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different Patterns of Fluorescence in the Quantisation of anti-Leishmania Infantum Antibodies by Indirect Immuno-Fluorescent Antibody Test (IFAT) in the Serological diagnosis of Canine Leishmaniasis

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Open Journal of Veterinary Medicine 2017年第3期7卷 23-30页

作者： Pasquale Santoro Andrea Vellusi Università degli Studi di Napoli Federico II Dipartimento di Pediatria Napoli Italy diagnostica di Laboratorio (di.Lab.) S.r.l. Napoli Italy

Although Indirect Immuno-Fluorescent Antibody Test (IFAT), performed employing “in house” prepared antigen, is considered by several authors as the golden standard for the quantisation of anti-leishmania antibodies in dogs, there is a lack of papers reporting a description of the different patterns of fluorescence that can be observed. An incorrect identification of patterns of fluorescence may be an important source of bias in the interpretation of results. Previous papers report different criteria to define as “positive” a specific pattern of fluorescence, namely: membrane fluorescence, homogeneous fluorescence of the body, or homogeneous fluorescence of the body plus flagellum. In this paper, we report a detailed description of preparation of slides and of the patterns of fluorescence that can be obtained employing “in house” prepared antigen. At least six main patterns of fluorescence may be observed: 1): homogeneous cytoplasmatic green fluorescence;2): membrane pattern, in which the fluorescence is mainly localized along the entire perimeter of the parasites;3): coarse-speckled cytoplasmatic fluorescence;4): flagellar pattern, in which the fluorescence is localized exclusively onto the flagellum;5): punctiform pattern, in which the fluorescence is localized exclusively at the basis of the flagellum;6): nuclear pattern, in which only the nucleus of the parasite shows a homogeneous green fluorescent. The significance of each pattern is discussed.

关键词： Leishmaniasis Leishmania Infantum IFAT Dog diagnostics Fluorescence Pattern

The Serological diagnosis of Canine Leishmaniasis: Commercial ELISA or in House IFAT?

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Open Journal of Veterinary Medicine 2015年第2期5卷 30-34页

作者： Pasquale Santoro Andrea Vellusi Università degli Studi di Napoli Federico II - dipartimento di pediatria Napoli Italy diagnostica di Laboratorio (di.Lab.) S.r.l. Napoli Italy

The levels of anti-Leishmania infantum antibodies are relevant with the diagnosis and the followup of infected dogs. Both ELISA (Enzyme-Linked Immunosorbent Assay) and IFAT (Indirect Immuno-Fluorescent Antibody Test) may be employed to estimate the antibody levels in dogs, but at present, the correlation between ELISA absorbance and IFAT titer has not been investigated. In this paper, we compared the optical densities obtained with a commercial ELISA, Leiscan?(Laboratorios Dr. Esteve S.A.), versus the titers obtained by in house IFAT. We measured ELISA absorbance of: 44 IFAT negative samples coming from an endemic area;10 negative samples coming from a non endemic area;29 samples with an IFAT titer between 1/40 and 1/80;10 samples with an IFAT titer of 1/160;9 samples with an IFAT titer of 1/320;10 samples with an IFAT titer of 1/640;10 samples with an IFAT titer ≥ of 1/1280. Results show that: a) there is a poor correlation between IFAT titer and ELISA absorbance;b) Leiscan? is not able to distinguish between IFAT negative samples and IFAT titers up to 1:160;c) IFAT negative samples from an endemic or non endemic area show different ELISA absorbance;d) the performance of the kit may be improved by the use of a more appropriate cut-off.

关键词： ELISA IFAT Leishmania infantum

diagnosis of Laron syndrome using monoplex-polymerase chain reaction technology with a whole-genome amplification template: A case report

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World Journal of Clinical Cases 2019年第23期7卷 4029-4035页

作者： Adina Neumann Miguel Angel Alcantara-Ortigoza Ariadna Gonzalez-del Angel Felipe Camargo-diaz Esther Lopez-Bayghen Laboratorio de Investigacion y diagnostico Molecular Instituto de Infertilidad y Genetica Mexico SCINGENESMexico City 05320Mexico Instituto Nacional de pediatria Torre de InvestigacionMexico City 04530Mexico Laboratorio de Biologia Molecular Departamento de Genetica HumanaInstituto Nacional de PediatriaMexico City 04530Mexico Departamento de Toxicologia Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional(CINVESTAV-IPN)Mexico City 07360Mexico

