Stem cell therapy for Hirschprung disease

Boala Hirschsprung (HSCR) este o anomalie congenitală a colonului care rezultă din eșecul formării sistemului nervos enteric, care duce la un segment disfuncțional îngustat al colonului cu lungimi variabile și care necesită intervenție chirurgicală. Fiziopatologia de bază include un defect în migrarea, proliferarea și diferențierea celulelor crestei neurale, parțial explicate prin modificări genetice și epigenetice identificate. În ciuda ratei ridicate de succes a intervențiilor chirurgicale curative, acestea sunt asociate cu rezultate adverse semnificative, precum enterocolita, incontinența fecală și constipația cronică. În plus, unii pacienți suferă de variante letale extinse ale bolii, toate acestea justificând necesitatea unui tratament alternativ. În ultimii 5 ani, s-au înregistrat progrese considerabile în cercetarea terapiei bazate pe celule stem a HSCR. Cu toate acestea, multe probleme importante rămân nerezolvate. Această revizuire va oferi informații generale concise despre HSCR, va sublinia viitoarele abordări ale terapiei pe bază de celule stem a HSCR, va revizui publicațiile cheie recente, va discuta provocările tehnice și etice cu care se confruntă domeniul înainte de interpretarea clinică și va aborda aceste provocări propunând soluții și evaluând abordările existente pentru a progresa în continuare.Hirschsprung disease (HSCR) is a congenital anomaly of the colon that results from failure of enteric nervous system formation, leading to a constricted dysfunctional segment of the colon with variable lengths, and necessitating surgical intervention. The underlying pathophysiology includes a defect in neural crest cells migration, proliferation and differentiation, which are partially explained by identified genetic and epigenetic alterations. Despite the high success rate of the curative surgeries, they are associated with significant adverse outcomes such as enterocolitis, fecal soiling, and chronic constipation. In addition, some patients suffer from extensive lethal variants of the disease, all of which justify the need for an alternative cure. During the last 5 years, there has been considerable progress in HSCR stem cell-based therapy research. However, many major issues remain unsolved. This review will provide concise background information on HSCR, outline the future approaches of stem cell-based HSCR therapy, review recent key publications, discuss technical and ethical challenges the field faces prior to clinical translation, and tackle such challenges by proposing solutions and evaluating existing approaches to progress further

UXO Assessment on the Romanian Black Sea Coast

This paper aims to provide the reader with the results of the Unexploded Ordnance (UXO) survey of the defensive historical naval minefields launched by the Romanian and German Navies on the Romanian Black Sea coast, during the Second World War. This UXO survey was carried out between 2015-2018 by the Romanian Navy’s hydrographic ship “Commander Alexandru Cătuneanu” and Romanian Mine Warfare Data Center, using towed side-scan sonar technology and oceanographic observations. After explaining the materials and methodology, the results are presented and discussed: mosaics of the minefields, side-scan images of UXO contacts, side-scan images of the wrecks that were sunk in the minefields and some visible natural geological features of the seafloor. It was concluded that most of the objects discovered are sinkers, wreck debris or parts of chains, which does not represent a danger to navigation.

Echinococcus granulosus infection in two free-ranging Lumholtz’s tree-kangaroo (Dendrolagus lumholtzi) from the Atherton Tablelands, Queensland

Abstract: Infection with the larval stage of the cestode, Echinococcus granulosus sensu lato (s.l.), causes hydatid disease (hydatidosis) in a range of hosts, including macropods and other marsupials, cattle, and humans. Wild macropods are an important sylvatic reservoir for the life cycle of E. granulosus (s.l.) in Australia, and so provide a conduit for transmission of hydatid disease to domestic animals and humans. Two Lumholtz’s tree-kangaroos (Dendrolagus lumholtzi) from the Atherton Tablelands of Far North Queensland were recently found to have hydatid cysts in both liver and lung tissues. Tree-kangaroos may travel across the ground between patches of forest but are primarily arboreal leaf-eating macropods. The finding of hydatid cysts in an arboreal folivore may indicate that the area has a high level of contamination with eggs of E. granulosus (s.l.). This finding may be of significance to human health as well as indicating the need for further investigation into the prevalence of hydatid disease in domestic stock, wildlife and humans living in this rapidly urbanizing region

