Gene Polymorphisms of TNF-α<Superscript>−308</Superscript>, IL-10<Superscript>−1082</Superscript>, IL-6<Superscript>−174</Superscript>, and IL-1Ra<Superscript>VNTR</Superscript> Related to Susceptibility and Severity of Rheumatic Heart Disease

Rheumatic heart disease (RHD) is an inflammatory disease of the heart tissues caused by interactive immune, genetic, and environmental factors. The objective of this study is to test for the association of polymorphisms related to cytokine genes with susceptibility and severity of RHD among affected children from the Nile Delta region of Egypt. The study included 50 children with chronic RHD (29 males and 21 females), with a mean age of 12.2 years, in addition to 98 healthy unrelated controls. Cases were further classified on the basis of echocardiographic findings into those with only mitral valve disease (MVD) or multivalvular lesions (MVLs) and also as mild, moderate, or severe valve lesions. For all cases and controls, DNA was extracted and amplified using polymerase chain reaction with sequence-specific primers for detection of single nucleotide polymorphisms (SNPs) in the promoter regions of cytokine genes tumor necrosis factor (TNF)-alpha(-308 )G/A, interleukin (IL)-10(-1082 )G/A, and IL-6(-174 )G/C as well as a variable number of tandem repeats (VNTRs) in intron 2 of the IL-1Ra gene. All cases showed a significantly higher frequency of homozygous genotypes of TNF-alpha(-308 )A/A [odds ratio (OR) = 5.7, p < 0.001], IL-10(-1082) A/A (OR = 3.1, p < 0.05), IL-10(-1082) G/G (OR = 5.2, p < 0.05), and IL-1Ra A1/A1 (OR = 2.2, p < 0.05). Cases with MVD showed higher frequencies of genotypes TNF-alpha(-308 )A/A, G/G; IL-10(-1082) G/G; and IL-1Ra(VNTR) A1/A1 (p < 0.05). Cases with MVL showed a significantly higher frequency of homozygous A/A genotype of both TNF-alpha(-308 )(OR = 10.6, p < 0.05) and IL-10(-1082) (OR = 5.2, p < 0.05). The same was observed for cases with severe valve lesions. On the other hand, all studied groups showed significantly lower frequency of heterozygous genotypes of TNF-alpha(-308 )G/A, IL-10(-1082) G/A, and IL-1Ra(VNTR) A1/A2. No significant difference was found regarding the frequency of IL-6(-174 )G/C polymorphisms in total cases or subgroups compared to controls (p > 0.05). Predisposition to RHD is influenced by genetic factors including cytokine gene polymorphisms, with possible susceptibility to severe disease with multivalvular affection among cases with composite polymorphism (TNF-alpha(-308 )A/A and IL-10(-1082) A/A) and (TNF-alpha(-308 )A/A and IL-10(-1082) G/G).     

Comparison of the Occlutech<Superscript>®</Superscript> Figulla<Superscript>®</Superscript> Septal Occluder and Amplatzer<Superscript>®</Superscript> Septal Occluder for Atrial Septal Defect Device Closure

The Occlutech^® Figulla^® septal occluder (OFSO) is a later-generation double-disk device with few reports of its success rates and complications compared with the Amplatzer^® septal occluder (ASO), which is the worldwide standard device in percutaneous atrial septal defect (ASD) closure. We recruited and compared the results in 149 patients (76.5 % female) who underwent ASD device closure in our center between January 2003 and June 2012. The patients ranged in age from 2.3 to 77.2 years. There were no statistically significant differences between the two groups regarding patient baseline characteristics and procedure variables. The success rate using either device was excellent (ASO 94.4 % and OFSO 97.4 %; p = 0.43). Although the diameter of the ASD and the pulmonary arterial pressure in the OFSO group were slightly higher than in the ASO group, the median fluoroscopic time in the OFSO group was significantly shorter (ASO 13.7 min; OFSO 9.0 min; p < 0.001). The overall median follow-up time was 3.6 years (interquartile range 2.1–9.0 years). There were no significant differences between the major and minor complications when comparing the two devices. Both devices were safe and effective for percutaneous ASD closures. The OFSO had the benefit of a shorter fluoroscopic time.     

