Ferritin-mediated iron detoxification promotes hypothermia survival in Caenorhabditis elegans and murine neurons.

How animals rewire cellular programs to survive cold is a fascinating problem with potential biomedical implications, ranging from emergency medicine to space travel. Studying a hibernation-like response in the free-living nematode Caenorhabditis elegans, we uncovered a regulatory axis that enhances the natural resistance of nematodes to severe cold. This axis involves conserved transcription factors, DAF-16/FoxO and PQM-1, which jointly promote cold survival by upregulating FTN-1, a protein related to mammalian ferritin heavy chain (FTH1). Moreover, we show that inducing expression of FTH1 also promotes cold survival of mammalian neurons, a cell type particularly sensitive to deterioration in hypothermia. Our findings in both animals and cells suggest that FTN-1/FTH1 facilitates cold survival by detoxifying ROS-generating iron species. We finally show that mimicking the effects of FTN-1/FTH1 with drugs protects neurons from cold-induced degeneration, opening a potential avenue to improved treatments of hypothermia.

Structural and functional relationship of mammalian and nematode ferritins.

Ferritin is a unique buffering protein in iron metabolism. By storing or releasing iron in a tightly controlled manner, it prevents the negative effects of free ferrous ions on biomolecules in all domains of life - from bacteria to mammals. This review focuses on the structural features and activity of the ferritin protein family with an emphasis on nematode ferritins and the similarities in their biological roles with mammalian ferritins. The conservative characteristic of the ferritin family across the species originates from the ferroxidase activity against redox-active iron. The antioxidative function of these proteins translates into their involvement in a wide range of important biological processes, e.g., aging, fat metabolism, immunity, anticancer activity, and antipathogenic activity. Moreover, disturbances in ferritin expression lead to severe iron-associated diseases. Research on the Caenorhabditis elegans model organism may allow us to better understand the wide spectrum of mechanisms involving ferritin activity.

The impact of in vitro cell culture duration on the maturation of human cardiomyocytes derived from induced pluripotent stem cells of myogenic origin.

Ischemic heart disease, also known as coronary artery disease (CAD), poses a challenge for regenerative medicine. iPSC technology might lead to a breakthrough due to the possibility of directed cell differentiation delivering a new powerful source of human autologous cardiomyocytes. One of the factors supporting proper cell maturation is in vitro culture duration. In this study, primary human skeletal muscle myoblasts were selected as a myogenic cell type reservoir for genetic iPSC reprogramming. Skeletal muscle myoblasts have similar ontogeny embryogenetic pathways (myoblasts vs. cardiomyocytes), and thus, a greater chance of myocardial development might be expected, with maintenance of acquired myogenic cardiac cell characteristics, from the differentiation process when iPSCs of myoblastoid origin are obtained. Analyses of cell morphological and structural changes, gene expression (cardiac markers), and functional tests (intracellular calcium transients) performed at two in vitro culture time points spanning the early stages of cardiac development (day 20 versus 40 of cell in vitro culture) confirmed the ability of the obtained myogenic cells to acquire adult features of differentiated cardiomyocytes. Prolonged 40-day iPSC-derived cardiomyocytes (iPSC-CMs) revealed progressive cellular hypertrophy; a better-developed contractile apparatus; expression of marker genes similar to human myocardial ventricular cells, including a statistically significant CX43 increase, an MHC isoform switch, and a troponin I isoform transition; more efficient intercellular calcium handling; and a stronger response to ß-adrenergic stimulation.

Techniques for the induction of human pluripotent stem cell differentiation towards cardiomyocytes.