Surgery in postoperator alkaline desease

Clinica de Chirurgie Generală şi Esofagiană, Spitalul Clinic “Sfânta Maria” Bucureşti, România, Al XI-lea Congres al Asociației Chirurgilor „Nicolae Anestiadi” din Republica Moldova și cea de-a XXXIII-a Reuniune a Chirurgilor din Moldova „Iacomi-Răzeșu” 27-30 septembrie 2011Introducere: Se analizează retrospectiv experienţa clinicii pe 30 de ani (1981-2010) privind diversia duodenală totală Y-Roux (DDT) în tratamentul bolii alcaline de reflux postoperator(BARP). Materiale şi metodă, rezultate: Din 89 de pacienţi cu gastrojejunostomie Y-Roux după rezecţie gastrică distală cu diverse indicaţii, am selecţionat 29 de pacienţi la care procedeul s-a folosit în tratamentul BARP. Am exclus 9 cazuri cu DDT pentru patologie primară de reflux alcalin. În 20 cazuri DDT a fost practicată ca o modalitate reconstructivă pe “stomac operat”: degastrogastrectomie sau conversie a montajului anastomotic preexistent (la bolnavi cu 1-3 operaţii în antecedente, cu tulburări severe de motilitate). Se constată o scădere a numărului de cazuri în ultimii ani. La 9 pacienţi DDT a fost utilizată ca intenţie curativă antireflux după chirurgia biliară:colecistectomie ± coledocoduodenostomie, constatând creşterea numărului de cazuri în ultima perioadă. Criteriile de indicaţie chirurgicală: clinice, radiologice, endoscopice, histologice au selecţionat pentru intervenţie cazurile severe. Se prezintă particularităţile tehnice ca şi consecinţele morfofuncţionale ale DDT. Rezultatele imediate sunt foarte bune: morbiditate minimă (o reintervenţie precoce pentru ocluzie digestivă înaltă) şi mortalitate postoperatorie zero. Rezultatele la distanţă –evaluate clinic, radiologic, endoscopic şi histologic- arată o ameliorare postoperatorie certă, cu excepţia anumitor forme histologice. Concluzii: Incidenţa BARP după chirurgia gastrică a scăzut, prin scăderea drastică a indicaţiei operatorii pentru boala ulceroasă; în schimb creşte relativ incidenţa acestei entităţi după chirurgia biliară. DDT este o procedură eficientă dar de rezervă, indicată în cazuri bine selecţionate. Se constată o ameliorare postoperatorie certa clinică, endoscopică şi histologică, cu excepţia gastritei atrofice şi a metaplaziei intestinale, care se ameliorează în mică măsură.Introduction. We analyzed the experience of the Clinic on past 30 years (1981-2010) regarding total duodenal diversion (TDD) with Roux- en- Y gastrectomy for postoperator alkaline reflux disease (PARD). Materials and method, results: Among 89 patients presenting Y-Roux gastrojejunostomy after gastric distal resection for various indications, we selected 29 patients in which the procedure was used as treatment of PARD. We excluded 9 patients with TDD for primary alkaline reflux disease. In 20 cases TDD was used as a reconstructive procedure on “operated stomach” : degastrogastrectomy or conversion of the existing anastomotic assembly (at patients with history of 1-3 gastric operations, with severe motility disorders). It is ascertained a decrease in the number of such cases in recent years. At another 9 patients TDD was used as an antireflux cure after biliary surgery: colecistectomy ± choledocoduodenostomy, noting the increase number of such cases lately. The criteria for surgery indication: clinicals, radiologycals, endoscopicals, histologicals selected for intervention severe cases. There are presentated techniques particularities and morfofunctional consequences of TDD. Immediate results were very good: minimal morbidity ( one early reintervention for acute digestive occlusion) and no postoperator mortality. Long time results – clinical, radiological, endoscopic and histological evaluated- showed a certain postoperator improvement, excepting some definite histological forms. Conclusions: PARD incidence after gastric surgery has decreased through drastically decrease of surgical indication for patients with gastroduodenal ulcer; after biliar surgery. TDD is an efficient procedure but as a backup, being indicated only in cases very carefully selected. It is observed a definite clinical, endoscopic and histological postoperator improvement excepting atrophic gastritis which is less improved

