected by routine antenatal ultrasound. Significant maternal and pediatric resources are utilised in the care of women and infants with gastroschisis. Increasing rates of gastroschisis worldwide have led institutions to review local data and investigate outcomes. A collaborative project was developed to review local epidemiology and investigate antenatal and neonatal factors influencing hospital length of stay (LOS) and total parental nutrition (TPN) in infants born with gastroschisis. METHODS: We performed a five-year review of infants born with gastroschisis (2011-2015) at a major Australian centre. Complex gastroschisis was defined as involvement of stenosis, atresia, ischemia, volvulus or perforation and closed or vanishing gastroschisis. We extracted data from files and databases at the two participating hospitals, a major maternal fetal medicine centre and the affiliated children's hospital. RESULTS: There were 56 infants antenatally diagnosed with gastroschisis with no terminations, one stillbirth (2%) and one infant with 'vanishing' gastroschisis. The mean maternal age was 23.9 years (range, 15-39 years). The mean gestation at delivery was 36 weeks (range, 25-39+ 3 weeks). Of the 55 neonates who received surgical management, 62% had primary closure. The median LOS was 33 (IQR, 23-45) days and the median duration of TPN was 26 (IQR, 17-36) days. Longer days on TPN (median 35 vs 16 days, P = 0.03) was associated with antenatal finding of multiple dilated bowel loops. Postnatal diagnosis of complex gastroschisis was made in 16% of cases and was associated with both longer LOS (median 89 vs 30 days, P = 0.003) and days on TPN (median 46 vs 21 days, P = 0.009). CONCLUSION: Complex gastroschisis was associated with greater days on TPN and LOS. We found no late-gestation stillbirths and a low overall rate of 1.8%, suggesting the risk for stillbirth associated with gastroschisis is lower than previously documented. This information may assist counselling families. Improved data collection worldwide may reveal causative factors and enable antenatal outcome predictors.

Relay of herpes simplex virus between Langerhans cells and dermal dendritic cells in human skin.

The mechanism by which immunity to Herpes Simplex Virus (HSV) is initiated is not completely defined. HSV initially infects mucosal epidermis prior to entering nerve endings. In mice, epidermal Langerhans cells (LCs) are the first dendritic cells (DCs) to encounter HSV, but it is CD103(+) dermal DCs that carry viral antigen to lymph nodes for antigen presentation, suggesting DC cross-talk in skin. In this study, we compared topically HSV-1 infected human foreskin explants with biopsies of initial human genital herpes lesions to show LCs are initially infected then emigrate into the dermis. Here, LCs bearing markers of maturation and apoptosis formed large cell clusters with BDCA3(+) dermal DCs (thought to be equivalent to murine CD103(+) dermal DCs) and DC-SIGN(+) DCs/macrophages. HSV-expressing LC fragments were observed inside the dermal DCs/macrophages and the BDCA3(+) dermal DCs had up-regulated a damaged cell uptake receptor CLEC9A. No other infected epidermal cells interacted with dermal DCs. Correspondingly, LCs isolated from human skin and infected with HSV-1 in vitro also underwent apoptosis and were taken up by similarly isolated BDCA3(+) dermal DCs and DC-SIGN(+) cells. Thus, we conclude a viral antigen relay takes place where HSV infected LCs undergo apoptosis and are taken up by dermal DCs for subsequent antigen presentation. This provides a rationale for targeting these cells with mucosal or perhaps intradermal HSV immunization.

Patterns of reflux in gastroesophageal reflux disease in pediatric population of New South Wales.

This study is to determine the association of ambulatory pH monitoring (24hr pH) with symptoms of gastroesophageal reflux and its other investigations. The clinical and epidemiological profiles of subjects referred for reflux disorders are also studied. Symptoms or group of symptoms, profiles and prior investigations of 1259 consecutive pediatric subjects (with 1332 24hr pH studies performed) referred for evaluation of reflux disorders between 1988 and 2012 were retrospectively studied. Chi-square or fisher exact test was used for hypothesis testing, student t-test for the comparison of means and the Wilcoxon rank-sum test for comparing medians of continuous variables. Gastroesophageal reflux disease (GERD), defined as reflux causing major symptoms and complications, was diagnosed in 57.5% subjects of the total sample. Forty-three percent were girls and 56.7% were boys. The most common age group was between 4 months and 2 years (51.2%). Vomiting (64.4%) and irritability (74%) were the most common symptoms with the neurological conditions (23.2%) being the most frequent underlying condition. The parameters used in 24hr pH were significantly higher in those diagnosed with GERD (P < 0.0001). The prevalence of GERD was found to be significantly higher when both gastrointestinal and respiratory symptoms were present (P = 0.008) at 66.4% than when compared with gastrointestinal (56.5%) and respiratory (52.2%) symptoms in isolation. Symptoms alone were not reliable in diagnosing GERD. Only 57.5% had GERD among patients referred for reflux disorders. 24hr pH is reliable and should be considered routine in reflux disorders, as it identifies patients with pathologic reflux and avoids a needless surgery.

