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Oseltamivir (Tamiflu) package labeling has a warning for neuropsychiatric adverse events (NPAE), most commonly in children and adolescents, especially males. There are several case reports of NPAE in adults treated with oseltamivir, but few document patients with preexisting neuropsychiatric conditions without additional contributing factors. This case report describes a 22-year-old male with a history of bipolar disorder, depression, and attention-deficit/hyperactivity disorder who had been stable on his medication regimen before experiencing sudden worsening of symptoms after the initiation of oseltamivir. The case adds to previous literature by strengthening the correlation between oseltamivir and a sudden increase in neuropsychiatric symptoms. Providers should be aware that oseltamivir may exacerbate symptoms of previously stable patients. Depending on the severity of neuropsychiatric effects, discontinuation of oseltamivir and symptom treatment with pharmacotherapy may be warranted.

The Venetic inscription from Kaštelir above Korte, bearing the siglum *Ts3, was incised onto a bowl sherd of grey depurated ware. It contains four graphemes in an unfragmented sequence that most likely reads vose, although the paleographic interpretation of the third grapheme remains somewhat problematic. The inscription very likely represents a personal name, probably in an abbreviated form, but one that has no exact match in the available corpus of Venetic inscriptions.

Mouse models of diet-induced type 2 diabetes mellitus provide powerful tools for studying the structural and physiological changes that are related to the disease progression. In this study, diabetic-like glucose dysregulation was induced in mice by feeding them a western diet, and light and transmission electron microscopy were used to study the ultrastructural changes in the pancreatic acinar cells. Acinar necrosis and vacuolization of the cytoplasm were the most prominent features. Furthermore, we observed intracellular and extracellular accumulation of lipid compounds in the form of lipid droplets, structural enlargement of the cisternae of the rough endoplasmic reticulum (RER), and altered mitochondrial morphology, with mitochondria lacking the typical organization of the inner membrane. Last, autophagic structures, i.e., autophagosomes, autolysosomes, and residual bodies, were abundant within the acinar cells of western diet-fed mice, and the autolysosomes contained lipids and material of varying electron density. While diets inducing obesity and type 2 diabetes are clearly associated with structural changes and dysfunction of the endocrine pancreas, we here demonstrate the strong effect of dietary intervention on the structure of acinar cells in the exocrine part of the organ before detectable changes in plasma amylase activity, which may help us better understand the development of non-alcoholic fatty pancreas disease and its association with endo- and exocrine dysfunction.

Tight control of beta cell stimulus-secretion coupling is crucial for maintaining homeostasis of energy-rich nutrients. While glucose serves as a primary regulator of this process, incretins augment beta cell function, partly by enhancing cytosolic [Ca2+] dynamics. However, the details of how precisely they affect beta cell recruitment during activation, their active time, and functional connectivity during plateau activity, and how they influence beta cell deactivation remain to be described. Performing functional multicellular Ca2+ imaging in acute mouse pancreas tissue slices enabled us to systematically assess the effects of the GLP-1 receptor agonist exendin-4 (Ex-4) simultaneously in many coupled beta cells with high resolution. In otherwise substimulatory glucose, Ex-4 was able to recruit approximately a quarter of beta cells into an active state. Costimulation with Ex-4 and stimulatory glucose shortened the activation delays and accelerated beta cell activation dynamics. More specifically, active time increased faster, and the time required to reach half-maximal activation was effectively halved in the presence of Ex-4. Moreover, the active time and regularity of [Ca2+]IC oscillations increased, especially during the first part of beta cell response. In contrast, subsequent addition of Ex-4 to already active cells did not significantly enhance beta cell activity. Network analyses further confirmed increased connectivity during activation and activity in the presence of Ex-4, with hub cell roles remaining rather stable in both control experiments and experiments with Ex-4. Interestingly, Ex-4 demonstrated a biphasic effect on deactivation, slightly prolonging beta cell activity at physiological concentrations and shortening deactivation delays at supraphysiological concentrations. In sum, costimulation by Ex-4 and glucose increases [Ca2+]IC during beta cell activation and activity, indicating that the effect of incretins may, to an important extent, be explained by enhanced [Ca2+]IC signals. During deactivation, previous incretin stimulation does not critically prolong cellular activity, which corroborates their low risk of hypoglycemia.

