Ultrastructural analysis of breast cancer patient-derived organoids

Background Breast cancer Patient Derived Organoids (PDO) have been demonstrated to be a reliable model to study cancer that promised to replace and reduce the use of animals in pre-clinical research. They displayed concordance with the tissue of origin, resuming its heterogenicity and representing a good platform to develop approaches of personalized medicines. Although obtain PDOs from mammary tumour, was a very challenging process, several ongoing studies evaluated them as a platform to study efficacy, sensitivity and specificity of new drugs and exploited them in personalized medicine. Despite tissue organization represented a crucial point to evaluate in a 3-dimensional model, since it could influence drug penetration, morphology of breast cancer PDOs has not been analysed yet. Here, we proposed a complete ultrastructural analysis of breast PDOs obtained from tumour and healthy tissues to evaluate how typical structures observed in mammary gland were resumed in this model. Methods 81 samples of mammary tissue (healthy or tumour) resulting from surgical resections have been processed to obtain PDO. The resulting PDOs embedded in matrigel drop have been processed for transmission electron microscopy and analysed. A comparison between ones from healthy and ones from cancerous tissue has been performed and PDOs derived from tumour tissue have been stratified according to their histological and molecular subtype. Result The morphological analysis performed on 81 PDO revealed an organized structure rich in Golgi, secretion granules and mitochondria, which was typical of cells with a strong secretory activity and active metabolism. The presence of desmosomes, inter and intracellular lumens and of microvilli and interdigitations signified a precise tissue-organization. Each PDO has been classified based on whether or not it possessed (i) peripheral ridges in mitochondria, (ii) intracellular lumens, (iii) intercellular lumens, (iv) micro-vesicles, (v) open desmosomes, (vi) cell debris, (vii) polylobed nuclei, (viii) lysosomes and (ix) secretion granules, in order to identify features coupled with the cancerous state or with a specific histological or molecular subtype. Conclusion Here we have demonstrated the suitability of breast cancer PDO as 3-dimensional model of mammary tissue. Besides, some structural features characterizing cancerous PDO have been observed, identifying the presence of distinctive traits.

Possible effect of thymoquinone on mast cell number and chymase, IL-4, IFN-γ expression in rat spleen

Thymoquinone (TQ) has been widely used in traditional medicine for the treatment of many diseases, to support the circulatory and immune system and to protect general health. Moreover, it is used as a nutritional supplement for preventive and therapeutic purposes in the respiratory, digestive and urinary systems. Secretion granules in the cytoplasm of mast cells contain primary mediators such as histamine, neutral proteases such as tryptase and chymase, and cytokines such as interleukin-4 (IL-4) and interferon-gamma (IFN-γ). 35 Sprague Dawley adult female rats were used as the study material. The rats were randomly assigned to five groups. First group: 1 ml/kg dose and second group: 2 ml/kg dose of TQ prepared at 1/1 (v/v) of ethanol and saline was intraperitoneally injected regularly in the rats daily for 42 days. Third group: 10 mg/kg dose and fourth group: 20 mg/kg dose of TQ was administered orally with the aid of a gavage probe. Fifth group was the control group in which no intervention was made. The lowest number of mast cells was detected in the group administered TQ at a dose of 20 mg/kg by oral gavage. It was determined that the numbers of mast cells in the control group and the group treated with TQ at a dose of 1 ml/kg intraperitoneally were close to each other. It was concluded that the increase or decrease between groups in the distribution of mast cells, chymase, IL-4, and IFN-γ cytokine expression may be partially effected in the spleen tissue by substances such as TQ.

Cytological Features of Mammary Analogue Secretory Carcinoma of the Parotid Gland in a 15-Year-Old Girl: A Case Report with Review of the Literature

Mammary analogue secretory carcinoma (MASC) is a recently recognized tumor of salivary glands characterized by the ETV6-NTRK3 fusion gene. This tumor is very rare in children and adolescents. We report a case of MASC in a 15-year-old girl, the fifth youngest case so far reported. The patient complained of a left infra-auricular mass that gradually enlarged for a year. Fine-needle aspiration cytology/imprint cytology showed individual tumor cells that had faintly eosinophilic granular cytoplasm with secretion granules sometimes seen adjacent to the tumor cells. These cytological features overlapped between those of zymogen granule-poor acinic cell carcinoma (AciCC) and MASC. In addition to the case report, we present a review of the related literature with a focus on the cytological features of MASC. The differential diagnostic clues are also discussed.

Ultrastructural profile of metapleural gland cells of the ant Atta laevigata (F. Smith, 1858) (Formicidae: Attini)

The metapleural gland is considered a synapomorphy of ants, and is characterized as a paired structure located at the two postlateral ends of the thorax and secretes substances capable of inhibiting and controlling fungi and bacteria in the fungus garden and inside the nest. This study was aimed at investigating if and which are the ultrastructural differences in the metapleural gland cells of workers (three castes) of Atta laevigata using transmission electron microscopy (TEM). This study revealed the presence of Golgi regions, rough endoplasmic reticulum (lamellar and vesicular shapes), smooth endoplasmic reticulum, mitochondria (elongated, round-shaped), vacuoles, secretion granules with different electron densities, and myelin figures in the cytoplasm of secretory cells, indicating that this gland produces substances composed of proteins, lipids, and polysaccharides (glycogen in major workers). Lipid droplets and secretion granules were found very near to the microvilli, especially in minor workers. The intracellular portion of canaliculi exhibited invaginations that increased the surface area and modified the secretion produced by the secretory cells. In the three castes examined, the gland exhibited a reservoir preceded by a collecting chamber, both lined by a simple squamous epithelium with a cuticular intima. Workers of the three castes of A. laevigata might be involved in the production of secretion mainly composed of proteins with antibiotic properties and, minor workers, may be responsible for producing a wider variety of secretions compared to median and major workers in the colony.