BACKGROUND Laron syndrome(LS)is an autosomal recessive hereditary condition affecting only 1/1000000 *** cause is associated with mutations in the growth hormone(GH)receptor(GHR),leading to GH *** patients typically present with severe growth retardation,obesity,and abnormal sexual ***,LS diagnosis is performed ***,we assessed the efficiency of Pre-implantation Genetic Testing(PGT)coupled with monoplex-polymerase chain reaction(PCR)technology for detecting this monogenic disease in embryos from a couple confirmed as LS heterozygous carriers CASE SUMMARY The couple LS-carriers were confirmed by the presence of a first child born with *** couple underwent a standard in vitro fertilization(IVF)*** was collected from trophectoderm cells from day 5 *** genome amplification(WGA)was performed using a Sureplex DNA Amplification System and analyzed by PCR,targeting the deletion of the exons 5 and 6 in the GHR gene as well as PGT by Next-generation Sequencing(Illumina).Eleven embryos were collected and analyzed.27.3%were the wild type for GHR,45.5%were heterozygotes,and 18.2%homozygous *** embryo yielded no *** 2-embryos transfers were performed;2 normal homozygous and four heterozygous carriers were selected for *** first two transfers were unsuccessful,whereas the final transfer with two heterozygous embryos resulted in clinical *** genomic composition of the fetus was verified,applying the same techniques using amniocytes,extracted after 21 wk of the ongoing *** fetus was confirmed as GHR deletion in exon 5-6,carrier.A non-affected baby was *** Here,we present a case demonstrating that using WGA as a template in addition to PCR targeting specific gene regions,exons 5 and 6 on the GHR gene,could identify LS carrier *** provides evidence that WGA and PCR serve as an excellent tool to detect this specific monogenic disease in IVF e

关键词： Growth hormone insensitivity Growth hormone receptor mutations Intragenic deletions Molecular diagnosis Embryo diagnosis Laron syndrome Case report

Occult hepatitis B virus infection among Mexican human immunodeficiency virus-1-infected patients

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World Journal of Gastroenterology 2014年第37期20卷 13530-13537页

作者： Ma Teresa Alvarez-Muoz Angelica Maldonado-Rodriguez Othon Rojas-Montes Rocio Torres-Ibarra Fernanda Gutierrez-Escolano Guillermo Vazquez-Rosales Alejandro Gomez Onofre Muoz Javier Torres Rosalia Lira Unidad de Investigacion Medica en Enfermedades Infecciosas y Parasitarias Hospital de PediatriaCentro Medico Nacional Siglo XXIInstituto Mexicano del Seguro Social Clinica de Hepatitis Hospital de Infectologia Centro Médico Nacional La Raza Hospital General Regional No.72 IMSS Unidad de Investigacion Medica en Enfermedades Infecciosas y Parasitarias Hospital de PediatriaCentro Medico Nacional Siglo XXIInstituto Mexi-cano del Seguro Social Hospital Infantil de Mexico "Federico Gomez"

AIM: To determine the frequency of occult hepatitis B infection (OHBI) in a group of human immunodeficiency virus (HIV)-1+/ hepatitis B surface antigen negative (HBsAg)- patients from Mexico.

关键词： Hepatitis B virus Occult hepatitis B virus infection Human immunodeficiency virus Hepatitis B surface antigen negative Risk factors Molecular diagnostics

A Pilot Experience in the training of healthcare professionals to face the childhood obesity epidemic through family therapeutic education

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Open Journal of Pediatrics 2013年第2期3卷 165-176页

作者： Tanas Rita Mazza Federica Caggese Guido Rossato Elisa Donach Martin Marcolongo Renzo dipartimento di Medicina Clinica e Sperimentale Azienda Ospedaliera-Università Padua Italy dipartimento Emergenza Azienda Ospedaliero-Universitaria Ferrara Italy Struttura Interaziendale di Formazione e Progetti Internazionali Azienda Ospedaliero Universitaria Padua Italy University School of Medicine New York USA UO pediatria Ospedaliera Azienda Ospedaliero-Universitaria Ferrara Italy

Background: Obesity is a high-cost social disease, the management of which, usually assigned to general practice, is less supported by evidence-based medicine. Most general practitioners feel unprepared to face the problem. Objectives: In 2000, at the Pediatric Department of Ferrara, we started a Family Group Therapeutic Education Program for the treatment of childhood obesity. More than 300 families have attended the program. Following our positive results, we created a professional training course on therapeutic education, addressed to healthcare personnel involved in the treatment of childhood obesity. The purpose of the present study is to discover if it is possible to promote the development of a shared therapeutic education strategy for obesity, involving primary care physiccians/pediatricians, specialists and dietitians. Methods: The integrated professional training course consists of a four-day seminar along with an on-line course for distance learning. The assessment of the project takes into account the following criteria: a multidimensional questionnaire (a pre- and post-test which explores: knowledge, motivation, self-efficacy, methodology, communication, modeling, etc.), an appreciation questionnaire with responses collected over the course of 2 years. Moreover, the nine participants of our 2008 training course began to use the new therapeutic tools in their practice and six of them sent us the anthropometric measurements of patients whom they have treated over the past two years using this approach allows us to present outcomes in terms of implementation of this therapeutic education program in everyday professional practice. Results and Conclusions: The course was attended by 15 healthcare professionals: ten in 2008 and five in 2009, from different Italian centers. Results seem to indicate a great interest and appreciation by the participants. Results based on BMI z-score reduction of the children cared for by our trainees seem to support the efficacy