Instigated by relatively high cave-air CO2 concentrations in Modrič Cave (Croatia) recorded for the purpose of speleothem-based paleoclimate research, we established preliminary monitoring of radon (222Rn) concentrations within the cave for a 4.5-year period (2018–2022). As radioactive geogenic gas, radon, which often correlates with cave-air CO2 concentrations, presents a potential health hazard in cases of longer exposure time in high concentration conditions. Since the Modrič Cave is open to tourists and long-term scientific research has been performed within, a safety assessment for radon concentrations was essential. The integrated measurements of radon concentrations were performed by passive LR115 detectors that were exposed from three to six months at eight sites within the cave. Preliminary results showed seasonal variations of radon concentrations (0.08–13.6 kBq/m3) governed by the cave ventilation patterns, but superimposed on this, cave morphology and bedrock architecture control the radon variations on finer spatial scale. The 3-months average 222Rn concentration of up to 13.6 kBq/m3 during summer in one of the cave passages is among the highest measured seasonal averaged radon concentrations in Croatian caves, but maximum concentrations were even higher. Based on obtained results and calculations, potential negative health effects of radon exposure for cave visitors, guides and scientists were assessed and the results showed values of exposure to be below recommended levels. Calculated worst-case scenario for cave guides (most affected by radon and its progeny) revealed that they would receive dose slightly below the occupational dose limit of 20 mSv/y (19.0 ± 5.2 mSv/y) in the touristic part of the cave and significantly higher doses (34.1 ± 9.2 mSv/y) in the non-touristic part of the cave. To detect precise spatio-temporal radon concentration variations (up to diurnal scale) we recently established continuous radon measurements. This will enable detection of possibly health threatening short-term peaks in radon concentration and consequently further improve cave management.

Polysaccharide hydrogels and metal alloy nanoparticles have already found use in a range of biomedical applications. Nickel-copper nanoparticles (NiCu NPs) are particularly promising due to their tunable properties, such as ferromagnetism, biocompatibility, and antimicrobial activity. At the same time, polysaccharide hydrogels made of polymer mixtures such as alginate and methylcellulose with incorporated metal alloy nanoparticles are reported in the scientific literature. In view of this, in this work, NiCu NPs are combined with polysaccharide hydrogels and 3D printed to construct geometrically customizable dressings with tailorable properties for melanoma treatment. This novel combination exploits the intrinsic magnetic properties of NiCu NPs and the same time builds on their less known properties to improve the mechanic stability of 3D printed materials, both contributing to a previously not reported application as potent cytotoxic dressing against melanoma cells. The dressings were evaluated in terms of their physico-chemical characteristics, and their potential application, namely melanoma cell cytotoxicity. While all dressings exhibited similar degradation profiles regardless of composition, the addition of NiCu NPs had an effect on the hydrophilicity, swelling rates, and topographical properties of the dressings. Compression results showed that the presence of NPs increased the stiffness of the dressings, while the ultimate tensile strength was highest at 0.31 MPa for the dressings with 0.5 wt% NPs. We show that although the base formulation of the dressings is biocompatible with skin-derived cells, dressings loaded with NPs exhibit promising antimelanoma activity. Extracts obtained from dressings containing 0.5 wt% NPs reduced melanoma cell viability to 61% ± 11% and 40% ± 2% after 24 h and 72 h of soaking, respectively. Furthermore, extracts of dressings with 1 wt% NPs reduced melanoma cell viability to less than 15% within the first 24 h. By adjusting the NP content, the mechanical properties, surface roughness, and wettability can be tuned so that the dressings can be functionally customized. In addition, by using 3D printing as a fabrication process, the shape and composition of the dressings can be tailored to the patient's needs. The dressings also remained intact after soaking in simulated physiological solution for 14 days, indicating their suitability for long-term topical application.

Beta cells couple stimulation by glucose with insulin secretion and impairments in this coupling play a central role in diabetes mellitus. Cyclic adenosine monophosphate (cAMP) amplifies stimulus-secretion coupling via protein kinase A and guanine nucleotide exchange protein 2 (Epac2A). With the present research, we aimed to clarify the influence of cAMP-elevating diterpene forskolin on cytoplasmic calcium dynamics and intercellular network activity, which are two of the crucial elements of normal beta cell stimulus-secretion coupling, and the role of Epac2A under normal and stimulated conditions. To this end, we performed functional multicellular calcium imaging of beta cells in mouse pancreas tissue slices after stimulation with glucose and forskolin in wild-type and Epac2A knock-out mice. Forskolin evoked calcium signals in otherwise substimulatory glucose and beta cells from Epac2A knock-out mice displayed a faster activation. During the plateau phase, beta cells from Epac2A knock-out mice displayed a slightly higher active time in response to glucose compared with wild-type littermates, and stimulation with forskolin increased the active time via an increase in oscillation frequency and a decrease in oscillation duration in both Epac2A knock-out and wild-type mice. Functional network properties during stimulation with glucose did not differ in Epac2A knock-out mice, but the presence of Epac2A was crucial for the protective effect of stimulation with forskolin in preventing a decline in beta cell functional connectivity with time. Finally, stimulation with forskolin prolonged beta cell activity during deactivation, especially in Epac2A knock-out mice.