Aganglionosis with the absence of hypertrophied nerve fibres predicts disease proximal to rectosigmoid colon.

PURPOSE: The gold standard for the diagnosis of Hirschsprung's disease (HSCR) is the pathologic evaluation of a rectal biopsy that demonstrates the absence of ganglion cells and nerve fibre hypertrophy. However, it has been frequently reported that hypertrophic nerves may not be present in some variants like long-segment HSCR, total colonic aganglionosis, premature and very young infants. The aim of this study was to determine this association. METHODS: We performed a retrospective review of the HSCR database at our tertiary care children's hospital from 2000 to 2013. In order to analyse the relationship between the diameter of the nerve fibres and the level of aganglionosis, we classified the patient sample into two groups-fibres ≤40 and >40 µm. The groups were statistically compared with P < 0.05 being significant. RESULTS: Rectal biopsies of 92 patients confirmed as HSCR with definitive operation performed at the same institution were reviewed. The mean nerve diameter was 50.1 µm (range 20-87.5 µm). Nerve fibre diameter ≤40 µm was predictive of transition zone above the sigmoid colon. A specificity of 77.3 % and a likelihood ratio of 2.03 supported this perception. No correlation was noted between nerve fibre diameter and gestational age at birth, birth weight or age at biopsy. CONCLUSION: The absence of nerve fibre hypertrophy in the presence of aganglionosis on rectal biopsy specimens is predictive of long-segment HSCR.

Technical tips and advancements in pediatric minimally invasive surgical training on porcine based simulations.

BACKGROUND: Minimal access techniques have transformed the way pediatric surgery is practiced. Due to various constraints, surgical residency programs have not been able to tutor adequate training skills in the routine setting. The advent of new technology and methods in minimally invasive surgery (MIS), has similarly contributed to the need for systematic skills' training in a safe, simulated environment. To enable the training of the proper technique among pediatric surgery trainees, we have advanced a porcine non-survival model for endoscopic surgery. MATERIALS AND METHODS: The technical advancements over the past 3 years and a subjective validation of the porcine model from 114 participating trainees using a standard questionnaire and a 5-point Likert scale have been described here. Mean attitude scores and analysis of variance (ANOVA) were used for statistical analysis of the data. RESULTS: Almost all trainees agreed or strongly agreed that the animal-based model was appropriate (98.35%) and also acknowledged that such workshops provided adequate practical experience before attempting on human subjects (96.6%). Mean attitude score for respondents was 19.08 (SD 3.4, range 4-20). Attitude scores showed no statistical association with years of experience or the level of seniority, indicating a positive attitude among all groups of respondents. CONCLUSIONS: Structured porcine-based MIS training should be an integral part of skill acquisition for pediatric surgery trainees and the experience gained can be transferred into clinical practice. We advocate that laparoscopic training should begin in a controlled workshop setting before procedures are attempted on human patients.

Low-grade fibromyxoid sarcoma, a deceptively benign tumor in a 5-year-old child.

Low-grade fibromyxoid sarcoma (LGFMS) is very uncommon in the pediatric population with only 20% of reported cases under the age of 18. The youngest reported case to date has been in a 4-year-old child. Lesions are usually slow growing and asymptomatic, and locations described in children have included paravertebral, thigh and intrathoracic. Although benign in appearance, these lesions can behave aggressively, with local recurrence and distant metastases primarily to lungs. These lesions can be resistant to the usual chemotherapy and radiotherapy with surgical resection being the treatment of choice. We report a case of a 5-year-old boy who presented with a mass in the left buttock.

Presentation of glaucoma in an urban tertiary care hospital in South America: legal blindness and prevalence.