Selective deficiency of naïve CD4<Superscript>+</Superscript> T-lymphocytes in a child with congenital lymphoedema

We describe the first known case of congenital lymphoedema associated with selective deficit of naïve CD4⁺ T-lymphocytes. A high proportion of naïve CD4⁺ T-lymphocytes was found in the ascitic fluid, supporting the hypothesis of extra-vascular sequestration of these cells into lymphoedematous tissue.Abbreviation ICL idiopathic CD4⁺ lymphocytopenia     

The use of Surgisis<Superscript><Emphasis Type="Bold">®</Emphasis></Superscript> for abdominal wall reconstruction in the separation of omphalopagus conjoined twins

Abdominal wall reconstruction in omphalopagus twins poses a difficult reconstructive challenge, as separation often results in a large abdominal wall defect. A number of options are available for closure, including tissue flaps, expanders and patches made of foreign material. Surgisis is a new biodegradable small intestine scaffolding substrate that permits tissue in-growth and results in a permanent durable scar. We describe its use in abdominal wall reconstruction after separation of a set of conjoined twins. A set of omphalopagus conjoined twins shared liver and abdominal wall. After separation at 6 months of age, Twin A's abdomen could be closed primarily, but Twin B could not. A 4-ply Surgisis mesh was used in the upper abdominal closure, and a skin flap was created, to completely cover the patch. Both twins survived the operation. A small portion of the skin flap over the Surgisis broke down, healing by secondary intention. In follow up of over 18 months post procedure, there have been no wound infections and the abdominal wall is intact with no evidence of a hernia. Surgisis can be successfully used for the reconstruction of complex abdominal wall defects in the pediatric patient, including reconstruction after separation of conjoined twins.     

Matriderm<Superscript>®</Superscript> 1 mm versus Integra<Superscript>®</Superscript> Single Layer 1.3 mm for one-step closure of full thickness skin defects: a comparative experimental study in rats

Purpose

Dermal templates, such as Matriderm^® and Integra^®, are widely used in plastic and reconstructive surgery, often as two-step procedures. A recent development is the application of thin dermal templates covered with split thickness skin grafts in one-step procedures. In this experimental study, we compare the two thin matrices Matriderm^® 1 mm and Integra^® Single Layer in a one-step procedure with particular focus on neodermis formation.

Methods

Matriderm^® 1 mm and Integra^® Dermal Regeneration Template—Single Layer (1.3 mm) were compared in a rat model. In three groups of five animals each, a full thickness wound was covered with (a) Matriderm^® 1 mm and neonatal rat epidermis, (b) Integra^® Single Layer and neonatal rat epidermis, or, (c) neonatal rat epidermis only (control). Histological sections 2 weeks post transplantation were analyzed with regard to take of template and epidermis, neodermal thickness, collagen deposition, vascularization, and inflammatory response.

Results

Take of both templates was complete in all animals. The Matriderm^®-based neodermis was thinner but showed a higher cell density than the Integra^®-based neodermis. The other parameters were similar in both matrices.

Conclusion

The two templates demonstrate a comparable biological behavior early after transplantation. The only difference was found regarding neodermal thickness, probably resulting from faster degradation of Matriderm^®. These preliminary data suggest that both dermal templates appear similarly suitable for transplantation in a one-step procedure.

     

Ranitidine-enhanced <Superscript>99m </Superscript>technetium pertechnetate imaging in children improves the sensitivity of identifying heterotopic gastric mucosa in Meckel’s diverticulum

Meckels diverticulum is the most common congenital gastrointestinal anomaly. ^99m Technetium pertechnetate imaging (Meckels scan) is the best noninvasive method used to diagnose this condition when heterotopic gastric mucosa (HGM) is present. Although cimetidine enhancement has been shown to improve sensitivity of the Meckels scan, ranitidine enhancement has also been advocated; however, this recommendation is based on unpublished data. Thirty-seven children with confirmed Meckels diverticulum were reviewed retrospectively. Of eight children with HGM in the Meckels diverticulum who presented with profuse rectal bleeding and underwent the conventional Meckels scan, three of them (37.5%) had a false negative study. Ranitidine, when administered either intravenously or orally for 24 h prior to the Meckels scan, enhanced the sensitivity of this test to 87.5% in our patient cohort.     