Patient management in the emergency department (ED) is evaluated by performance indicators, such as wait times (time to be seen by a physician), length of stay (LOS) and the number of diagnostic tests per patient. To improve the quality of care, dedicated emergency medicine (EM) specialists are employed to work in the ED. The aim of this study is to evaluate three performance indicators of patient management in the ED compared by specialty, internal medicine (IM) versus EM. Research was conducted in the ED of a large tertiary teaching hospital. A retrospective data analysis of the hospital information system was conducted for the period when only IM specialists were working as attendants, versus a period when two EM specialists joined the ED team. We calculated the percentage of patients seen within the recommended time per Australasian Triage system (AST) category and compared the average LOS and the average number of tests per patient, using data from June 2017 to January 2020. Means, standard deviation, standard error, 95% confidence interval were calculated, and the independent t-test was used to compare means. With the introduction of the EM specialists, the percentage of patients examined within the recommended time frame per AST category was higher. There was a significant reduction in LOS in the ED when comparing only IM specialists to IM specialists with two EM specialists (p

Alberto Fortis (1741-1803) was an Italian naturalist who had a wide range of scientific interests from linguistics and ethnology to geology, geography and hydrology. This paper presents an overview of hydrological features that Fortis recorded in two of his works: Saggio d'Osservazzioni sopra l'Isola di Cherso ed Osero (1771) and Viaggio in Dalmazia (1774). These works were a result of several Fortis' travels in the region and were noted in European scientific literature of that time. Fortis revealed to European public the eastern Adriatic coast, a then little-known part of Europe with its wide variety of natural features. Despite the scarce appearance of surface water in karst, numerous interpretations of hydrological features can be found in Fortis’ descriptions of the East Adriatic coast. He often shows a remarkable understanding of karst hydrology even though some of his ideas and explanations are dubious from the present point of view. Although Fortis is not considered a karstologist in the strict sense, his works contain elements of what will eventually become karstology.

Adrenaline inhibits insulin secretion from pancreatic beta cells to allow an organism to cover immediate energy needs by unlocking internal nutrient reserves. The stimulation of α2-adrenergic receptors on the plasma membrane of beta cells reduces their excitability and insulin secretion mostly through diminished cAMP production and downstream desensitization of late step(s) of exocytotic machinery to cytosolic Ca2+ concentration ([Ca2+]c). In most studies unphysiologically high adrenaline concentrations have been used to evaluate the role of adrenergic stimulation in pancreatic endocrine cells. Here we report the effect of physiological adrenaline levels on [Ca2+]c dynamics in beta cell collectives in mice pancreatic tissue slice preparation. We used confocal microscopy with a high spatial and temporal resolution to evaluate glucose-stimulated [Ca2+]c events and their sensitivity to adrenaline. We investigated glucose concentrations from 8-20 mM to assess the concentration of adrenaline that completely abolishes [Ca2+]c events. We show that 8 mM glucose stimulation of beta cell collectives is readily inhibited by the concentration of adrenaline available under physiological conditions, and that sequent stimulation with 12 mM glucose or forskolin in high nM range overrides this inhibition. Accordingly, 12 mM glucose stimulation required at least an order of magnitude higher adrenaline concentration above the physiological level to inhibit the activity. To conclude, higher glucose concentrations stimulate beta cell activity in a non-linear manner and beyond levels that could be inhibited with physiologically available plasma adrenaline concentration.

Stečki so srednjeveški kamniti nagrobniki, ki jih najdemo še vedno in situ v pokrajinah držav Zahodnega Balkana. Okoli 8% stečkov ima raznovrstne dekoracije v obliki nizkega reliefa, medtem, ko manj kot 400 nagrobnikov nosi tudi napise. Zaradi klimatskih vplivov vrezani napisi in ornamenti izginjajo s površine stečkov, zaradi česar jih je vse težje odkriti in prepoznati. V sklopu arheoloških raziskav stečkov na območju srednjeveške župe Dabar (današnji občini Berkovići in Bileća), je bil izveden pilotski projekt 3D skeniranja stečkov z ročnim skenerjem. Osem stečkov, dva z napisom in šest z dekoracijami, je bilo skeniranih v celoti, z namenom da bi lahko ovrednotili vidnost in prepoznavanje umetnostnih izrazov, ter samo tehniko detajlnega dokumentiranja materialne kulture. Članek predstavlja potek skeniranja in analizira rezultate.

