Serbian Journal of Experimental and Clinical Research

PITUITARY HORMONE-PRODUCING CELLS AFTER ESTRADIOL APPLICATION IN THE RAT MODELS OF MENOPAUSE

Verica Milosevic — 2014-09-26

Female aging represents the biological process of structural and functional changes in endocrine cells and tissues, as well as in the pituitary hormone-producing cells. Besides the hypothalamic releasing hormones, estradiol plays the significant role in regulation of pituitary hormones synthesis/secretion, and it is still used for menopausal symptoms therapy. The effects of aging or ovariectomy, and synthetic estradiol application under these circumstances were evaluated in pituitary hormone-producing cells of female rats (animal models of menopause), i.e. observing the following cells: gonadotropes (FSH and LH), thyrotropes (TSH), somatotropes (GH), mammotropes (PRL) and corticotropes (ACTH). The cells were immunostained and histologically analyzed, while the ELISA method was used for hormonal analyses. Aging was found to cause diverse, commonly reductive changes regarding the volume, number and secretion of menopausal rat pituitary hormone-producing cells, except for the PRL cells that have significantly increased number and intesified secretion. After the treatment of middle-aged female rats with estradiol, absolute and relative pituitary weights are significantly increased in comparison with the control females. Histological parameters like the cell and volume density of PRL and ACTH cells are significantly increased compared to the control values. The mentioned parameters of FSH, LH and GH, and ocassionaly of TSH cells after estradiol treatment are significantly decreased in comparison with the controls. The corresponding hormone levels follow the changes in histological parameters. These data indicate that the estradiol application to menopausal females may specifically, in two directions, modify the histological characteristics and secretory activity of different pituitary-hormone producing cells.

Key words: female rat, middle-age, pituitary cells, estradiol.

UNFOLDED PROTEIN RESPONSE IS ACTIVATED IN HEART OF CATECHOLAMINERGIC POLYMORPHIC VENTRICULAR TACHYCARDIA (CPVT) MICE

Rigers Bakiu — 2014-09-26

ABSTRACT

Isoform 2 of calsequestrin (CSQ2), is the main calcium-binding protein of sarcoplasmic reticulum (SR), expressed both in cardiac and in skeletal muscles. CSQ2 acts as an SR calcium (Ca²⁺) sensor and regulates SR Ca²⁺ release via interactions with triadin, junctin, and the ryanodine receptor. Various mutations of the csq2 gene lead to altered Ca²⁺ release and contractile dysfunction contributing to the development of arrhythmias and sudden cardiac death in young individuals affected by CPVT. Transgenic mice carrying one of the identified CSQ2 point-mutations (R33Q) associated to CPVT were developed and a drastic reduction of the mutated protein was observed. Following a biomolecular approach several analysis were performed using different antibody treatments in order to identify when the reduction of CSQ2 begins, unveil the mechanism involved in the reduction of CSQ2 and verify if other proteins are affected by the presence of the mutated protein. This study results showed that mutated CSQ2 decreased soon after birth in conjunction with the decreased levels of other important proteins of the junctional SR, such as triadin (TD). Up-regulation of proteins associated to the unfolded protein response (UPR) was also observed and the pathway activated by UPR was the ATF6-dependent one. The R33Q mutation induced the decrease of CSQ2 by activation of the UPR and subsequently degradation through proteasome.

Keywords: Calsequestrin, CPVT, ERAD, UPR, Triadin, Chaperones.

BEHAVIORAL EFFECTS OF SHORT-TERM TOTAL FOOD RESTRICTION IN RATS

Dragica Selakovic — 2014-09-26

Food intake reduction can reduce indices of anxiety in rats. The aim of this study was to evaluate short-term (48 hours) total food restriction influence on behavioral characteristics, as well as motor coordination and balance of rats. 3 months old male Wistar albino rats (n=20) weighting between 350-400 g were divided into control group (food and water intake ad libitum) and experimental group - total food restriction 48 hours before testing. Behavioral studies were performed using: open field, elevated plus maze, Barnes maze, beam walking, evoked beam walking and linear locomotor test. Total distance moved, velocity, movement and frequency in centre zone of the open field were significantly higher in treated group. Cumulative duration in centre zone of the open field was not significantly increased in treated group. The number of entries into the open arms, the total time spent in open arms and total distance moved in the elevated plus maze were significantly increased with no change in velocity in food restricted animals. There was no effect of 48 hours total food restriction on the Barnes maze test parameters, as well as on parameters of linear locomotor test. The velocity recorded in beam walking test was not affected by food restriction, but the velocity recorded in evoked beam walking test significantly decreased in treated group. In summary, short-term total food restriction did not produce significant changes in physical performance of rats, but resulted in anxiolytic-like behavior accompanied with food-seeking behavior due to enhanced motivation to forage for food.