Disseminated Bacillus Calmette-Guérin lymphadenitis in a patient with gp91<Emphasis Type="Italic">phox</Emphasis><Superscript>−</Superscript> chronic granulomatous disease 25 years after vaccination

A boy developed ipsilateral axillary lymphadenitis after Bacillus Calmette-Guérin (BCG) inoculation at the age of 5 months. Subsequently, he was diagnosed with X-linked chronic granulomatous disease (CGD) by the nitroblue tetrazolium assay when he was 4 years old. Body computerized tomography (CT) performed at the age of 25 years showed enlarged lymph nodes in the left periclavicular and axillary regions, and was confirmed by gallium scintigraphy. Mycobacterial culture, smear, and polymerase chain reaction (PCR) of the sputum and gastric fluid were negative. Whole-blood IFN-γ assay was negative as well. Mycobacterium bovis BCG was isolated from the lymph node biopsy by PCR amplification and culture. No mutation of the IFN-γ receptor 1 could be identified. In conclusion, CGD can be the underlying condition for BCG-itis; whole-blood IFN-γ assay might be useful in differentiating BCG infection and tuberculosis in CGD patients; BCG vaccination is contraindicated in X-linked CGD.     

Incorporation of Integra<Superscript>®</Superscript> in tissue defects: a pilot study in the rat model

Integra has been shown to be very useful in accelerating the growth of neodermis. It has found extensive use in case of burns as a primary dressing immediately after a burn, after release of contractures and following scar revision. It has been used to achieve cover after the debridement of extensive infective processes involving the skin. Encouraged by these results we have assessed the application of Integra to augment and/or patch defects of the urinary bladder, diaphragm and the abdominal wall in the rat model. This was a pilot study and involved the incorporation of Integra in the diaphragm, the urinary bladder (extramucosal) and the muscle layer of the abdominal wall. Eight adult Wistar rats were given general anaesthesia and Integra was implanted with absorbable sutures at the sites mentioned. The omentum was hitched to the collagen matrix surface to revascularise the graft. The silicone was left in situ. The operative period was covered with antibiotics. The anaesthesia was then reversed. Postoperatively the rats were given analgesia and feeds started immediately. The rats were sacrificed after 3 weeks. The abdominal cavity was examined for adhesions. The Integra implant along with adjacent tissue was harvested and examined histologically. There were no visible intra-abdominal adhesions. The histology revealed good degree of neovascularisation and fibrosis in and adjacent to the implant. This was comparable to the changes seen in the skin. This pilot study has shown that implanting Integra invokes a similar response in deeper tissues and it can develop neovascularisation from the omentum. Hence, this could find some application in treating congenital conditions such as diaphragmatic hernias, abdominal wall defects and for bladders requiring augmentation. Our initial results are quite encouraging and we feel that this field should be further explored.     

A unique service in UK delivering Plastibell<Superscript>®</Superscript> circumcision: review of 9-year results

Muslim infants undergo circumcision for religious reasons and Bradford has a high Muslim population. The National Health Service in UK does not provide religious circumcision, so in 1996 a nurse-delivered circumcision service led by consultant urologists was set up at a no-profit and cost-only basis. Plastibell circumcision was offered to all infants between 6 and 14 weeks old and performed under local anaesthesia. Information leaflets and videotapes about the procedure were available to parents prior to the procedure. A three monthly audit of the service was undertaken. Between July 1996 and June 2005 (9 years) 1,129 circumcisions were performed. The common complications were problems with the ring (3.6%) and bleeding (3%). Overall, there was 96% satisfaction rate among the service users. The Plastibell technique for circumcision is a simple method and can be safely performed by trained nurses with acceptable complication rates.     

Δ<Superscript>1</Superscript>-pyrroline-5-carboxylate synthase deficiency: neurodegeneration,cataracts and connective tissue manifestations combined with hyperammonaemia and reduced ornithine,citrulline, arginine and proline

