A non-functioning pancreatic neuroendocrine tumour: a case report

We present the diagnostic and therapeutic difficulties encountered in a patient with a clinically advanced pancreatic neuroendocrine tumour. The report concerns a 60-year-old female patient with the diagnosis of non-functioning pancreatic neuroendocrine tumour (NET G1) with liver, peripancreatic lymph node and mediastinal metastases. Due to the presence of advanced disease (inoperable pancreatic tumour, presence of multiple metastases) the patient was considered ineligible for surgical treatment on two occasions. Tissue samples for histopathology were collected during an exploratory laparotomy, which made it possible to establish the diagnosis. As somatostatin receptor scintigraphy was positive, the patient was started on somatostatin analogues and radionuclide therapy was initiated, resulting in satisfactory response in the form of complete remission of liver metastases and the decreased size of the primary tumour in the pancreas. The use of somatostatin analogues in the case of an inoperable neuroendocrine tumour which was assessed as clinically advanced, yet possessing a low proliferative potential, is a promising therapeutic option.     

Visualization of co-localization in Aβ42-administered neuroblastoma cells reveals lysosome damage and autophagosome accumulation related to cell death

Aβ42 [amyloid-β peptide-(1-42)] plays a central role in Alzheimer's disease and is known to have a detrimental effect on neuronal cell function and survival when assembled into an oligomeric form. In the present study we show that administration of freshly prepared Aβ42 oligomers to a neuroblastoma (SH-SY5Y) cell line results in a reduction in survival, and that Aβ42 enters the cells prior to cell death. Immunoconfocal and immunogold electron microscopy reveal the path of the Aβ42 with time through the endosomal system and shows that it accumulates in lysosomes. A 24?h incubation with Aβ results in cells that have damaged lysosomes showing signs of enzyme leakage, accumulate autophagic vacuoles and exhibit severely disrupted nuclei. Endogenous Aβ is evident in the cells and the results of the present study suggest that the addition of Aβ oligomers disrupts a crucial balance in Aβ conformation and concentration inside neuronal cells, resulting in catastrophic effects on cellular function and, ultimately, in cell death.     

Pyridobenzothiazole derivatives as new chemotype targeting the HCV NS5B polymerase

Hepatitis C virus (HCV) infection has been recognized as the major cause of liver failure that can lead to hepatocellular carcinoma. Among all the HCV proteins, NS5B polymerase represents a leading target for drug discovery strategies. Herein, we describe our initial research efforts towards the identification of new chemotypes as allosteric NS5B inhibitors. In particular, the design, synthesis, in vitro anti-NS5B and in cellulo anti-HCV evaluation of a series of 1-oxo-1H-pyrido[2,1-b][1,3]benzothiazole-4-carboxylate derivatives are reported. Some of the newly synthesized compounds showed an IC(50) ranging from 11 to 23 μM, and molecular modeling and biochemical studies suggested that the thumb domain could be the target site for this new class of NS5B inhibitors.     

Quality of life and marital sexual satisfaction in women with polycystic ovary syndrome

Polycystic ovary syndrome affects 5-10% of women in the developed world, making it the most common endocrine disorder among women of reproductive age. The symptoms typically associated with polycystic ovary syndrome: amenorrhea, oligomenorrhea, hirsutism, obesity, subfertility, anovulation and acne can lead to a significant reduction in female life quality.The aim of the study was to evaluate the effect of polycystic ovary syndrome on quality of life and marital sexual satisfaction. Fifty women with polycystic ovary syndrome were qualified to the study as the research group. The control group consisted of fourty healthy women. A specific questionnaire was used as a research tool in this study. It included the socio-demographic part, polycystic ovary syndrome's symptomatology and validated scales: Polish version of Short Form-36 Health Survey (SF-36) and Index of Sexual Satisfaction (ISS). The mean age of researched women was 28.9+/-5.6 years, and in the control group - 30.5+/-5.3 years (p>0.05). Quality of life parameters for women with polycystic ovary syndrome were lower than for the controls in the aspect of: general health (p<0.01), limitations due to physical health (p<0.05), limitations due to emotional problems (p<0.001), social functioning (p<0.01), energy/fatigue (p<0.001) and emotional wellbeing (p<0.01). Studied women showed worse marital sexual functioning (p<0.05). Marital sexual dysfunctions were diagnosed in 28.6% of women with polycystic ovary syndrome and in 10.5% of healthy women (p<0.05). Polycystic ovary syndrome decreases quality of life and marital sexual functioning among women. A negative effect of hirsutism severity on general well-being and marital sexual life is also observed.     

Granulosa cells regulate oocyte intracellular pH against acidosis in preantral follicles by multiple mechanisms

Mammalian oocytes grow within ovarian follicles in which the oocyte is coupled to surrounding granulosa cells by gap junctions. We report here that growing oocytes isolated from mouse preantral follicles are incapable of recovering from an experimentally induced acidosis, and that oocytes acquire the ability to manage acid loads by activating Na(+)/H(+) exchange during growth. By contrast, granulosa cells from similar preantral follicles possess substantial Na(+)/H(+) exchange capacity, which is attributable to the simultaneous action of two Na(+)/H(+) exchanger isoforms: NHE1 and NHE3. Granulosa cells were also found to possess a V-type H(+)-ATPase that drives partial acidosis recovery when Na(+)/H(+) exchange is inactivated. By monitoring intracellular pH (pH(i)) in small follicle-enclosed oocytes, we found that the oocyte has access to each of these acidosis-correcting activities, such that small follicle-enclosed oocytes readily recover from acidosis in a manner resembling granulosa cells. However, follicle-enclosed oocytes are unable to access these activities if gap-junction communication within the follicle is inhibited. Together, these experiments identify the NHE isoforms involved in regulating oocyte pH(i), indicate that gap junctions allow granulosa cells to exogenously regulate oocyte pH(i) against acidosis until the oocyte has acquired endogenous pH(i) regulation, and reveal that granulosa cells possess multiple mechanisms for carrying out this function.     