Donepezil nasal delivery strategies are being continuously investigated for advancing therapy in Alzheimer’s disease. The aim of this study was to develop a chitosan-based, donepezil-loaded thermogelling formulation tailored to meet all the requirements for efficient nose-to-brain delivery. A statistical design of the experiments was implemented for the optimisation of the formulation and/or administration parameters, with regard to formulation viscosity, gelling and spray properties, as well as its targeted nasal deposition within the 3D-printed nasal cavity model. The optimised formulation was further characterised in terms of stability, in vitro release, in vitro biocompatibility and permeability (using Calu-3 cells), ex vivo mucoadhesion (using porcine nasal mucosa), and in vivo irritability (using slug mucosal irritation assay). The applied research design resulted in the development of a sprayable donepezil delivery platform characterised by instant gelation at 34 °C and olfactory deposition reaching a remarkably high 71.8% of the applied dose. The optimised formulation showed prolonged drug release (t1/2 about 90 min), mucoadhesive behaviour, and reversible permeation enhancement, with a 20-fold increase in adhesion and a 1.5-fold increase in the apparent permeability coefficient in relation to the corresponding donepezil solution. The slug mucosal irritation assay demonstrated an acceptable irritability profile, indicating its potential for safe nasal delivery. It can be concluded that the developed thermogelling formulation showed great promise as an efficient donepezil brain-targeted delivery system. Furthermore, the formulation is worth investigating in vivo for final feasibility confirmation.

Enzymes AKR1C regulate the action of oestrogens, androgens, and progesterone at the pre-receptor level and are also associated with chemo-resistance. The activities of these oestrone halides were investigated on recombinant AKR1C enzymes. The oestrone halides with halogen atoms at both C-2 and C-4 positions (13β-, 13α-methyl-17-keto halogen derivatives) were the most potent inhibitors of AKR1C1. The lowest IC50 values were for the 13α-epimers 2_2I,4Br and 2_2I,4Cl (IC50, 0.7 μM, 0.8 μM, respectively), both of which selectively inhibited the AKR1C1 isoform. The 13α-methyl-17-keto halogen derivatives 2_2Br and 2_4Cl were the most potent inhibitors of AKR1C2 (IC50, 1.5 μM, 1.8 μM, respectively), with high selectivity for the AKR1C2 isoform. Compound 1_2Cl,4Cl showed the best AKR1C3 inhibition, and it also inhibited AKR1C1 (Ki: AKR1C1, 0.69 μM; AKR1C3, 1.43 μM). These data show that halogenated derivatives of oestrone represent a new class of potent and selective AKR1C inhibitors as lead compounds for further optimisations.

Islets of Langerhans are multicellular microorgans located in the pancreas that play a central role in whole-body energy homeostasis. Through secretion of insulin and other hormones they regulate postprandial storage and interprandial usage of energy-rich nutrients. In these clusters of hormone-secreting endocrine cells, intricate cell-cell communication is essential for proper function. Electrical coupling between the insulin-secreting beta cells through gap junctions composed of connexin36 is particularly important, as it provides the required, most important, basis for coordinated responses of the beta cell population. The increasing evidence that gap-junctional communication and its modulation are vital to well-regulated secretion of insulin has stimulated immense interest in how subpopulations of heterogeneous beta cells are functionally arranged throughout the islets and how they mediate intercellular signals. In the last decade, several novel techniques have been proposed to assess cooperation between cells in islets, including the prosperous combination of multicellular imaging and network science. In the present contribution, we review recent advances related to the application of complex network approaches to uncover the functional connectivity patterns among cells within the islets. We first provide an accessible introduction to the basic principles of network theory, enumerating the measures characterizing the intercellular interactions and quantifying the functional integration and segregation of a multicellular system. Then we describe methodological approaches to construct functional beta cell networks, point out possible pitfalls, and specify the functional implications of beta cell network examinations. We continue by highlighting the recent findings obtained through advanced multicellular imaging techniques supported by network-based analyses, giving special emphasis to the current developments in both mouse and human islets, as well as outlining challenges offered by the multilayer network formalism in exploring the collective activity of islet cell populations. Finally, we emphasize that the combination of these imaging techniques and network-based analyses does not only represent an innovative concept that can be used to describe and interpret the physiology of islets, but also provides fertile ground for delineating normal from pathological function and for quantifying the changes in islet communication networks associated with the development of diabetes mellitus.

