Migratory Tracts Research Articles

Development of an intraventricular adeno-associated virus-based labeling strategy for glioblastoma cells nested in the subventricular zone.

Glioblastoma (GBM) is a dreadful brain tumor, with a particular relationship to the adult subventricular zone (SVZ) that has been described as relevant to disease initiation, progression, and recurrence. We propose a novel strategy for the detection and tracking of xenografted GBM cells that are located in the SVZ, based on an intracerebroventricular (icv) recombinant adeno-associated virus (AAV)-mediated color conversion method. We used different patient-derived GBM stem-like cells (GSCs), which we transduced first with a retroviral vector (LRLG) that included a lox-dsRed-STOP-lox cassette, upstream of the eGFP gene, then with rAAVs expressing the Cre-recombinase. Red and green fluorescence is analyzed in vitro and in vivo using flow cytometry and fluorescence microscopy. After comparing the efficiency of diverse rAAV serotypes, we confirmed that the in vitro transduction of GSC-LRLG with rAAV-Cre induced a switch from red to green fluorescence. In parallel, we verified that rAAV transduction was confined to the walls of the lateral ventricles. We, therefore, applied this conversion approach in 2 patient-derived orthotopic GSC xenograft models and showed that the icv injection of an rAAV-DJ-Cre after GSC-LRLG tumor implantation triggered the conversion of red GSCs to green, in the periventricular region. Green GSCs were also found at distant places, including the migratory tract and the tumor core. This study not only sheds light on the putative outcome of SVZ-nested GBM cells but also shows that icv injection of rAAV vectors allows to transduce and potentially modulate gene expression in hard-to-reach GBM cells of the periventricular area.

Indigenous technical knowledge used in dairying by pastoralists of Jammuand Kashmir

The study was carried out in current Union Territory of Jammu & Kashmir (erstwhile state of Jammu & Kashmir). Two districts, namely Rajouri & Poonch from Jammu division were selected purposively because of being the districts of highest Gujjar (transhumants primarily rearing cattle and buffaloes)population; whereas, two districts, namely Anantnag & Baramullafrom Kashmir division were selected purposively because of beingthe districts of highest Bakarwal (transhumants primarilyrearing sheep and goats) population. Sixty respondents (Gujjars)from migratory tracts of district Rajouri & Poonch from Jammudivision were selected randomly and sixty respondents(Bakarwal) from migratory tract of districts Anantnag &Baramulla from Kashmir division were selected randomly comprising a total of 120 respondents. Those pastoralists who were rearing at least 10 milch animals were selected for the study. Focus group discussion method was used for collection of data related to Indigenous Technical Knowledges (ITKs). The discussion revealed that the respondents were using ITKs primarily in breeding, feeding and health-care management practices. With regard to the induction of heat, thirty per cent of the respondents used chapatti of a mixture of Gur and ghee inthe ratio of 2:1 in wheat flour and fed to animal once a day toinduce heat. For retention of placenta, 34.17 per cent of therespondents fed extract obtained from tea leaves (Camelliasinensis) on boiling after sieving to animals. As far as ITKsregarding feeding practices are concerned 40.83 per cent of therespondents fed boiled Ber leaves (Ziziphus mauritiana) onceaday to increase milk production. The study reveals that 42.50 percent of respondents fed10-12 leaves of Katori(Xylosonalongifolium) plant to animals having diarrhoea and15.83 per cent fed fruits of bel (Aegle marmelos) to animals having diarrhoea.

CT can identify characteristic features of hypaxial muscle abscesses in dogs due to presumed migrating vegetal foreign material as well as additional changes along the migratory tract in other anatomic regions.

