Three-dimensional culture model to study the biology of vacuolated notochordal cells from mouse nucleus pulposus explants.

Intervertebral disc degeneration (IDD) involves cellular changes in the nucleus pulposus (NP) characterised by a decline of the large vacuolated notochordal cells (vNCs) and a rise of smaller vacuole-free mature chondrocyte-like NP cells. An increasing number of studies demonstrate that notochordal cells (NCs) exert disease-modifying effects, establishing that NC-secreted factors are essential for the maintenance of a healthy intervertebral disc (IVD). However, understanding the role of the NCs is hampered by a restricted reserve of native cells and the lack of robust ex vivo cell model. A precise dissection enabled the isolation of NP cells from 4 d post-natal stage mouse spines and their culture into self-organised micromasses. The maintenance of cells' phenotypic characteristics was demonstrated by the presence of intracytoplasmic vacuoles and the immuno-colocalisation of the NC-markers (brachyury; SOX9) after 9 d of culture both in hypoxic and normoxic conditions. A significant increase of the size of the micromass was observed under hypoxia, consistent with a higher level of Ki-67+ immunostained proliferative cells. Furthermore, several proteins of interest for the study of vNCs phenotype (CD44; caveolin-1; aquaporin 2; patched-1) were successfully detected at the plasma membrane of NP-cells cultured in micromasses under hypoxic condition. IHC was performed on mouse IVD sections as control staining. An innovative 3D culture model of vNCs derived from mouse postnatal NP is proposed, allowing future ex vivo exploration of their basic biology and of the signalling pathways involved in IVD homeostasis that may be relevant for disc repair.

Tetra disseminated microsporidiosis: a novel disease in ornamental fish caused by Fusasporis stethaprioni n. gen. n. sp.

A novel microsporidial disease was documented in two ornamental fish species, black tetra Gymnocorymbus ternetzi Boulenger 1895 and cardinal tetra Paracheirodon axelrodi Schultz 1956. The non-xenoma-forming microsporidium occurred diffusely in most internal organs and the gill, thus referring to the condition as tetra disseminated microsporidiosis (TDM). The occurrence of TDM in black tetra was associated with chronic mortality in a domestic farmed population, while the case in cardinal tetra occurred in moribund fish while in quarantine at a public aquarium. Histology showed that coelomic visceral organs were frequently necrotic and severely disrupted by extensive infiltrates of macrophages. Infected macrophages were presumed responsible for the dissemination of spores throughout the body. Ultrastructural characteristics of the parasite developmental cycle included uninucleate meronts directly in the host cell cytoplasm. Sporonts were bi-nucleated as a result of karyokinesis and a parasite-produced sporophorous vesicle (SPV) became apparent at this stage. Cytokinesis resulted in two spores forming within each SPV. Spores were uniform in size, measuring about 3.9 ± 0.33 long by 2.0 ± 0.2 µm wide. Ultrastructure demonstrated two spore types, one with 9-12 polar filament coils and a double-layered exospore and a second type with 4-7 polar filament coils and a homogenously electron-dense exospore, with differences perhaps related to parasite transmission mechanisms. The 16S rDNA sequences showed closest identity to the genus Glugea (≈ 92%), though the developmental cycle, specifically being a non-xenoma-forming species and having two spores forming within a SPV, did not fit within the genus. Based on combined phylogenetic and ultrastructural characteristics, a new genus (Fusasporis) is proposed, with F. stethaprioni n. gen. n. sp. as the type species.

[An unusual cause of a decline in general health in a very old person]. / Une cause inhabituelle d'altération de l'état général chez une personne très âgée.

Humans are accidental intermediate hosts and a dead-end for the echinococcosis parasite. No cases of hydatid echinococcosis (hydatid cyst) have ever been described in a French-born very elderly person, who has never been out of France. We report the case of a 90-year-old man hospitalised following a decline in his general health and the discovery of a liver mass on the CT-scan. The abdominal palpation was painless, and no mass was felt. Biological tests revealed hypereosinophilia, a mild inflammatory syndrome, a slight increase in the immunoglobulin E level, and positive serology for Echinococcus granulosus. A diagnosis of hydatid echinococcosis due to Echinococcus granulosus was made given the liver mass on the CT-scan, the positive serology and the hypereosinophilia. Management consisted of watch and wait. This observation is interesting because it was a case of an autochthonous hydatid cyst of an unexpected discovery in a very elderly patient. Indeed, even though the discovery was classical, the clinical presentation was singular because of the context.