The derivation of pluripotent stem cells from human embryos and the generation of induced pluripotent stem cells (iPSCs) from somatic cells opened a new chapter in studies on the regeneration of the post-infarction heart and regenerative medicine as a whole. Thus, protocols for obtaining iPSCs were enthusiastically adopted and widely used for further experiments on cardiac differentiation. iPSC-mediated cardiomyocytes (iPSC-CMs) under in vitro culture conditions are generated by simulating natural cardiomyogenesis and involve the wingless-type mouse mammary tumour virus integration site family (WNT), transforming growth factor beta (TGF-ß) and fibroblast growth factor (FGF) signalling pathways. New strategies have been proposed to take advantage of small chemical molecules, organic compounds and even electric or mechanical stimulation. There are three main approaches to support cardiac commitment in vitro: embryoid bodis (EBs), monolayer in vitro cultures and inductive co-cultures with visceral endoderm-like (END2) cells. In EB technique initial uniform size of pluripotent stem cell (PSC) colonies has a pivotal significance. Hence, some methods were designed to support cells aggregation. Another well-suited procedure is based on culturing cells in monolayer conditions in order to improve accessibility of growth factors and nutrients. Other distinct tactics are using visceral endoderm-like cells to culture them with PSCs due to secretion of procardiac cytokines. Finally, the appropriate purification of the obtained cardiomyocytes is required prior to their administration to a patient under the prospective cellular therapy strategy. This goal can be achieved using non-genetic methods, such as the application of surface markers and fluorescent dyes. Copyright © 2016 John Wiley & Sons, Ltd.

Techniques of Human Embryonic Stem Cell and Induced Pluripotent Stem Cell Derivation.

Developing procedures for the derivation of human pluripotent stem cells (PSCs) gave rise to novel pathways into regenerative medicine research. For many years, stem cells have attracted attention as a potentially unlimited cell source for cellular therapy in neurodegenerative disorders, cardiovascular diseases, and spinal cord injuries, for example. In these studies, adult stem cells were insufficient; therefore, many attempts were made to obtain PSCs by other means. This review discusses key issues concerning the techniques of pluripotent cell acquisition. Technical and ethical issues hindered the medical use of somatic cell nuclear transfer and embryonic stem cells. Therefore, induced PSCs (iPSCs) emerged as a powerful technique with great potential for clinical applications, patient-specific disease modelling and pharmaceutical studies. The replacement of viral vectors or the administration of analogous proteins or chemical compounds during cell reprogramming are modifications designed to reduce tumorigenesis risk and to augment the procedure efficiency. Intensified analysis of new PSC lines revealed other barriers to overcome, such as epigenetic memory, disparity between human and mouse pluripotency, and variable response to differentiation of some iPSC lines. Thus, multidimensional verification must be conducted to fulfil strict clinical-grade requirements. Nevertheless, the first clinical trials in patients with spinal cord injury and macular dystrophy were recently carried out with differentiated iPSCs, encouraging alternative strategies for potential autologous cellular therapies.

Simultaneous laparoscopic adrenalectomy and laparoscopic nephron-sparing surgery - new experience with port placement.

The aim of the study was to describe simultaneous laparoscopic adrenalectomy and laparoscopic nephron-sparing surgery, to discuss the details of a convenient laparoscopic approach and the way of port placement, as well as to present a review of the literature concerning combined laparoscopic procedures. A 72-year-old woman was admitted to our department because of a tumor of the right adrenal gland and a small tumor of the right kidney. The patient underwent simultaneous laparoscopic adrenalectomy and laparoscopic nephron-sparing surgery. The postoperative period was uncomplicated. The patient was discharged from the hospital on the 4(th) postoperative day. We believe that the proposed way of trocar placement would help to avoid a 'rollover' problem between the laparoscope and a Satinsky clamp or a 'crossing swords' problem between a Satinsky clamp and manipulators.

Monitoring respiration and cardiac activity using fiber Bragg grating-based sensor.