PURPOSE: To determine disease severity in glaucoma patients who presented to a tertiary care service for the first time and to determine the prevalence of different types of glaucoma. METHODS: Retrospective analysis of data of patients referred to the Glaucoma Service at the Santa Casa de Misericórdia de São Paulo, Brazil in 2007. A complete chart review from 448 patients was done; data regarding age, gender, ethnicity, family history, duration of the disease, previous treatment, best corrected visual acuity, cup-to-disc ratio, intraocular pressure, diagnosis and treatment were collected. Glaucoma was diagnosed by the presence of typical optic disk abnormalities, disregarding IOP values. RESULTS: 52.3% of patients presented visual acuity less than or equal to 20/200 and 67.7% presented cup-to-disc ratio between 0.8 and 1.0 in the worse-seeing eye; 13.4% of patients were considered legally blind. Primary open angle glaucoma was the most prevalent form of glaucoma (54.2%, 95% CI: 48.5-59.5) and chronic angle closure glaucoma was the second most frequent (11.5%, 95% CI: 8.25-15.5). CONCLUSION: A considerable rate of patients (almost 60% in the better-seeing eye and 70% in the worse-seeing eye) presented with advanced glaucoma. Strategies that create conditions for early diagnosis are deemed necessary to reduce glaucoma-related blindness in the Brazilian population.

Hepatic derangement following N-Acetylcysteine enemas in an infant with cystic fibrosis.

We discuss an infant with MI secondary to cystic fibrosis, who was managed surgically by a double barrel ileostomy for mid - small bowel atresia and developed severe faecal impaction in the post - operative period. The faecal impaction was treated successfully with oral NAC and 0.2% NAC contrast enemas. The patient's liver function tests revealed a dramatic increase in transaminases and bilirubin contemporaneous with the administration of the enemas. The levels showed a spontaneous improvement after discontinuation. This is only the second reported case of hepatotoxicity secondary to NAC enemas in the literature. While our experience offers modest support for the use of NAC, its efficacy is not yet proven and paediatric surgeons using NAC in the enema form need to closely monitor liver function contemporaneous with this agent's administration and adjust their treatment accordingly.

Impact of minimally invasive surgery on the pediatric surgical profession.

We conducted a survey among pediatric surgeons to examine the impact of the advent of minimally invasive surgery (MIS) on the pediatric surgical profession with respect to job satisfaction and training challenges. An invitation to participate in a web-based questionnaire was sent out to 306 pediatric surgeons. Apart from demographic details and training recommendations, parameters relevant to job satisfaction, including patient interaction, peer pressure, ethical considerations, academic progress, ability to train residents, and financial remuneration, were studied. The response rate was 38.2%. Working in a unit performing MIS was identified by 71% of respondents as the most effective and feasible modality of training in MIS. Inability to get away from a busy practice was the most common reason cited for inability to acquire MIS training. The overall responses to the job satisfaction parameters showed a positive trend in the current MIS era for patient interaction, ethical considerations, academic progress, and training residents, with a negative trend for peer pressure and financial remuneration. The enthusiastic minimally invasive surgeons (EMIS) were defined as those having more than 5 years of MIS experience and also performing more than 10% of their work using MIS. Of the 113 responses analyzed, 67 belonged to the EMIS category. Those belonging to the EMIS group were less likely to feel inadequate in training their residents, in meeting the felt needs of the patients, or to complain about peer pressure. They were more likely to consider MIS to be as relevant and beneficial in children as in adults. Embracing MIS, as represented by the EMIS group, correlated with an overall greater job satisfaction.

An optimum animal model for neonatal thoracoscopy.

PURPOSE: This study attempted to define an optimum animal model for neonatal thoracoscopy. MATERIALS AND METHODS: Seven rabbits and three 5-7-kg piglets were subjected to thoracoscopy by three pediatric surgeons. The outcome for the successful completion of esophageal anastomosis and additional procedures, including diaphragmatic plication and lung biopsy, were noted. RESULTS: Whereas esophageal anastomosis could be accomplished in all piglets, it could be achieved in only 1 rabbit. Additional procedures were possible in 2 piglets and no rabbits. Anesthetic complications were seen in 4 rabbits and 1 piglet. CONCLUSION: Our findings suggest that the piglet may be the optimum animal model for replicating neonatal thoracoscopy. The fragility of the rabbit, coupled with a limited intrathoracic working space, limits its use for thoracoscopy.

What causes intrathoracic gastric dilatation in a late-presenting diaphragmatic hernia?