L'être humain est un hôte intermédiaire accidentel et une impasse parasitaire pour l'échinococcose. Aucun cas de kyste hydatique (échinococcose hydatique) n'a été décrit chez une personne très âgée née en France et ne l'ayant jamais quittée. Nous rapportons ici le cas d'un patient âgé de 90 ans hospitalisé pour altération de l'état général et découverte d'une masse hépatique à la tomodensitométrie. La palpation abdominale était indolore sans masse perçue. Le bilan biologique révélait une hyperéosinophilie, un discret syndrome inflammatoire, une légère augmentation du taux d'immunoglobuline E et une sérologie de l'Echinococcus granulosus positive. Le diagnostic d'échinococcose hydatique (kyste hydatique) due à Echinococcus granulosus fut alors posé, compte tenu de la masse hépatique découverte à la tomodensitométrie, de la sérologie positive et de l'hyperéosinophilie. La prise en charge a consisté en une attitude «watch and wait¼. Cette observation est intéressante car il s'agissait d'un cas de kyste hydatique autochtone de découverte fortuite chez un patient très âgé. En effet, même si le mode de découverte était classique, le tableau clinique était singulier en raison du contexte.

[Anaplastic large cell lymphoma associated with breast implants, diagnosed by fine needle aspiration. Report of one case]. / Linfoma anaplásico de células grandes asociado a implantes mamarios diagnosticado mediante punción por aguja fina. Caso clínico.

Breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) is a rare type of non-Hodgkin T-cell lymphoma, recently defined in the 2016 World Health Organization (WHO) classification of lymphoid neoplasms. It occurs more commonly when textured implants are used and appears clinically as a late seroma. Cytologically, these lesions are composed of large atypical cells with pleomorphic nucleus and an immunophenotype positive for T cell markers and CD30, and negative for ALK1. We report a 56-years-old woman with breast implants who developed a periprosthetic seroma three years after surgery. A fine needle aspiration of the lesion was carried out. Cytology and the immunocytochemical study revealed cells compatible with BIA-ALCL. The flow cytometric study was negative. Excisional biopsy of the capsule was performed, observing that the neoplastic cells were confined to the inner surface of the capsule. Imaging studies did not find evidence of disseminated disease. The present case demonstrates the importance of the study of any late periprosthetic effusion, which can be performed using fine needle aspiration.

Characterization of spaC-type Erysipelothrix sp. isolates causing systemic disease in ornamental fish.

Since 2012, low-to-moderate mortality associated with an Erysipelothrix sp. bacterium has been reported in ornamental fish. Histological findings have included facial cellulitis, necrotizing dermatitis and myositis, and disseminated coelomitis with abundant intralesional Gram-positive bacterial colonies. Sixteen Erysipelothrix sp. isolates identified phenotypically as E. rhusiopathiae were recovered from diseased cyprinid and characid fish. Similar clinical and histological changes were also observed in zebrafish, Danio rerio, challenged by intracoelomic injection. The Erysipelothrix sp. isolates from ornamental fish were compared phenotypically and genetically to E. rhusiopathiae and E. tonsillarum isolates recovered from aquatic and terrestrial animals from multiple facilities. Results demonstrated that isolates from diseased fish were largely clonal and divergent from E. rhusiopathiae and E. tonsillarum isolates from normal fish skin, marine mammals and terrestrial animals. All ornamental fish isolates were PCR positive for spaC, with marked genetic divergence (<92% similarity at gyrB, <60% similarity by rep-PCR) between the ornamental fish isolates and other Erysipelothrix spp. isolates. This study supports previous work citing the genetic variability of Erysipelothrix spp. spa types and suggests isolates from diseased ornamental fish may represent a genetically distinct species.

Pathologic features of mycobacteriosis in naturally infected Syngnathidae and novel transcriptome assembly in association with disease.

Syngnathidae (seahorses, seadragons and pipefish) suffer significant losses from non-tuberculous mycobacteria. However, they produce markedly different lesions in response to the disease compared to other teleost species, notably infrequent granuloma formation. This study evaluated 270 syngnathid fish, from which 92 were diagnosed with mycobacteriosis by histopathology, culture or both. Microscopic lesions variably consisted of random foci of coagulative necrosis in multiple organs, containing high numbers of free bacteria and large aggregates or sheets of macrophages with cytoplasm laden with acid-fast bacilli. Mycobacterial associated granulomas were identified in only six seahorses. Five fish had positive cultures with no observed microscopic changes. RNA-seq of the head kidney was performed to investigate the transcriptome of two infected and six non-infected lined seahorses Hippocampus erectus. Assembled and annotated putative transcripts serve to enrich the database for this species, as well as provide baseline data for understanding the pathogenesis of mycobacteriosis in seahorses. Putative components of the innate immune system (IL-1ß, IL-6, TNF, NOS, Toll-like receptor 1, MHC Class I, NF-κß, transforming growth factor beta, MyD88) were identified in the RNA-seq data set. However, a homolog for a key component in the TH1 adaptive immune response, interferon-gamma, was not identified and may underlie the unique pathologic presentation.

Gross, histological and ultrastructural morphology of the aglomerular kidney in the lined seahorse Hippocampus erectus.