Feeding biology of carnivore and detritivore Mediterranean pycnogonids

The digestive system of sea spiders (Pycnogonida) presents peculiarities that have not been discussed in the context of their ecology or feeding behaviour. We investigated the digestive system of two Mediterranean species, a carnivorous speciesAmmothella longipesand a detritivorousEndeis spinosa, with special focus on its correlation with behavioural feeding habits. The midgut and hindgut sections did not present significant differences between the two species, but major differences were observed in the foregut, reflecting concordance to their diet and their feeding behaviour. Jaws, setose lips, the structure of the pharyngeal filter and musculature of the proboscis are the main differential elements when comparing feeding habits ofA. longipesandE. spinosa. These elements are responsible for the reduction of the food pulp down to subcellular size. The digestion process observed in the species studied agrees with that observed in other pycnogonid lineages, but differs from most marine arthropods mainly because of the absence of midgut gland cells and the presence of a unique multifunctional type of midgut epithelial cell. Epithelial digestive cells are present in a small ‘resting’ form during starvation periods. During digestion, secretion granules possibly containing zymogen move to their apical border to be secreted to the midgut lumen, secondary lysosomes are formed and intracellular digestion occurs within them. Residual bodies are formed within the epithelial cell and released to the midgut lumen to be transported towards the hindgut. The characteristics of the digestive process of the pycnogonids studied seem to reflect a plesiomorphic state in arthropods.

Redescription of Capitella capitata (Fabricius) from West Greenland and designation of a neotype (Polychaeta, Capitellidae)

Capitella capitata (Fabricius, 1780), the type species of the genus, was originally described from the SW coast of Greenland; however, current definitions of the species are largely based on Eisig (1887), who described specimens from Naples on the Mediterranean coast. Eisig’s excellent account of the morphology of a sexually dimorphic species has subsequently been applied to specimens collected from worldwide locations. C. capitata was considered cosmopolitan in its distribution until J.P. Grassle and others identified numerous sibling species in laboratory culture. A large collection of Capitella from locations along the W, SW, and SE coasts of Greenland was provided by the Zoological Museum, University of Copenhagen (ZMUC). The collections included about 680 specimens with distinctive prostomial, thoracic, and setal morphology that differs from the typical depiction of the species in most faunal accounts. A neotype was selected from an excellent collection of approximately 250 specimens from Mârmorilik on the Affarlikassâ Fjord on the inner Uummannaq Fjord system, north of Disko Bay. All available collections from the ZMUC are believed to belong to the same species, which is here considered to be Arctic and subarctic in distribution. Additional material from the Alaskan Arctic supports this hypothesis. The Greenland species has a short, cup-shaped, wider-than-long prostomium joined to a narrow peristomial ring that is retracted slightly into setiger 1. In most specimens, the first four thoracic setigers gradually increase in size, with setiger 4 being the largest thoracic segment, and then decrease over the next five setigers, with setiger 9 being smaller than subsequent anterior abdominal segments. The thoracic region is thus markedly distinct visually from the abdominal region. Setigers 8–9 typically lack fascicles of notosetae, but 80% of the specimens examined have genital spines and most have copulatory organs that are secretory in nature: they contain numerous small secretion granules but no sperm. Many of these specimens have eggs and larvae within their tubes, suggesting functional hermaphroditism. The present redescription of C. capitata serves to establish a baseline against which the many known but undescribed species can be compared. Previous accounts of C. capitata based on the Eisig concept and largely from boreal and temperate localities should be reevaluated.

Multimerin 1 binds factor V and activated factor V with high affinity and inhibits thrombin generation

SummaryMultimerin 1 (MMRN1) is a polymeric, factorV (FV) binding protein that is stored in platelet and endothelial cell secretion granules but is undetectable in normal plasma. In human platelet α-granules, FV is stored complexed to MMRN1, predominantly by noncovalent binding interactions. The FV binding site for MMRN1 is located in the light chain, where it overlaps the C1 and C2 domain membrane binding sites essential for activated FV (FVa) procoagulant function. Surface plasmon resonance (SPR), circular dichroism (CD) and thrombin generation assays were used to study the binding of FV and FVa to MMRN1, and the functional consequences. FV and FVa bound MMRN1 with high affinities (KD:2 and 7 nM, respectively). FV dissociated more slowly from MMRN1 than FVa in SPR experiments, and CD analyses suggested greater conformational changes in mixtures of FV and MMRN1 than in mixtures of FV and MMRN1. SPR analyses indicated that soluble phosphatidylserine (1,2-Dicaproylsn-glycero-3-phospho-L-serine) competitively inhibited both FV-MMRN1 and FVa-MMRN1 binding. Furthermore, exogenous MMRN1 delayed and reduced thrombin generation by plasma and platelets, and it reduced thrombin generation by preformed FVa. Exogenous MMRN1 also delayed FV activation, triggered by adding tissue factor to plasma, or by adding purified thrombin or factor Xa to purified FV. The high affinity binding of FV to MMRN1 may facilitate the costorage of the two proteins in platelet α-granules. As a consequence, MMRN1 release during platelet activation may limit platelet dependent thrombin generation in vivo.