Hypaxial muscle abscess is an important differential in dogs presenting for abdominal or back pain, lameness, and nonspecific signs like fever, lethargy, and hyporexia. It can occur concurrently with intrathoracic disease such as pyothorax secondary to migrating vegetal foreign material. Twelve dogs that underwent CT of the lumbar spine or abdomen and had a diagnosed hypaxial abscess on surgical and/or microbiological examination were included in this retrospective, descriptive case series. Computed tomography findings and findings from other imaging modalities employed were described. Eleven dogs were hunting breeds. Clinical signs included lethargy, fever, increased respiratory effort, and abdominal or back pain. Radiography and/or ultrasonography were employed during preliminary work up at clinician discretion and respectively revealed changes consistent with osteomyelitis in the cranial lumbar vertebrae and heterogenous, hypoechoic areas in the hypaxial musculature consistent with abscesses. Computed tomography findings included enlargement of hypaxial muscles with well-defined fluid attenuating noncontrast enhancing areas with a contrast-enhancing rim consistent with abscesses, periosteal reaction and lysis of vertebrae, and retroperitoneal effusion. Four of the 12 cases in this series had material identified and removed at surgery. The other eight cases were presumed to be the same disease process based on compatible signalment, imaging findings, and microbiological results. Migrating vegetal foreign bodies are a common problem at the authors' institution. Computed tomography provided expedient, thorough visualization of the relevant hypaxial lesions for diagnostic and surgical planning purposes and also characterized intrathoracic components of this disease.

Sheep pastoralism in southern Karnataka

Present study was conducted to understand and document the migratory system followed, socio-economic status, management and healthcare practices adopted by pastoralists of southern Karnataka. In-person interview method with a structured questionnaire was used for collecting data from 25 shepherd groups on migration. Migration starts during December and continues for 6–8 months covering around 200 km. The breed is locally called Tumkur or Sira sheep which is not yet recognized as a separate breed. Migratory flock usually contains 1,000 to 1,200 sheep, 50 goats, 10 donkeys and 2–4 dogs. The average adult male to female ratio is 1 : 22. Sale of lambs and penning rental are the main sources of income. Sheep pastoralism is an established and effective way of utilizing the excess green cover in transient regions by the sheep from pasture deficient dry zones. Maintaining the social and community pasture and forest lands at their home place will be a benefit during the lean and non-migratory periods. Assessing the migratory paths, fodder and water resource and penning demand by the farmers on migratory tract will help policy makers to provide suitable interventions to curtail the problems of pastoralists. The traditional knowledge of shepherds should be documentedand validated for larger use. The pastoralists need to be educated and trained about the use of digital finance management, herd diversification, health management and government schemes to increase their profitability and standard of living.

Anatomic pathology of cattle and goat liver collected from slaughterhouses in Mymensingh city of Bangladesh

The morphologic abnormality of cattle and goat livers (collected from slaughterhouses in Mymensinghcity of Bangladesh) were investigated in the present study. The livers of slaughtered cattle (n = 14) and goat (n = 20) were examined by gross visualization, palpation and incision as well as histopathology. The total liver lesion in cattle was 71.4% and in goat was 75%. Grossly, various pathological changes, mainly thickened, rough capsule which was whitish in color, necrotic foci, thickened bile ducts with matured flukes, fibrosis were seen. Histopathologically, necrosis, degenerative changes of hepatocytes, dilated sinusoids, inflammatory infiltrates, granuloma, bile duct hyperplasia, periportal and periductal fibrosis were found in cattle and goat livers. Extensive fibrosis was seen portal areas of liver by Goldner’s trichrome staining. Migratory tracts and immature flukes were seen in intrahepatic bile ducts. Most common cause of liver lesions found in the present study was Fascioliasis in both cattle and goat. It could be concluded that the major gross and microscopic alterationsin the livers of cattle and goat due to Fasciola giganticainfectionreflected tissue damage, which can lead to substantial financial losses in animals and great health problems in human. Therefore, special care and attention are required to ensure that seriously damaged livers of food animals are not passed on for human consumption, and extensive monitoring and epidemiological surveys are necessary to cope this adverse situation

Acute fasciolosis outbreak with aberrant larval migration among sheep in Kashmir Valley

The present study describes the outbreak of acute fasciolosis among sheep in the twin districts of Budgam and Kupwara of the Kashmir Valley in 2019-20. To ascertain the cause of deaths, clinical examination and fecal examination of diseased sheep and post-mortem examination of the dead animals was carried out. The diseased sheep presented symptoms of cold extremities, anorexia, dullness, aloofness, depression, green-colored profuse watery discharge from nose and mouth (rumen contents) and recumbency before death. The liver showed typical migratory tracts due to fasciolosis. Fasciola spp. parasites were observed in liver and gall bladder during postmortem examination. The aberrant migration of the parasite was also noted in some cases with adult parasites recovered from lungs in only one case. All fecal samples tested negative for Fasciola spp. eggs. Most of the deaths were observed in pregnant ewes and younger sheep. Further mortality was controlled by mass dosing with Triclabendazole. The results indicated Acute Fasciolosis as the cause of death in these sheep as revealed by gross lesions and retrieval of adult Fasciola spp. from liver and gallbladder. The parasite demands serious attention due to its ill effects on animal health, economy of farmers and zoonotic implications.