A 13-year-old previously asymptomatic patient presented with abdominal pain and vomiting. Chest radiogram revealed a left intrathoracic stomach. Laparotomy confirmed a dilated, hypertrophic stomach herniating through a left Bochdalek hernia. The patient recovered after a period of prolonged gastric paresis. While highlighting the importance of considering this condition in the differential diagnosis of a cystic lesion in the chest, this article also postulates a mechanism for the hypertrophic gastric dilatation in a late presentation of a congenital diaphragmatic hernia.

[Sensitivity of size I stimulus in achromatic automated perimetry for detection of glaucomatous visual field defects: a comparative analysis with short wavelength automated perimetry and standard automated perimetry (SITA)]. / Sensibilidade do estímulo de tamanho I na perimetria computadorizada acromática para a detecção de defeitos glaucomatosos no campo visual: análise comparativa com as perimetrias de ondas curtas e acromática convencional (SITA).

PURPOSE: To compare sensitivity and specificity of achromatic perimetry with size I stimulus (WW-I) with standard automated perimetry (SITA strategy) and blue on yellow perimetry (SWAP) for early diagnosis of glaucoma. METHODS: Seventy-three eyes of 73 suspects, glaucomatous and normal subjects underwent automated perimetry with SITA 24-2, WW-I and SWAP. After application of specific criteria of abnormality, sensitivity, specificity, and the area under ROC (Receiver Operating Characteristic) curve were calculated for each of the examinations. The areas under the ROC curve were compared pair-wisely with the univariable Z test and significance set at 5%. RESULTS: WW-I displayed the greatest sensitivity (100%) and the lowest specificity (75.7%). SWAP revealed less sensitivity (69.4%); specificity, however, was higher (89.2%). Pairwise comparison of ROC curves showed no difference. CONCLUSION: WW-I revealed good sensitivity for the detection of early glaucomatous visual field defects. This technique adds to the available armamentarium for early functional diagnosis of glaucomatous optic neuropathy.

[Clinical correlation between structural and functional assessment in glaucoma: Armaly cup to disk ratio and Brusini glaucoma staging system]. / Correlação clínica entre estrutura e função no glaucoma: classificação estrutural de Armaly e o sistema de estadiamento funcional de Brusini.

PURPOSE: To evaluate the correlation between Armaly's structural systems with Brusini's functional staging in glaucomatous patients. METHODS: Seventy-eight eyes of forty-two patients with primary open-angle glaucoma underwent visual field testing with the Humphrey Field Analyzer II (program SITA 24-2) and optic disc evaluation with the 78D Volk lens by two independent observers. Optic disc was stratified according to cup/disc ratio (C/D), from 0.1 to 1.0. Mean deviation (MD) and pattern standard deviation (PSD) values from each eye were plotted on Brusini's staging system to derive a value for glaucoma severity. Both cup/disc ratio and Brusini's values were plotted on a diagram to assess correlation (Spearman's rho correlation coefficient). Armaly's values were logged in a logarithmic figure and the data plotted on a diagram with Brusini's values for each patient. RESULTS: Armaly's and Brusini's systems dysplayed a poor correlation (rho=0.340). Both unlogged and logged, Armaly's versus Brusini's plots resulted in a linear and weak correlation. CONCLUSION: Despite the weak correlation, Armaly's structural system and Brusini's staging system provided non-independent measures for glaucoma evaluation.

[Variation of visual acuity in young patients with ectopia lentis submitted to surgery]. / Variação da acuidade visual em pacientes jovens com ectopia lentis submetidos à cirurgia.

PURPOSE: To assess the results as to visual acuity of two different surgical procedures for ectopia lentis. METHODS: Fifty-one eyes of 28 patients (16 males and 12 females, mean age 16.00 +/- 8.5) with simple (19 cases) or Marfan syndrome-associated (9 cases) ectopia lentis with different levels of subluxation underwent lens extraction with implantation of intraocular lenses (IOL) with scleral fixation (21 cases) or by lens extraction with implantation of the intraocular lenses in the capsular bag expanded by endocapsular ring (RING) (30 cases). Result analysis emphasized pre- and postoperative visual acuity during a six-month follow-up. RESULTS: Both techniques showed significant increase of postoperative visual acuity with and without correction, but it was better among the cases operated on by scleral fixation of the intraocular lenses. More than the used technique, the preoperative subluxation grades were crucial as to the results. CONCLUSION: The two surgical techniques for correction of simple or Marfan syndrome-associated ectopia lentis are safe and effective, resulting in significant recovery of visual acuity, although surgical results are completely dependent on the preoperative lens subluxation grades.