Key words: food restriction, behavior, anxiety, rat

The key role of leading emerging BRIC markets for the future of global health care

Mihajlo Jakovljevic — 2014-09-26

Acronym BRIC was created 2001 with the notion of marking the most promising and huge emerging markets outside the established postwar high income economies. Brazil, Russia, India and China’s nominal GDP growth rates continue to outpace the ones of Western Europe, North America and Japan before, during and after world macroeconomic crisis. This global phenomena will also heavily affect global demand and provision of health care services as well as other economy branches. The key cause for such change is occurrence of enormously massive middle class of citizens in these countries. Their health insurance coverage is extending together with package of services covered within such plans. Not less important is the overall growth of purchasing power followed by stronger affordability of vast portion of medical goods and services commonly paid out-of-pocket by ordinary citizens. Observing the changing landscape of global health care we should take into account steady slow economy growth rates of most mature, saturated markets. This also reflects to consumer demand for health services which remains strong in rich countries but true expansion of global market size happens actually elsewhere. This is acknowledged by all major market analysis agencies. They overtly recommend to multinational companies dealing with health care to focus on emerging markets and BRICs in particular if they are willing to survive. The key target to harvest long term profits and sustainable presence among pharmaceutical industry and medical equipment manufacturers worldwide shall remain emerging market investment for a long course of years to come.

KEY WORDS : BRICs, emerging markets, global, health care, demand, medical services

Xenobiotic Induced Model of Primary Biliary Cirrhosis

Aleksandar Arsenijevic — 2014-09-26

Primary biliary cirrhosis (PBC) is an autoimmune disease of the liver characterized by destruction of the intrahepatic bile ducts and the presence of antimitochondrial antibodies (AMAs). Several murine models of PBC with similar serological, biochemical, and histological features to human PBC have been developed in the recent years. These animal models enabled the investigation of the etiology and mechanisms involved in the pathogenesis of PBC. Immune response in PBC is directed towards E2 components of the 2-oxo-acid dehydrogenase family of enzymes located in mitochondria and immunodominant epitope is a lipoylated peptide sequence shared by these enzymes. Immunization of mice with 2-octynoic acid coupled to bovine serum albumin (2-OA-BSA), an antigen structurally related to the E2 subunit of the pyruvate dehydrogenase complex (PDC-E2), recapitulates the histologucal features of human PBC. This model of xenobiotic induced PBC is suitable for exploring the early events in PBC pathogenesis and for developing new therapeutics for PBC.

CUTANEOUS TOLERANCE OF SEA BUCKTHORN OIL EMULSION

Mihailo Kipic — 2014-09-26

Introduction: Sea buckthorn oil (Hippophae rhamnoides L.) is medically used both externally and internally, but the external application is unsutable due to its liquid, lipophylic and highly colored nature. These difficulties could be overbriged by formulation of semisolid emulsion with sea buckthorn oil. Previous research on this formulation showed that it has higher wound healing potential then sea buckthorn oil, possesing enhanced structure of liquid crystals, stability and suitability for topical use.

Aim: The aim of this investigation was to complete characterization of proposed emulsion, by testing its potential for causing skin irritation.

Methods: The emulsion was prepared by standard emulsifying techniques using a combination of surfactants that form an enhanced structure of liquid crystals. Amount of 40% of sea buckthorn oil was incorporated. Since results of previous research on this formulation show wound healing effect on an animal model, we also tested skin tolerance of it on animal skin model. Evaluation was done by Draize test which measures level of skin changes of rabbit skin.

Results: There was absence of edema or erythema type of irritation after 2h, 24h, 48h, 72h and 7 days of application of semisolid emuslion with sea buckthorn oil.

Conclusion: This study confirmed that combination of the proposed ingredients is adequate and the concentrations of surfactants used to form stable emulsion were appropriate also for skin application. The proposed formulation of emulsion with sea buckthorn oil exerted percutaneous tolerance and could be safely applied on the skin.

Key words: sea buckthorn oil, emulsion, topical use, irritation

ANTI – GBM RAPIDLY PROGRESSIVE GLOMERULONEPHRITIS (SYNDROMA GOODPASTURE) – CASE REPORT

Andreja M Figurek — 2014-09-26

ABSTRACT

Introduction. Goodpasture's syndrome is a severe illness caused by the formation of antibodies to the glomerular basement membrane and alveolus with consequential damages of renal and pulmonary function. With current therapy, long term survival is greater than 50%, and before it the mortality due to this disease was higher than 90%. Case report. In our patient, disease has began as dysuria, then continued as anemic syndrome, and at the end the end stage renal failure developed. Immunosuppressive therapy with pulse doses Methylprednisolone and Cyclophosphamide, has introduced this patient in the disease remission, but the permanent impairment of renal function still remained. Conclusion. Early diagnosis of Goodpasture's syndrom allows to preserve renal function and has a positive effect on the survival of patients. In patients who achieve remission of the disease, a kidney transplant can be considered. According to this, our patient is in processing for transplantation.

Key words: Syndroma Goodpasture; collagen α3 (IV) chain; anti-GBM antibodies; crescentic forms.