关键词： Pediatric Obesity Therapy Family-Based Treatment Therapeutic Patient Education Group Therapy

1b型糖原贮积症中基因型和表现型间的相关性:一项多中心研究和文献回顾

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世界核心医学期刊文摘（儿科学分册） 2006年第1期2卷 19-20页

作者： Melis D. Fulceri R. Parenti G. 贺莉 dipartimento di pediatria Universit Federico II Via Sergio Pansini 5 80131 Napoli Italy

We studied the genotype/phenotype correlation in a cohort of glycogen storage disease type (GSD) 1b patients. A total of 25 GSD1b patients, 13 females and 12 males, age range: 4.3-28.4 years, mean:14.6±6.8 years; median: 15 years, repre senting the entire case load of Italian GSD1b patients, were enrolled in the stu dy. Molecular analysis of the glucose 6-phosphate translocase (G6PT1) gene was performed in all patients. We analysed the presence of a correlation among both the clinical features associated with GSD1b (neutropenia, frequency of admission to the hospital for severe infections) and the presence of systemic complicatio ns (liver adenomas, nephropathy, bone mineral density defect, polycystic ovaries , short stature, in-flammatory bowel disease) and the mutations detected in each pati ent. Nine patients were homozygous or compound heterozygous for mutations causin g stop codons. In particular, three patients were homozygous for the same mutati on (400X); of these patients, one showed chronic neutropenia with severe and fre quent infections and severe inflammatory bowel disease, another patient cyclic n eutropenia associated with rare bacterial infections and mild bowel involvement and the last one normal neutrophil count. Two patients were homozygous for the m utation 128X; one of these patients did not show neutropenia, whereas the other one had severe neutropenia needing frequent hospital admission and was under gra nulocyte-colony stimulating factor treatment. In three patients no mutations we re detected. Conclusion:no correlation was found between individual mutations an d the presence of neutropenia, bacterial infections and systemic complications. These results suggest that different genes and proteins modulate neutrophil diff erentiation, maturation and apoptosis and thus the severity and frequency of inf ections. The absence of detectable mutations in three patients could suggest tha t a second protein plays a role in microsomal phosphate transport

关键词：糖原贮积症文献回顾表现型粒细胞减少症身材矮小症磷酸转移酶多囊性卵巢分子学纯合子骨密

小儿肥胖相关性肝病时的代谢、激素、氧化和炎症因子

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世界核心医学期刊文摘（儿科学分册） 2005年第12期1卷 33-34页

作者： Mandato C. Lucariello S. Licenziati M.R. P. Vajro 郭战宏 dipartimento di pediatria Università di Napoli Federico IIVia S. Pansini 580131Napoli ItalyDr.

To examine the role of metabolic, hormonal, oxidative, and inflammatory factors in pediatric obesity-related liver disease. Study design: In 50 obese children (age 7 to 14 years) with (n = 20, group 1) or without (n = 30, group 2) hypertransaminasemia and ultrasonographic liver brightness, we studied insulin resistance (fasting glucose/insulin ratio [FGIR]- ) and serum levels of leptin, iron, transferrin, ferritin, C-reactive protein (CRP), white blood cell (WBC) count, tumor necrosis factor (TNF)- α , interleukin (IL)- 6, C282Y and H63D mutations, and erythrocytic glutathione peroxidase (GPX) activity. Results: FGIR (6.7 ± 4.1 vs 9.2 ± 5.2; P = .02), serum ferritin (88.8 ± 36.0 vs 39.9 ± 24.0 ng/mL; P = .0001), serum CRP (5.4 ± 6.0 vs 1.1 ± 1.6 mg/dL; P = 0.004), and GPX (8.4 ± 0.9 vs 5.0 ± 0.5 U/g Hb; P = .05) were significantly higher and more frequently deranged in group 1 than in group 2. FGIR, ferritin, and CRP values were simultaneously deranged in 41% of the group 1 patients and in none of the group 2 patients (P = .098). Serum leptin, iron, and transferrin, WBC, TNF- α , IL- 6, and C282Y and H63D mutations were similar in the 2 groups. Conclusions: Insulin resistance, oxidative stress, and low-grade systemic inflammatory status are implicated in pediatric obesity-related liver disease. These findings may be useful in planning pathophysiologically based therapeutic trials for hepatopathic obese children who are unable to follow hypocaloric diets.