Advances and innovations in thoracic esophageal cancer surgery

Cancerul esofagian este o afecțiune malignă agresivă cu o incidență în continuă creștere și un prognostic nefavorabil. Tratamentul cancerului esofagian a devenit mai eficient in prezent prin abordarea multidisciplinară și prin crearea unor centre de excelență cu un volum mare de patologie esofagiană. Progresele în stadializare, tehnologia chirurgicală, terapia neoadjuvantă și îngrijirea perioperatorie au determinat reducerea morbidității și a mortalității. Principiul de bază actual al tratamentului curativ pentru boala localizată este intervenția chirurgicală, asociată cu radi-ochimioterapia neoadjuvantă pentru stadiile avansate local. Pentru a reduce morbiditatea postoper-atorie, au fost introduse în urmă cu 32 de ani, în arsenalul terapeutic al cancerului esofagian și tehnicile chirurgicale minim invazive. Există însă controverse legate de utilizarea abordului min-im invaziv în practică deoarece necesită o bază tehnică pretențioasă și dificil de accesat, o tehnică operatorie laborioasă și are o curbă lungă de învățare. Utilizarea tehnicilor chirurgicale minim invazive în tratamentul cancerului esofagian toracic, a avut totuși un impact important asupra morbidității și mortalității post-esofagectomie.Esophageal cancer is an aggressive malignancy with an increasing incidence and an unfavorable prognosis. The treatment of esophageal cancer has become more effective nowadays through the multidisciplinary approach and the creation of centers of excellence with a large volume of esoph-ageal pathology. Advances in staging, surgical technology, neoadjuvant therapy, and perioperative care have reduced morbidity and mortality. The current basic principle of curative treatment for localized disease is surgery, associated with neoadjuvant chemoradiotherapy for locally advanced stages. To reduce postoperative morbidity, minimally invasive surgical techniques and surgical techniques were introduced 32 years ago in the therapeutic arsenal of esophageal cancer. However, there is controversy about the use of the minimally invasive approach in practice because it re-quires a demanding and difficult to access technical basis, a laborious surgical technique and a long learning curve. The use of minimally invasive surgical techniques in the treatment of thoracic esophageal cancer, however, had a significant impact on post-esophagectomy morbidity and mortality

Prevalence of canine heartworm infection in Queensland, Australia: comparison of diagnostic methods and investigation of factors associated with reduction in antigen detection

Background: The prevalence of Dirofilaria immitis infection in dogs is increasing globally and spreading into new areas. Prevalence of dirofilariosis in the state of Queensland, Australia, was as high as 90% before the introduction of macrocyclic lactones. Limited research on prevalence of D. immitis infection in dogs in Queensland has been reported in the last 30 years. Antigen testing is the most common method for detection of dirofilariosis but its accuracy is reduced by antigen getting trapped (blocked antigen) in immune complexes (ICs). The objectives of this research were to determine the prevalence of D. immitis infection in dogs from two geographical areas (Brisbane and Townsville) in Queensland, to determine the extent to which blocked antigen affects the validity of antigen testing, and to explore whether this was associated with microfilaraemia, location, age or sex. Methods: Blood samples from Brisbane (sub-tropical climate) and Townsville (tropical climate) shelter dogs were evaluated for the presence of D. immitis antigen before (conventional antigen testing, CAT) and after dissociation of ICs by heat treatment (antigen testing after heat treatment, ATHT), using a commercially available test. Microfilariae were detected using modified Knott’s test (MKT). Test proportions were compared with McNemar’s test and the association between antigen test-discordant results (positive for antigen after dissociation of ICs) and microfilaraemia, location, sex and age was modelled using logistic regression. Results: Dirofilaria immitis prevalence in dogs from Townsville (22% by CAT, 32.1% by ATHT and 16.7% by MKT) was significantly higher than in dogs from Brisbane (1.1% by CAT and MKT and 1.7% by ATHT) (P<0.001). Dissociation of ICs allowed detection of significantly more D. immitis infected dogs than either conventional antigen testing or microfilariae detection, or the combined antigen and microfilariae detection (P<0.001). The odds of dogs being positive for antigen after dissociation of ICs were significantly higher for microfilaraemic, 3–4-year-old female dogs from Townsville. Conclusions: The high prevalence of infection with D. immitis in dogs from Townsville poses a health risk for local susceptible host species, including humans. Dissociation of ICs increases antigen detection and should be considered in dogs suspected of D. immitis infection but negative on routine testing

Development and Validation of Novel PCR Assays for the Diagnosis of Bovine Stephanofilariasis and Detection of Stephanofilaria sp. Nematodes in Vector Flies