Molecular characterization and histopathological studies on Fasciola gigantica in Mithun (Bos frontalis)

Mitochondrial DNA sequence of the sub-unit 1 of cytochrome c oxidase (Cox1) and ribosomal DNA sequence of the first internal transcribed spacer (ITS-1) of Fasciola collected from mithun and cattle from Arunachal Pradesh, India were characterized at molecular level in order to identify the species of parasite. Based on sequence and phylogenetic analysis, the identity of the parasite was confirmed as Fasciola gigantica in mithun as well as from cattle. In order to know histopathological alteration in Fasciola infection, a histopathological study was performed on eight liver specimens, out of 110 animals studied during the period from 2010-2016. Histopathological examination of F.gigantica infected liver showed extensive fibrous connective tissue proliferation with necrosis of hepatocytes and infiltration of polymorphonuclear cell. There was evidence of migratory tracts of parasites with losing of normal lobular hepatic architecture. Bile duct proliferation was followed by congestion of portal vein with perivascular cuffing with surrounding degeneration of hepatocytes. The hepatocytes showed pyknosis with hyperplasia of bile duct.

Occurrence of Fasciola spp. in free-ranging wild ruminants of Central India

Wild ruminants are prime source for survival of apex species in protected and nonprotected forest areas. During post mortem examination of deer, antelopes and gaurs in different tiger reserves of central India, the occurrence of Fasciola spp. was documented. Out of 155 carcasses, 48 (30.9%) harboured liver flukes including chital (42.1%) blackbuck (35.2%), nilgai (28.5%), chinkara (25.0%), gaur (25%), sambar (18.7%) and barking deer (12.5%). In some cases, extensive destruction in liver parenchyma was recorded with migratory tracts, nodules and necrosis. The recovered flukes (41.7±2.26 x 5.12±017 mm) and eggs (171± 3.46 x 89.2± 1.48 μm) were measured and confirmed as Fasciola gigantica. The possible reasons for the varying degrees of infection with Fasciola in different wild ruminants is discussed.

Can Medullary Thyroid Carcinoma Arise in Thyroglossal Duct Cysts? A Search for Parafollicular C-cells in 41 Resected Cases.

Thyroglossal duct cysts (TGDCs) are present in ~7% of adults and develop from the midline migratory tract between the foramen cecum and anatomic location of the thyroid. Thyroid tissue can be identified in 2/3 of TGDCs, and up to 1% develop associated malignancy, 90% of which are papillary thyroid carcinoma. Cases of follicular and anaplastic carcinoma have been documented, but there are no reports of medullary thyroid carcinoma arising in a TGDC. This is presumably due to the distinct embryologic origin of parafollicular C-cells, from which medullary carcinoma arises. The goal of this study is to determine whether parafollicular C-cells are present in TGDCs. H&E sections from 41 TGDC cases were examined for thyroid tissue, thyroglossal duct remnants, ultimobranchial remnants, and parafollicular C-cells. Immunohistochemistry was performed for TTF-1 and calcitonin. Eighty three percent (34/41) of cases contained thyroid tissue on H&E and by TTF-1. No cases (0/41) had ultimobranchial remnants or parafollicular C-cells on H&E or with calcitonin. One case of papillary carcinoma in a TGDC was identified. These cases illustrate that although TGDCs often contain thyroid tissue, parafollicular C-cells are absent. Therefore, unlike other thyroid neoplasms, there is no evidence to support the possibility of medullary carcinoma arising in a TGDC.