¹-pyrroline-5-carboxylate synthase (P5CS) catalyses the reduction of glutamate to ¹-pyrroline-5-carboxylate, a critical step in the biosynthesis of proline, ornithine and arginine. Recently, we reported a newly recognised inborn error due to deficiency of P5CS in two sibs, one presenting at birth with hypotonia, dysmorphic signs, pes planus and clonic seizures. Both developed progressive neurodegeneration and peripheral neuropathy, joint laxity, skin hyperelasticity and bilateral subcapsular cataracts. Their metabolic phenotype includes mild hyperammonaemia, hypo-ornithinaemia, hypocitrullinaemia, hypo-argininaemia and hypoprolinaemia. Incorporation of ³H-proline into protein was deficient in fibroblasts incubated with ³H-glutamate. Both patients are homozygous for the missense mutation R84Q in P5CS. Here, we describe the clinical phenotype of the sibs in detail and show that a relative deficiency of urea cycle intermediates (ornithine, citrulline and arginine) during fasting periods results in a paradoxical hyperammonaemia. Furthermore, we show the results of ornithine loading tests and indirect enzyme studies corroborating the biological significance of the defect in P5CS in vivo. Conclusion:The metabolic phenotype of ¹-pyrroline-5-carboxylate synthase deficiency is easily missed. The combination of low levels of ornithine, citrulline, arginine and proline plus a tendency to hyperammonaemia or one of the above together with a clinical phenotype of neurodegeneration with peripheral neuropathy and/or cataracts and connective tissue manifestations should suggest this disorder. Early recognition would allow a therapeutic trial with citrulline and proline.     

Mechanism of Rho-kinase-mediated Ca<Superscript>2+</Superscript>-independent contraction in aganglionic smooth muscle in a rat model of Hirschsprung’s disease

Purpose  

Lack of ganglion cells is the main cause of bowel movement disorder in Hirschsprung’s disease. Because smooth muscle is the primary organ, the properties of intestinal smooth muscle need to be investigated. We therefore investigated the reactivity of the contractile system and the mechanism of contraction in aganglionic intestinal smooth muscle.     

Successful Use of Recombinant Factor VIIa (NovoSeven<Superscript>®</Superscript>) During Cardiac Surgery in a Pediatric Patient with Congenital Factor XI Deficiency

We report our experience with the use of recombinant activated factor VII (rFVIIa) during cardiac surgery in a 4.5-year-old boy with severe congenital FXI deficiency and a congenital heart disease. After weaning the patient from cardiopulmonary bypass, the first intravenous dose of rFVIIa (90 μg/kg) was administered. This same dosage was repeated eight more times, at 2- to 4-hour intervals postoperatively. There was no bleeding during and after surgery. rFVIIa treatment may be used successfully in children with severe FXI deficiency in major operations such as open heart surgery.     

过敏性紫癜患儿血清IFN-α IL-2IL-4 TNF-α水平的变化

目的 检测过敏性紫癜患儿血清ＩＦＮ－α，ＩＬ－２，ＩＬ－４及ＴＮＦ－α水平的变化，探讨细胞因子在过敏性紫癜发病机制中的作用。方法 采用双抗体夹心法ＥＬＩＳＡ检测上述细胞因子。结果 过敏性紫癜患儿急性期血清中ＩＦＮ－α、ＩＬ－４和ＴＮＦ－α水平均较正常儿高（Ｐ＜０．０５＝，而ＩＬ－２水平无明显差别（Ｐ＜０．０５＝；无肾脏功能改变患儿血清ＩＦＮ－α，ＩＬ－２与正常儿无明显差别，有肾脏功能改变患儿血清ＩＦＮ－α水平较正常儿明显升高，而ＩＬ－２水平则显著降低（Ｐ＜ ０．０５）。结论 过敏性紫癜患儿血清中存在细胞因子紊乱，并可能参与过敏性紫癜发病和肾脏损伤。     

Association of <Emphasis Type="Italic">IL-10</Emphasis> Gene Polymorphism (−819C &gt; T, −592C &gt; A and −1082G &gt; A) with Preterm Birth

Objective

To analyze the association of IL-10 gene and its polymorphisms with preterm birth (PTB).

Methods

Five hundred and fifty nine women with term birth and 559 with preterm birth were recruited from Lucknow, India. Genetic association analysis was conducted between cases and controls. Subjects recruited as cases were women (aged between 18–40 y) with singleton delivery before 37 wk of gestation and controls were with delivery after or on 37 wk. The genotyping was performed for rs1800871, rs1800872 and rs1800896 for assessing the allelic distribution, haplotypic association and linkage disequilibrium analysis. IL-10mRNA levels were evaluated by real time quantitative polymerase chain reaction (PCR) method.