Insulinoma--diagnosis and treatment

Insulinomas are the most common functioning endocrine tumors of pancreas. Approximately 10% are multiple, less than 10% can be malignant and 5-10% associated with the MEN-1 syndrome. Insulinomas are the most common cause of hypoglycemia resulting from endogenous hyperinsulinism. The aim of this lecture is to present the up-to-date information concerning the prevalence, diagnosis and treatment of insulinoma.     

Infection of tobacco with different Pseudomonas syringae pathovars leads to distinct morphotypes of programmed cell death

Tobacco plants (Nicotiana tabacum cv. Xanthi-nc) infiltrated with either of two pathovars of Pseudomonas syringae- an avirulent strain of P. syringae pv. tabaci (Pst) or the non-host pathogen P. syringae pv. maculicola M2 (Psm) - developed a hypersensitive response (HR). There were considerable differences in HR phenotype, timing and sequence of cell dismantling between the two pathosystems. Following Psm infiltration, the first macroscopic signs were visible at 4.5 h post-infiltration (hpi). Simultaneously, increased plasma membrane permeability was observed, suggesting that the loss of cell membrane integrity initiates the macroscopic HR evoked by Psm. In contrast, after Pst treatment there was a distinct time lapse between the first signs of tissue collapse (9 hpi) and the occurrence of plasma membrane discontinuity (12 hpi). Ultrastructural studies of cells undergoing the HR triggered by Psm and Pst revealed distinct patterns of alterations in morphology of organelles. Moreover, while different forms of nuclear degeneration were observed in leaf zones infiltrated with Pst, we failed to detect any abnormalities in the nuclei of Psm-treated tissue. In addition, application of synthetic caspase inhibitors (Ac-DEVD-CHO, Ac-YVAD-CMK) abolished HR induced by Pst, but not Psm. Our observations suggest that different cell death mechanisms are executed in response to Psm and Pst. Interestingly, pre-inoculation with Pst, but not with Psm, induced a long-distance acquired resistance (LDAR) response, even though locally a typical set of defense responses, including acquired resistance, was activated locally in response to Psm. The failure of Psm to induce LDAR may be due to the rapid degeneration of bundle sheath cells resulting from Psm infection.     

Halorhabdus tiamatea: proteogenomics and glycosidase activity measurements identify the first cultivated euryarchaeon from a deep‐sea anoxic brine lake as potential polysaccharide degrader

Euryarchaea from the genus Halorhabdus have been found in hypersaline habitats worldwide, yet are represented by only two isolates: Halorhabdus utahensis AX‐2^T from the shallow Great Salt Lake of Utah, and Halorhabdus tiamatea SARL4B^T from the Shaban deep‐sea hypersaline anoxic lake (DHAL) in the Red Sea. We sequenced the H. tiamatea genome to elucidate its niche adaptations. Among sequenced archaea, H. tiamatea features the highest number of glycoside hydrolases, the majority of which were expressed in proteome experiments. Annotations and glycosidase activity measurements suggested an adaptation towards recalcitrant algal and plant‐derived hemicelluloses. Glycosidase activities were higher at 2% than at 0% or 5% oxygen, supporting a preference for low‐oxygen conditions. Likewise, proteomics indicated quinone‐mediated electron transport at 2% oxygen, but a notable stress response at 5% oxygen. Halorhabdus tiamatea furthermore encodes proteins characteristic for thermophiles and light‐dependent enzymes (e.g. bacteriorhodopsin), suggesting that H. tiamatea evolution was mostly not governed by a cold, dark, anoxic deep‐sea habitat. Using enrichment and metagenomics, we could demonstrate presence of similar glycoside hydrolase‐rich Halorhabdus members in the Mediterranean DHAL Medee, which supports that Halorhabdus species can occupy a distinct niche as polysaccharide degraders in hypersaline environments.     

Non-viral adeno-associated virus-based platform for stable expression of antibody combination therapeutics: Non-viral adeno-associated virus-based platform for stable expression of antibody combination therapeutics

Antibody combination therapeutics (ACTs) are polyvalent biopharmaceuticals that are uniquely suited for the control of complex diseases, including antibiotic resistant infectious diseases, autoimmune disorders and cancers. However, ACTs also represent a distinct manufacturing challenge because the independent manufacture and subsequent mixing of monoclonal antibodies quickly becomes cost prohibitive as more complex mixtures are envisioned. We have developed a virus-free recombinant protein expression platform based on adeno-associated viral (AAV) elements that is capable of rapid and consistent production of complex antibody mixtures in a single batch format. Using both multiplexed immunoassays and cation exchange (CIEX) chromatography, cell culture supernatants generated using our system were assessed for stability of expression and ratios of the component antibodies over time. Cultures expressing combinations of three to ten antibodies maintained consistent expression levels and stable ratios of component antibodies for at least 60 days. Cultures showed remarkable reproducibility following cell banking, and AAV-based cultures showed higher stability and productivity than non-AAV based cultures. Therefore, this non-viral AAV-based expression platform represents a predictable, reproducible, quick and cost effective method to manufacture or quickly produce for preclinical testing recombinant antibody combination therapies and other recombinant protein mixtures.