Histologic evaluation of the renal system in the lined seahorse Hippocampus erectus reveals a cranial kidney with low to moderate cellularity, composed of a central dorsal aorta, endothelial lined capillary sinusoids, haematopoietic tissue, fine fibrovascular stroma, ganglia and no nephrons. In comparison, the caudal kidney is moderately to highly cellular with numerous highly convoluted epithelial lined tubules separated by interlacing haematopoietic tissue, no glomeruli, fine fibrovascular stroma, numerous capillary sinusoids, corpuscles of Stannius and clusters of endocrine cells adjacent to large calibre vessels. Ultrastructural evaluation of the renal tubules reveals minimal variability of the tubule epithelium throughout the length of the nephron and the majority of tubules are characterized by epithelial cells with few apical microvilli, elaborate basal membrane infolding, rare electron dense granules and abundant supporting collagenous matrix.

[Survival of patients with metastatic breast cancer according to pathological types of tumors]. / Cáncer de mama metastásico: Caracterización de una cohorte según subtipos.

BACKGROUND: The prognosis of breast cancer (BC) is in part determined by the stage at diagnosis and its pathological characteristics. AIM: To evaluate the association between survival of women with metastatic breast cancer and pathological features of the tumor. PATIENTS AND METHODS: We obtained clinical and pathological data from patients diagnosed with a metastatic BC between 1999 and 2013. The expression of estrogen (ER) and progesterone (PR) receptors and human epidermal growth factor receptor 2 (HER2) was determined by immunohistochemistry. Clinicopathological subtypes were defined as: Luminal A: ER or PR positive, HER2 negative, histological grade (HG) 1 or 2; Luminal B: ER or PR positive, HER2 negative or positive or HG 3; triple negative (TN): ER, PR and HER2 negative, independent of the HG, positive HER2: ER, PR negative and HER2 positive, independent of HG. We analyzed survival based on these subtypes. RESULTS: We identified 54 patients aged 24 to 85 years, with metastatic BC at diagnosis. Seventy five percent had luminal tumors; 19.6% HER2 positive and 7.8% were TN. In 61% of evaluable tumors, HG was classified as 3. The frequency of HER2 positive and high HG tumors was greater in these patients with metastatic BC than in a non-metastatic local BC cohort. Survival was higher among patients with Luminal tumors than in women with non-Luminal cancer (56.4 and 11.4 months, respectively, p = 0.04). CONCLUSIONS: Patients with metastatic BC at diagnosis often had HER2 positive tumors and high HG. As in other studies, ER positive tumors had a better survival.

[Characteristics and results of breast cancer in elderly females treated with curative intent]. / Cáncer de mama en mujeres adultas mayores, características clínicas e histopatológicas y resultados del tratamiento con intención curativa.

BACKGROUND: Aging is the main risk factor to acquire breast cancer (BC). Nevertheless BC in elderly patients is sub-represented in clinical trials. AIM: To describe the clinical characteristics and long term treatment results of localized BC in older women. MATERIAL AND METHODS: Review of medical records of 65 women aged 70 to 88 years, with localized BC, treated with surgery, postoperative radiotherapy or systemic therapy at a Clinical Hospital in Chile. RESULTS: The presence of symptoms or abnormal findings on physical examination were the main reasons for consulting in 65% of cases. Compared with tumors detected on physical examination, those detected using screening mammogram were smaller and were in stage 1 with a higher frequency (18 and 59% respectively, p < 0.01). The pathological type was luminal in 80% of cases. Overall survival was better for luminal pathological type. All patients completed radiotherapy without interruptions developing minor acute toxicity. The most common co-morbidity was high blood pressure occurring in 46% of patients. Thirteen percent of patients had three or more co-morbidities. After a median follow up of 7 years, 23 (35%) patients had died and the cause of death was BC in 43% of cases. Two patients died of lung cancer. No patient had a local breast relapse. CONCLUSIONS: Screening mammogram in older women detected smaller tumors and it was associated with a better survival. BC is the ultimate cause of death in approximately half of cases.

[Medullary thyroid cancer: review of 28 patients]. / Cáncer medular de tiroides: experiencia quirúrgica en 10 años.

BACKGROUND: Medullary thyroid cancer (MTC) represents approximately 5% of all thyroid cancers. Surgery is the only curative treatment, which includes total thyroidectomy and in most cases, neck dissection. AIM: To report our 10-year experience with surgical treatment of MTC. MATERIAL AND METHODS: Review of medical records and pathology reports of a university hospital. We retrieved data from 28 patients aged 47.2 ± 16 years (21 women) operated for a MTC treated between June 2002 and June 2012. RESULTS: In 20 patients, MTC was diagnosed in the preoperative period. Total thyroidectomy was performed in all cases and included a neck dissection in 24 patients. Median follow-up was 48 (2-120) months. Twenty-five patients (89.2%) achieved complete remission of the disease and three had disease recurrence. There were no deaths during the follow up. CONCLUSIONS: The diagnosis of MTC is mainly based on cytology. Total thyroidectomy with neck dissection is the treatment of choice. An early-stage diagnosis is associated with low rates of recurrence and absence of mortality.