Patho-Surveillance and Pathology of Fascioliosis (Fasciola gigantica) in Black Bengal Goats in Bangladesh

The aim of the study was conducted to investigate patho-surveillance and pathology of fascioliosis in Black Bengal goats in Sylhet region of Bangladesh. Patho-surveillance and Pathology on fascioliasis of Black Bengal goats were undertaken in such localities under different climatic conditions existing in Sylhet region of five different Upazilla. Goats were divided into young and adult groups. Affected liver samples from slaughtered goats were collected randomly and examined by grossly and histopathological study for hematoxylin and eosin staining. A total of 2000 Black Bengal goat livers were examined, of which 202(10.10%) livers were found to be affected with fascioliosis. Prevalence of fascioliosis was significantly higher in young goats (15.58 %) than in adult (9.59%) and female goats (13.10%) were more susceptible than male goats (7.10%). Seasonal prevalence of fascioliosis was also studied. Highest prevalence (16.51 %) was recorded during rainy season and lowest prevalence (4.70%) was recorded during summer season. Grossly, affected livers were enlarged with rounded edges and thickened capsule. In acute cases, numerous haemorrhagic spots were found on the parietal & visceral surfaces of the affected liver. In chronic form, liver was cirrhotic and reduced in size. The affected intra-hepatic bile ducts were protruded and engorged with flukes. Microscopically, migratory tracts were represented by the presence of haemorrhage, edema and infiltration of numerous eosinophils mixed with few lymphocytes. Fatty change, atrophy and necrosis of hepatocytes were recorded along with deposition of bile pigment in hepatic parenchyma and damage of portal tract area. The wall of bile ducts was thickened due to fibrosis and lining epithelia were hyperplastic. Cross sections of adult and immature flukes were found within the lumen of the thickened bile ducts and hepatic parenchyma respectively. The study to provides valuable in insight towards better understanding of epidemiology and pathogenesis of the disease in Black Bengal goats to adopt suitable control strategies against the disease and meat inspection at slaughter houses.

An outbreak of acute fasciolosis in goats of Chhattisgarh

An outbreak of acute fasciolosis occurred at Goat Breeding Farm, Pakaria, Bilaspur, Chhattisgarh. Out of 416 goats, 36 goats (21 females and 15 males) exhibiting symptoms of dullness, weakness, anorexia and paleness of visible mucous membrane died within a time span of 16 days. The mortality rate in male goats was 16.55% compared to 6.50% in females. The post-mortem examination of dead animals revealed extensive haemorrhage and marked destruction of liver parenchyma. The mean haematological values of RBC, Hb and PCV were decreased in clinically infected goats. The mean values of total serum protein and albumin were also decreased in infected animals. In addition, the mean biochemical values of serum aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase in sick animals were higher. The histopathological changes in the livers of infected animals revealed excessive proliferation of fibrous tissue around the bile duct along with infiltration of numerous eosinophils admixed with lymphocytes. Sections of fluke remnants surrounded by clear spaces were found in the dilated and thickened bile ducts. The migratory tracts of the immature liver flukes were filled with necrosed hepatocytes and there was infiltration of eosinophils with lymphocytes along with haemorrhage and proliferation of fibrous tissue. The migrating flukes produced severe reaction in the host tissue and in the later stages they became surrounded by macrophages, eosinophils and granular tissue.

Morphological, molecular and pathological appraisal of Callitetrarhynchus gracilis plerocerci (Lacistorhynchidae) infecting Atlantic little tunny (Euthynnus alletteratus) in Southeastern Mediterranean

The Atlantic little tunny, Euthynnus alletteratus, is widely distributed in temperate and tropical waters of the Atlantic Ocean, Black and Mediterranean Seas. In this study, wild-caught little tunny from Egypt, were found to be naturally infected with trypanorhyncha metacestodes, and the overall prevalence rate of infection was 38.7%. The blastocysts were either loosely attached to the mesentery of infected fish, or firmly attached and deeply embedded within the hepatic parenchyma. These encysted plerocerci are identified as Callitetrarhynchus gracilis (Trypanorhyncha, Lacistorhynchidae) based on its morphological and molecular characterization. The morphological characteristics of C. gracilis including scolex shape; the bothridia groove; the presence of frontal glands; the length of post-larval (appendix); metabasal armature; the existence of ‘Chainette’ and satellite hooks of different size were studied and described by Light and Scanning electron microscope. The phylogenetic analysis of lsrDNA gene of plerocerci confirmed the identification of the species to be deeply embedded in genus Callitetrarhynchus. The histopathological examination revealed severe pathological changes in the affected organs, including necrosis, inflammatory reactions, fibrosis and migratory tracts of the parasitic larvae together with marked visceral organs adhesions. To the best of our knowledge, this is the first report describing the detection of C. gracilis in little tunny collected from the Abu Qir landing site in Alexandria, Egypt.