[Location and frequency of visual field defects as measured by SITA (Swedish Interactive Threshold Algorithm) strategy in primary open angle glaucoma]. / Localização e freqüência das alterações de campo visual no glaucoma primário de ângulo aberto na estratégia SITA (Swedish Interactive Threshold Algorithm).

PURPOSE: To evaluate the frequency, location and pattern of visual field defects as measured by SITA strategy in primary open angle glaucoma (POAG). METHODS: Six thousand and two-hundred charts were reviewed. One hundred and fifty two patients who fulfilled the inclusion criteria were included. They were stratified according to severity into early, moderate and severe. The following visual field defects were defined: nasal step, paracentral scotomata, arcuate defect, Seidel scotomata, temporal defect, general reduction of sensitivity, annular scotomata and central island. Each examination was divided into sectors corresponding to optic disc regions. Student's t test was used to compare differences in frequency. RESULTS: In the early glaucoma group, the nasal superior step was the most frequent visual field defect, followed by paracentral scotoma and inferior nasal step. General reduction of sensitivity was noted in 7.8%. Defects were more common in the superior hemifield (p<0.05). CONCLUSION: The superior nasal step and paracentral scotoma were the most commun visual field defects in early primary open angle glaucoma as evaluated by SITA strategy, and the superior hemifield comprises most of the defects. General reduction of sensitivity was an early and rare visual field defect.

[The role of frequency doubling perimetry in the diagnosis of glaucoma: screening in employees of a public hospital in an urban area of São Paulo]. / Importância da perimetria de dupla freqüência na detecção do glaucoma: rastreamento em funcionários de hospital público numa área urbana de São Paulo.

PURPOSE: To study the prevalence of glaucoma in employees of a public hospital in São Paulo and to analyze the role of frequency doubling perimetry along with non-contact tonometry and direct ophthalmoscopy, in the screening for glaucoma. METHODS: 612 employees of the Central Hospital of the "Santa Casa de São Paulo" were evaluated from October 15 to 20, 2000; 438 were women and 174 men, with mean age of 45.05+/-7.7 years (range, 35 to 81 years); 437 were white, 104 mulatto, 43 black and 28 Asian. All examinations were done by 10 eye specialists with experience in glaucoma. All employees had both eyes examined, by means of frequency doubling perimetry (FDT), ophthalmoscopy (FO), and non-contact tonometry (TNC). RESULTS: 159 (25.98%) subjects presented with at least one abnormal testing; 5 (3.14%) had high IOP (TNC+); 13 (8.17%) presented suspicious disk (FO+); 110 (69.18%) presented abnormal FDT (FDP+); 8 (5.03%) TNC+ and FO+; 10 (6.28%) had TNC+ and FDT+; 9 (5.66%) presented FO+ and FDT+; 4 (2.51%) had TNC+, FO+ and FDT+; 12 (1.96%) individuals were diagnosed with glaucoma, four of whom (0.65%), with normal pressure glaucoma. CONCLUSIONS: Frequency doubling perimetry was found to be an important step in the screening for glaucoma in the study. Of the 12 individuals that had the diagnosis of glaucoma, 5 (41.6%) would not have been diagnosed if they had not undergone frequency doubling perimetry.

Does the surgical approach change the need for a retrograde pyelogram prior to pyeloplasty?

OBJECTIVE: The opinion on the use of retrograde ureteropyelography (RUPG) prior to routine pyeloplasty for an ureteropelvic (UPJ) obstruction has been divided. This study analyses the efficacy of a preoperative RUPG and determines if a dorsal lumbotomy (DL) approach offers any advantage in this situation. METHODS: This is a retrospective analysis of application of RUPG prior to pyeloplasty in children with ages ranging from 42 days to 16.2 years who underwent surgery at the Children's Hospital at Westmead between 2009 and 2013. RESULTS: We identified a total of 95 children with isolated UPJ obstruction, with 59 (62.1%) boys and 36 (37.8%) girls. Overall, open pyeloplasties were performed in 89 (42 DL: 47 loin incision) and the rest (n = 6) laparoscopically. Preoperative RUPG was performed in 58 (61%) and it provided additional information in 11 (18.9%) patients for whom the surgical approach was modified. Hospital stay, operative time, and time to full diet were shorter with the DL approach (p < 0.05). CONCLUSIONS: The current study suggests that RUPG is avoidable if the approach for pyeloplasty is through the conventional loin incision. The short-term advantages might rationalize the use of RUPG if a DL incision is employed.