关键词：炎症因子胰岛素抵抗过氧化酶血清铁蛋白空腹血糖肿瘤坏死因子肥胖儿童低热量饮食红细胞谷胱甘肽氧化应激

Prevention of Femoral Head Deformity after Ischemic Necrosis Using Ibandronate Acid and Growth Factor in Immature Pigs

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Surgical Science 2012年第4期3卷 194-199页

作者： Bibiana Dello Russo Eduardo Luis Baroni Nicolas Saravia Valeria Amelong Fabiana Lubiniecky Marcelo Asprea Gustavo Williams Susana Rodriguez J. P. Garrahan Hospital Nacional de pediatria Ciudad Autónoma de Buenos Aires Buenos Aires Argentina

Background: Femoral head deformity is the most severe sequela of ischemic necrosis in skeletally immature patients. Development of severe deformity shortens useful survival time of the joint due to the appearance of early degenerative changes. Preservation of the trabecular architecture through inhibition of osteoclastic bone resorption may minimize the development of the deformity in an animal model of ischemic necrosis of the femoral head. Aims: To determine if a highly potent antiabsorptive agent, ibandronate, would inhibit bone resorption during necrotic femoral head repair to avoid subsequent flattening and deformity, to determine if the use of platelet-rich plasma stimulates bone repair and neovascularization of the damaged femoral head, and to evaluate if the combination of both therapies can preserve the femoral head while stimulating new bone formation in an animal model of ischemic necrosis. Methods: Ischemic necrosis of the femoral head was induced by surgical ligature of the circumflex vessels in 10 Landrace pigs. The animals were divided into four different groups and were administered ibandronate acid, platelet-rich plasma, or both. The contralateral, untreated femoral heads with surgical ligature of the circumflex vessels served as the control group. All animals were killed three months after surgery and the femoral head was evaluated both radiographically and histologically. The femur length was measured on radiographs and compared among the ***: Final femoral length was significantly longer in the group treated with a combination of both therapies (platelet-rich plasma-ibandronate acid) compared to the others groups, with a significant difference between groups. The histological findings showed increased osteoblastic activity and thickened trabiculae, a higher rate of neovascularization, and focal hyperplasia greater bone resorption and neovascularization. Only slight changes (femoral length) were observed in the animals that received pla

关键词： Perthe’s disease Avascular Necrosis Ibandronate Acid Platelet-Rich Plasma

Low Efficiency of the Commonly Prescribed Drugs against Klebsiella pneumonia, Escherichia coli and Acinetobacter Species as the Causative Agents of Blood Stream Infection in Malabo, Equatorial Guinea

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Advances in Microbiology 2016年第3期6卷 162-168页

作者： Aleksey Shatalov Yaari Shilo Rizeq Nakhash Dennis Zhdanov Department of Microbiology La Paz Medical Center Malabo Equatorial Guinea Intern Medicine Department La Paz Medical Center Malabo Equatorial Guinea Department of Neonatology and pediatria La Paz Medical Center Malabo Equatorial Guinea Intensive Care unit Department La Paz Medical Center Malabo Equatorial Guinea

The prevalence of multi drug resistant gram-negative bacteria to commonly first line drugs in blood is a serious problem in Equatorial Guinea and other world. This is the first study describing antibiotic resistance analysis of blood stream infection in Equatorial Guinea. Our study presents alarming rate of inefficiency of the most commonly prescribed drugs to treatment Klebsiella pneumoniae, Escherichia coli and Acinetobacter species isolates as the most frequency etiologic agents in blood stream infection. Out of 1849 blood culture the bacterial etiological agents were isolated from 196 (10.6%) samples. E. coli (n = 22), K. pneumonia (n = 39) and Acinetobacter (n = 17) represent 71.6% of all gram negative bacterial isolates. Almost all isolates of K. pneumonia and Acinetobacter sp. (92.1% and 100%, respectively) and about 50% of E. coli strains possessed extended-spectrum β-lactamase activity. Alarming level of multi drug resistant gram negative strains was observed. E. coli and K. pneumonia and Acinetobacter isolates demonstrated low sensitivity to all commonly prescribed drugs such as Ampicillin, Trimethoprim/Sulfamethoxazole, Doxycycline, Gentamycin Amoxicicline/Clavulanic Acid, Cefuroxime, Ciprofloxacine. It is especially worth noting the low efficiency of third generation cephalosporins (Cefrtiaxon) against Acinetobacter and Klebsiella with their resistance rate of 94.7% and 100% respectively. Moreover, the alarming level of low sensitivity to Piperacilin/Tazobactam of K. pneumonia (22%) and Acinetobacter (29.4%) was been found. The 17.6% of Acinetobacter isolated was carbomenem resistant. Just Imipenem and Amikacin were the most sensitive drug against these bacterial strains.

关键词： Bacteremia Multi Drug Resistance ESBL Gram Negative Rods