Growth rate and survivability patterns in Jamunapari breeds of goats under farm conditions in Mahoba district of Bundelkhand region

The present investigation was carried out in the breeding and migratory tracts of jamunapari goat in district Mohoba of Bundelkhand region of Uttar Pradesh. For study, data were collected from 10 villages of belonging to Jaitpur block of the Mohoba district in 2012-2013. The climate of the breeding and migratory tracts was hot, semi-arid and tropical in nature with matching vegetation and cultivated crops. The average migratory flock size was 58 ± 3.71 (range 30-100). In stationary flocks the size ranged between 2 and 35 with an average of 17. The average flock contains 16.0 ± 2.57 individuals (range 8 to 41), of which 0.25% adult males, 8.65% adult females and 7.1% young. It has been observed that Feeding system significantly affected body weights and weight gain at all the growth stages and. In stationery tracts approximately diarrhea 21.7%, pneumonia 7.1%, parasitic disease 5.5%, F.M.D. 4.9%, Nutritional disease 10.6%, Blot 6.7%, P.P.R. 11.5%, enterotoxaemia 8.4% and unknown other diseases 5.9% were found. In Migratory tracts approximately diarrhea 24.5%, Phenomena 9.6%, Parasitic disease 7.8%, F.M.D. 6.6%, Nutritional disease 12.8%, Blot 8.2%, P.P.R. 13.7%, enterotoxaemia 10.1% and unknown other diseases 7.4%, which were found to have more than stationery tracts. The study revealed that jamunapari goat was reared under traditional method of rearing with less scientific management practices.

Pathological study of acute fasciolosis in goats in Karnataka

The present report pertains to the pathological investigation of anoutbreak of acute fasciolosis in goats in a private goat farm at Mysore, Karnataka. The farm comprised of 125 indigenous goats of all age groups reared under extensive system. Goats started dying without showing any clinical symptoms, except brief period of anorexia. Detailed necropsy was performed on 6 animals died over a span of 15 days. Thecarcasses were severely dehydrated, thin body condition with pallor oral and conjunctival mucosa. Post-mortem examination revealed about 500 ml of serosanguinous fluid and massive blood clots of varying sizesweighing up to of 500-600g in the abdominal cavity of few goats. The fluke migratory tracks were seen with severe haemorrhages on the liver surface. The cut surface of liver parenchyma and bile ducts showed immature liver flukes. The gall bladder was severely distended with blood, and large blood clotsin all the carcasses. The duodenal and large intestinal mucosa was moderately thickened and severely congested and clotted blood in the lumen of entire small intestine. The histological examination of liver exhibited extensive fibrosis, necrosisand haemorrhages with severe infiltration of mononuclear inflammatory cells, especially surrounding the parasites. There was mild proliferation of bile ducts around dead tissue with migratory tracts of hemosiderin. Pathological investigation confirmed the disease in goats as acute fasciolosis.

The remarkable migration of the medial collateral ligament

The developing cortical surfaces of long bones are sculpted and modeled by periosteal osteoclasts and osteoblasts. These surfaces also receive the insertions of tendons and ligaments, and these insertion sites too are modeled to form the root systems that anchor them into the cortical bone. The regulatory molecules that control modeling are poorly understood, but recent evidence suggests that parathyroid hormone-related protein (PTHrP) participates in this process. PTHrP functions principally as a paracrine regulatory molecule, and is known to be induced by mechanical loading in a number of sites. The most curious example of developmental modeling of the cortex is the migration of insertion sites such as that of the medial collateral ligament (MCL) along the bone surface during long-bone growth. We report here the mechanisms that mediate MCL migration using a combination of genetic, imaging and histological techniques. We describe a MCL migratory complex that comprises two components. The first is the MCL insertion site itself, which is a prototypical fibrous insertion site with coupled osteoclast and osteoblast activities, and its key feature is that it is anchored early in development, well before initiation of the long-bone growth spurt. Above the insertion site the periosteum is excavated by osteoclasts to form a migratory tract; this is mediated by wholly uncoupled osteoclastic bone resorption and remains as an unmineralized canal on the cortical surface in the adult. Load-induction of PTHrP appears to regulate the osteoclastic activity in both the insertion site and migratory tract.