Background: Stephanofilaria spp. nematodes are associated with cutaneous lesions in cattle and other livestock and mammalian wildlife species. In Australia, Haematobia irritans exigua, commonly known as buffalo fly (BF) transmits a well-described but presently unnamed species of Stephanofilaria, which has been speculatively implicated in the aetiology of BF lesions. The sensitivity of current techniques for detecting Stephanofilaria spp. in skin lesions and vector species is low, and there is no genomic sequence for any member of the genus Stephanofilaria currently available in sequence databases. Methods: To develop molecular assays for the detection of the Australian Stephanofilaria sp., skin biopsies were collected from freshly slaughtered cattle with typical lesions near the medial canthus. Adult nematodes and microfilariae were isolated from the biopsies using a saline recovery technique. The nematodes were morphologically identified as Stephanofilaria sp. by scanning electron microscopy. DNA was extracted and the internal transcribed spacer 2 (ITS2) region of rDNA, and the cytochrome c oxidase subunit 1 (cox1) region of mtDNA was amplified and sequenced. Stephanofilaria sp. specific polymerase chain reaction (PCR) and qPCR assays (SYBR Green® and TaqMan™) were developed and optimised from the novel ITS2 sequence obtained. The specificity of each assay was confirmed by testing against nematode species Onchocerca gibsoni and Dirofilaria immitis, as well as host (bovine) and BF DNA. Results: Scanning electron microscopy of the anterior and posterior ends of isolated nematodes confirmed Stephanofilaria sp. A phylogenetic analysis of the cox1 sequence demonstrated that this species is most closely related to Thelazia callipaeda, a parasitic nematode that is a common cause of thelaziasis (or eyeworm infestation) in humans, dogs, and cats. Both conventional and qPCR assays specifically amplified DNA from Stephanofilaria sp. Conventional PCR, TaqMan™, and SYBR Green® assays were shown to detect 1 ng, 1 pg, and 100 fg of Stephanofilaria DNA, respectively. Both qPCR assays detected DNA from single Stephanofilaria microfilaria. Conclusion: Molecular diagnostic assays developed in this study showed high specificity and sensitivity for Stephanofilaria sp. DNA. The availability of an accurate and sensitive PCR assay for Stephanofilaria will assist in determining its role in the pathogenesis of cattle skin lesions, as well as in understanding its epidemiological dynamics. This assay may also have application for use in epidemiological studies with other species of Stephanofilaria, most particularly closely related S. stilesi, but this will require confirmation

Immuno-fluorescence staining patterns of leukocyte subsets in the skin of taurine and indicine cattle

The immuno-staining patterns of skin leukocytes were investigated in three breeds of cattle: Holstein-Friesian, Brahman and Santa Gertrudis of similar age before and after tick infestation. The antibodies specific for CD45 and CD45RO reacted with cells in the skin of all Holstein-Friesian cattle but did not react with cells in the skin of any Brahman cattle. The same antibodies reacted with cells from the skin of four (CD45) and seven (CD45RO) of twelve Santa Gertrudis cattle. The antibodies specific for T cells and γδ subset of T cells recognized cells from all three breeds of cattle. The antibody specific for MHC class II molecules labelled cells of mostly irregular shape, presumably dermal dendritic cells and/or macrophages and Langerhans cells. The antibody specific for granulocytes (mAb CH138) reacted with cells only in sections cut from skin with lesions. The antibody specific for CD25+ cells labelled regularly shaped cells that showed a wide range of intensities of staining

Role of Staphylococcus agnetis and Staphylococcus hyicus in the Pathogenesis of Buffalo Fly Skin Lesions in Cattle

Buffalo flies (Haematobia irritans exigua) are hematophagous ectoparasites of cattle causing production and welfare impacts in northern Australian herds. Skin lesions associated with buffalo fly infestation and Stephanofilaria nematode infection are manifested as focal dermatitis or ulcerated areas, most commonly on the medial canthus of the eye, along the lateral and ventral neck, and on the abdomen of cattle. For closely related horn flies (Haematobia irritans irritans), Staphylococcus aureus has been suggested as a contributing factor in the development of lesions. To investigate the potential role of bacterial infection in the pathogenesis of buffalo fly lesions, swabs were taken from lesions and normal skin, and bacteria were also isolated from surface washings of buffalo flies and surface-sterilized homogenized flies. Bacterial identification was conducted by matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) and strain typing by repetitive sequence-based PCR (rep-PCR) and DNA sequencing to determine species similarity and virulence factors. Of 50 bacterial isolates collected from lesions, 38 were identified as Staphylococcus agnetis and 12 as Staphylococcus hyicus, whereas four isolates from normal skin were S. hyicus and one was Mammaliicoccus sciuri. Of the Staphylococcus isolates isolated from buffalo flies, five were identified as S. agnetis and three as S. hyicus. Fifty percent of the buffalo fly isolates had rep-PCR genotypic patterns identical to those of the lesion isolates. Genome sequencing of 16 S. agnetis and four S. hyicus isolates revealed closely similar virulence factor profiles, with all isolates possessing exfoliative toxin A and C genes. The findings from this study suggest the involvement of S. agnetis and S. hyicus in buffalo fly lesion pathogenesis. This should be taken into account in the development of effective treatment and control strategies for lesions