This paper shows the design of a fiber-based sensor for living activities in human body and the results of a laboratory evaluation carried out on it. The authors have developed a device that allows for monitoring the vibrations of human body evoked by living activities--breathing and cardiac rhythm. The device consists of a Bragg grating inscribed into a single mode optical fiber and operating on a wavelength of around 1550 nm. The fiber Bragg grating (FBG) is mounted inside a pneumatic cushion to be placed between the backrest of the seat and the back of the monitored person. Deformations of the cushion, involving deformations of the FBG, are proportional to the vibrations of the body leaning on the cushion. Laboratory studies have shown that the sensor allows for obtaining dynamic strains on the sensing FBG in the range of 50-124 µ strain caused by breathing and approximately 8.3 µstrain induced by heartbeat, which are fully measurable by today's FBG interrogation systems. The maximum relative measurement error of the presented sensor is 12%. The sensor's simple design enables it to be easily implemented in pilot's and driver's seats for monitoring the physiological condition of pilots and drivers.

Dismembered laparoscopic Anderson-Hynes pyeloplasty versus nondismembered laparoscopic Y-V pyeloplasty in the treatment of patients with primary ureteropelvic junction obstruction: a prospective study.

PURPOSE: An attempt has been made to prospectively compare the results of two laparoscopic pyeloplasties: Dismembered Anderson-Hynes (A-H) plasty and nondismembered Y-V plasty. Complications following the procedures have been studied as well. PATIENTS AND METHODS: Fifty patients with primary ureteropelvic junction obstruction (UPJO) were prospectively selected at random to undergo dismembered A-H (25 patients-even numbers) and nondismembered Y-V (25 patients-odd numbers) laparoscopic pyeloplasty. UPJO was diagnosed on the basis of ultrasonography, excretory urography, and diuretic renography (DR). The intensity of pain was assessed according to a visual analog pain scale (VAS). Success was defined by three factors taken collectively: 80% or greater pain relief in comparison with the preoperative VAS score, no obstruction on DR (decreasing renographic excretion curve, T(1/2) <12 min), and improved or stable differential renal function. The mean follow-up was 26.2 months for the A-H group and 26.6 months for Y-V group (P=0.865). RESULTS: Both groups were comparable in terms of preoperative data, except for the presence of the crossing vessel, which was more often observed in the Y-V group. No statistically significant differences between the studied groups were found in operative times, morbidity, and hospitalization length. The success rate in the A-H group was 95% and 86% in the Y-V group, the difference being not statistically significant. CONCLUSIONS: Laparoscopic A-H pyeloplasty achieved a higher success rate then Y-V pyeloplasty; however, the difference was not statistically significant.

Laparoscopic adrenalectomy - ten-year experience.

OBJECTIVES: The objective of the study is to summarize the authors' 10-year experience with laparoscopic adrenalectomy and to analyze the intra- and postoperative complications of the procedure. MATERIAL AND METHODS: The records of 80 patients who had undergone laparoscopic adrenalectomy from January 2002 to January 2012 were reviewed retrospectively. There were 51 female and 29 male patients. The average age was 52. In 33 cases the right adrenal gland was affected, in 47 it was the left adrenal gland. Nineteen operations were performed with the retroperitoneal approach, in 61 a transperitoneal access was used. The average size of the tumor was 5 cm. The diagnosis was based on ultrasonography (USG) and computed tomography (CT). The biochemical tests were performed in all cases to assess hormonal activity of the tumor. Pheochromocytoma was diagnosed in 16 cases, Cushing syndrome in 3 cases, and Conn syndrome in 4 cases. All other tumors were hormonally inactive. Six patients were operated on because of adrenal metastases - from renal carcinoma in five cases and from lung carcinoma in one case. RESULTS: There were three open conversions. The mean operative time was 158 minutes. The mean hospital stay was 5.5 days Blood transfusion was necessary in three patients. Postoperative complications were observed in 11 patients (13.7%). CONCLUSIONS: Laparoscopic adrenalectomy is a safe and effective procedure and should be considered the first - line treatment of benign adrenal masses. Our experience indicates that patients with adrenal metastases are suitable candidates for laparoscopic adrenalectomy, providing a skilled laparoscopic surgeon is involved in operation.