Visceral larva migrans presenting as multiple liver abscesses

Eosinophilic necrotizing granulomas in the liver, known as visceral larva migrans (VLM), are a rare cause of liver abscesses. The term VLM refers to infections in human tissues caused by the larval stages of ascarid worms of dogs and cats. We report two cases of VLM which presented with high grade fever and abdominal pain for 3-4 months. Marked peripheral eosinophilia, multiple confluent perivenous lesions in the right lobe of liver with characteristic migratory tracts on imaging and biopsy evidence of necrotizing eosinophilic granulomas led to the diagnosis. Complete recovery was achieved with repeated courses of a combination of anthelmintics.

The pathology observed in experimental <i>Fasciola gigantica</i> infected Yankasa sheep in Zaria Nigeria.

A study was conducted to determine the pathogenic effects of Fasciola gigantica infection on Yankasa sheep for a period of four months (April-July ,2010). Pathological lesions were observed in four Fasciola gigantica infected Yankasa sheep that died at the 10th, 11th and 12th week post-infection in an experimental infection at the Reproduction unit of the National Animal production Research institute, Shika-Zaria, Nigeria. The experiment involved twelve Yankasa sheep that were divided into two groups of infected and controls. The six animals in the infected group were each orally inoculated with 1200 Fasciola gigantica metacercariae and monitored for a period of 16 weeks. The pathogenic effects of the Fasciola gigantica infection began to manifest through death of four sheep among the infected group; with one death observed on the 10th week, two on the 11th and one on the 12th week post-infections respectively. The gross pathological lesions observed were hepatomegally, appearance of migratory tracts on the liver surface, appearance of puncture wounds through which protruded the anterior ends of the flukes to the liver surface as well as a fluid filled cavity with each having a fluid content of not less than 2.5 litres within the abdominal cavity. Other features observed grossly were marked distension of the gall bladder in which numerous flukes were present. The histopathological lesions were presented inform of intense hemorrhage both in the parenchyma and in the parasite tracts. There was fibrosis and distortion of the normal architecture of the hepatic cells. Observed clinical signs were inappetance, progressive anaemia and emaciation. There was a marked reduction in albumin and total plasma protein levels in the blood of the infected sheep compared to their controls. The findings of this study revealed that Fasciola gigantica is highly pathogenic to Yankasa sheep, therefore strategic control of the parasite and its intermediate host in the study area is recommended for improved sheep production.Keywords: Yankasa sheep, Liver, lesion

INCIDENCE AND PATHOLOGICAL CHANGES IN FASCIOLIASIS (<i>FASCIOLA GIGANTICA</i>) OF DOMESTICATED DEER

An investigation was carried out on incidence and pathology of fascioliasis in deer of Chittagong Zoo and some houses of Chittagong city during the period from January 2001 to June 2004. A total of 57 domesticated deer of various types were examined to diagnose the fascioliasis in Chittagong zoo and some house deer of Chittagong city. By faecal examination out of 44 cases 34 (77.3%) were positive for fascioliasis, of which, 15 (34.1%) cases were recorded in Chittra deer, 12 (27.3%) in Maya deer, 05 (11.4%) in Shambar deer and 02 (4.5%) in Nathrini deer. In post mortem examination, fascioliasis found in 13 (100%) cases, of which, 08 (61.5%) from Chittra deer and 05 (38.5%) from Maya deer. The overall incidence of fascioliasis in deer was 82.5%. In relation to sex the fasciolasis was significantly (p < 0.05) higher in female (82.6%) than male (71.4%) deer. The incidence of fascioliasis in various types of deer in relation to age was significantly (p < 0.05) higher in age group of above 2 years old than in age group below 2 years old. The gross examination revealed enlarged livers with round edges and thickened capsule with numerous haemorrhagic spots on the parietal surface (subacute form). In chronic form, the livers were cirrhotic and reduced in size. The affected intra-hepatic bile ducts were protruded and were engorged with flukes. Microscopically the migratory tracts were represented by the presence of haemorrhagic, oedema and infiltration with numerous eosinophils mixed with few lymphocytes. The wall of the bile ducts was thickened with fibrous tissue proliferation and the lining epithelium showed hyperplastic changes.