Results

The risk of PTB was found significant in women carrying IL-10 (?1082) GA genotype [OR=1.72(1.7–2.5), p=0.006]. The haplotypic analysis of studied polymorphisms for rs1800871, rs1800872 and rs1800896 depicted the association of ATA (p=0.02) and ATC (p=0.01) haplotypes with PTB. The IL-10 mRNA levels were significantly lower in cases (p=0.05).

Conclusions

IL-10 marks a protective impact in the inflammatory pathway of PTB.

     

支气管哮喘模型大鼠IL-4 IL-10 IFN-γ的检测

目的　哮喘的发病机理尚不完全清楚 ,近年来 ,细胞因子在哮喘中的作用引起人们的关注。本文旨在探讨支气管哮喘的免疫学发病机理 ,研究细胞因子IL 4、IL 10、IFN γ与哮喘的关系。方法　建立大鼠支气管哮喘模型。将Wistar大鼠随机分为两组 :正常对照组 (n =11)及哮喘组 (n =11) ,以卵白蛋白 (OVA)腹腔注射致敏大鼠 ,两周后雾化吸入OVA发敏 ;发敏后杀鼠 ,取脾 ,制备单个核细胞悬液 ,体外培养 ,并以OVA刺激 ,取不同时间培养上清 ,应用ELISA双抗体夹心法测定其中IL 4、IL 10和IFN γ的水平。结果　支气管哮喘组动物单个核细胞培养上清中IL 4的水平明显高于正常对照组 (6 4 .5 6± 5 .83vs 2 4 .6 6± 3.6 8pg/ml,P <0 .0 5 ) ,IL 10水平明显低于正常对照组 (34.13± 0 .85vs 85 .12± 6 .13,P <0 .0 5 ) ,IFN γ水平明显低于正常对照组 (3.87± 0 .4 8vs14 .5 1± 1.32 pg/ml,P <0 .0 5 )。结论　细胞因子IL 4、IL 10和IFN γ可能参与了哮喘发病的病理生理过程。     

A novel <Emphasis Type="Italic">SOX10</Emphasis> mutation in a patient with PCWH who developed hypoxic–ischemic encephalopathy after <Emphasis Type="Italic">E. coli</Emphasis> sepsis

We describe a male infant with a novel SOX10 mutation and a severe course of PCWH—a special phenotype of Shah-Waardenburg syndrome involving peripheral demyelinating neuropathy, central dysmyelinating leukodystrophy, Waardenburg syndrome, and Hirschsprung’s disease. The patient had severe hypoplastic hypoganglionosis of the small and total colonic intestine together with peripheral and central dysmyelination. The patient was completely dependent on parenteral nutrition. We identified a novel frameshift mutation, p.Asp293GlyfsX10, in the SOX10 gene of this patient. The mutation would encode a protein that lacked the transactivation domain and resulted in the largest duplication described to date. At the age of 20 months, the boy presented with a severe complication with a translocation of Escherichia coli and developed sepsis leading to severe hypoxic–ischemic encephalopathy with persistent vegetative state (PVS). The boy died at the age of 24 months. Conclusion: Septic encephalopathy with hypoxic–ischemic encephalopathy can be a serious complication in severe sepsis. It is unknown to what extent the mutant SOX10 protein influenced the degree of brain injury—for example central nervous system susceptibility to hypoxia—during sepsis, which may explain the severe encephalopathy with clinical signs of PVS the boy developed.     

Severe combined immunodeficiency caused by a splicing abnormality of the <Emphasis Type="Italic"> CD3δ </Emphasis>gene

CD3 deficiency is a recently identified rare form of severe combined immunodeficiency. We analysed the CD3 gene in a Japanese family with severe combined immunodeficiency. The patients lacked T-cells with normal numbers of B-cells and natural killer cells in peripheral blood. We found a novel homozygous mutation in the splicing acceptor site of intron 2 (IVS2–2AG) in these patients. Analysis of patients mononuclear cells revealed the CD3 splicing abnormality. Chest X-ray films and computed tomography revealed small sized thymuses in these patients. Conclusion:The CD3 gene should be analysed in patients with severe combined immunodeficiency lacking T-cells with normal B- and natural killer cells irrespective of the thymus size.