Immunohistochemical characterization of lymphocyte and myeloid cell infiltrates in spirocercosis‐induced oesophageal nodules

Spirocerca lupi is a nematode that infects the dog's oesophagus and promotes the formation of an inflammatory fibroblastic nodule that progresses to sarcoma in approximately 25% of cases. Spirocercosis-associated oesophageal sarcoma is an excellent and under-utilized spontaneous model of parasite-associated malignancy. The inflammatory infiltrate of paraffin-embedded, non-neoplastic oesophageal nodules (n = 46), neoplastic nodules (n = 25) and normal oesophagus (n = 14) was examined by immunohistochemistry using MAC387 (myeloid cells), CD3 (T cells), Pax5 (B cells) and FoxP3 (T regulatory cells) antibodies. Myeloid cells predominated in 70% of nodules, in pockets around the worms' migratory tracts and in necro-ulcerative areas in neoplastic cases. T cells predominated in 23% of cases with a focal or diffuse distribution, in the nodule periphery. No significant differences were observed between neoplastic and non-neoplastic stages. FoxP3+ cells were observed in low numbers, not significantly different from the controls. The inflammation in spirocercosis is characterized by pockets of pus surrounded by organized lymphoid foci. There was no evidence of a local accumulation of FoxP3+ cells, unlike many previous studies that have reported an increase in FoxP3+ T cells in both malignancies and parasite infections. The triggering factor(s) driving the malignant transformation of the spirocercosis-associated chronic inflammatory nodule warrants further investigation.

Non-cysteine linked MUC1 cytoplasmic dimers are required for Src recruitment and ICAM-1 binding induced cell invasion

BackgroundThe mucin MUC1, a type I transmembrane glycoprotein, is overexpressed in breast cancer and has been correlated with increased metastasis. We were the first to report binding between MUC1 and Intercellular adhesion molecule-1 (ICAM-1), which is expressed on stromal and endothelial cells throughout the migratory tract of a metastasizing breast cancer cell. Subsequently, we found that MUC1/ICAM-1 binding results in pro-migratory calcium oscillations, cytoskeletal reorganization, and simulated transendothelial migration. These events were found to involve Src kinase, a non-receptor tyrosine kinase also implicated in breast cancer initiation and progression. Here, we further investigated the mechanism of MUC1/ICAM-1 signalling, focusing on the role of MUC1 dimerization in Src recruitment and pro-metastatic signalling.MethodsTo assay MUC1 dimerization, we used a chemical crosslinker which allowed for the detection of dimers on SDS-PAGE. We then generated MUC1 constructs containing an engineered domain which allowed for manipulation of dimerization status through the addition of ligands to the engineered domain. Following manipulation of dimerization, we immunoprecipitated MUC1 to investigate recruitment of Src, or assayed for our previously observed ICAM-1 binding induced events. To investigate the nature of MUC1 dimers, we used both non-reducing SDS-PAGE and generated a mutant construct lacking cysteine residues.ResultsWe first demonstrate that the previously observed MUC1/ICAM-1signalling events are dependent on the activity of Src kinase. We then report that MUC1 forms constitutive cytoplasmic domain dimers which are necessary for Src recruitment, ICAM-1 induced calcium oscillations and simulated transendothelial migration. The dimers are not covalently linked constitutively or following ICAM-1 binding. In contrast to previously published reports, we found that membrane proximal cysteine residues were not involved in dimerization or ICAM-1 induced signalling.ConclusionsOur data implicates non-cysteine linked MUC1 dimerization in cell signalling pathways required for cancer cell migration.