Introduction: The pandemic of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) presented a major challenge to health-care systems around the world. To ensure the continuity of hospital care for patients with cardiovascular diseases, the clinic has formulated the strategy for prevention of transmission of SARS-CoV-2 across the hospital environment. The purpose of this paper is to present the strategy for the prevention of transmission of SARS-CoV-2 in a healthcare facility, using Antigen Rapid Diagnostic Tests (Ag-RDTs). Methods: A description of the work from the hospital committee for the prevention and control of the SARS-CoV-2 epidemic is presented. The hospital has adopted the zero-case strategy. Each positive Ag-RDTs test was sent for confirmation by real-time Reverse Transcription Polymerase Chain Reaction (RT-PCR). Results: During the observed period, 6569 tests were performed at the Magdalena Clinic. Of these, a total of 6100 Ag-RDTs were performed, while 469 were recorded by RT-PCR test. Of these tests, a total of 181 tests showed a positive result, which is a share of 2.75% of all tests performed, of which 144 (2.19%) positive findings were detected among staff. In Ag-RDTs alone, a total of 86 positive people were detected, which is 1.41% of those tested on this type of test. Conclusion: The use of antigen rapid diagnostic tests is an effective and reliable method for the early detection of asymptomatic individuals infected with SARS-CoV-2 in a hospital setting. This strategy can significantly contribute to the insurance of uninterrupted healthcare, providing regular workflow processes, and the care of patients in the safest possible way.

Aggressive behaviour that follows acquired brain injury (ABI) is often hampering successful re-integration into the social life wherein causes and types vary. We focus on the consequences of ABI which are associated with the occurrence of aggressive behaviour and also hamper its treatment in the process of long-term rehabilitation. Specifically, we analyse changes in inhibitory control, reduced self-awareness, problems with accepting the situation and consequent emotional issues, and changes in personality and behaviour. In the process of long-term rehabilitation, we use an individually tailored approach, which aims to support and enable a person with ABI to develop more effective ways to cope with disabilities. As the outcomes in terms of progress are limited to specific situations, it is unlikely that aggressive behaviour would not repeat. We nevertheless find that knowledge of this phenomenon may reduce the frequency of aggressive behaviour and its impact on everyday functioning.

Although mice are a very instrumental model in islet beta cell research, possible phenotypic differences between strains and substrains are largely neglected in the scientific community. In this study, we show important phenotypic differences in beta cell responses to glucose between C57BL/6J, C57BL/6N, and NMRI mice, i.e., the three most commonly used strains. High-resolution multicellular confocal imaging of beta cells in acute pancreas tissue slices was used to measure and quantitatively compare the calcium dynamics in response to a wide range of glucose concentrations. Strain- and substrain-specific features were found in all three phases of beta cell responses to glucose: a shift in the dose-response curve characterizing the delay to activation and deactivation in response to stimulus onset and termination, respectively, and distinct concentration-encoding principles during the plateau phase in terms of frequency, duration, and active time changes with increasing glucose concentrations. Our results underline the significance of carefully choosing and reporting the strain to enable comparison and increase reproducibility, emphasize the importance of analyzing a number of different beta cell physiological parameters characterizing the response to glucose, and provide a valuable standard for future studies on beta cell calcium dynamics in health and disease in tissue slices.

Dressler syndrome is a rare type of secondary pericarditis that occurs after the injury of the heart by a myocardial infarction (MI), chest trauma, or heart surgery. The symptoms of Dressler syndrome usually present one to six weeks after the myocardial infarction with pleuritic chest pain, fever, dyspnoea, and ECG changes. It is estimated that only 0,1% of patients who have had acute myocardial infarction develop Dressler syndrome.

Hydronephrosis is a progressive dilatation of the renal calyces due to obstruction of urine outflow. If left untreated, the disease progresses to kidney failure and death.

Prostate cancer is the second most common malignant tumour in men. It usually presents with lower urinary tract symptoms and spreads to the regional lymph nodes. Metastasis to Virchow’s node (left supraclavicular lymphadenopathy) is rare and often associated with other types of cancer, such as gastric, lung and breast cancer. We present the case of a patient with enlarged left supraclavicular nodes as the first manifestation of disseminated adenocarcinoma of the prostate.

Endometrial cancer (EC) is the most common gynecological malignancy in resource-abundant countries. The majority of EC cases are estrogen dependent but the mechanisms of estrogen biosynthesis and oxidative metabolism and estrogen action are not completely understood. Here, we evaluated formation of estrogens in models of moderately and poorly differentiated EC: RL95-2 and KLE cells, respectively. Results revealed high expression of estrone-sulfate (E1-S) transporters (SLCO1A2, SLCO1B3, SLCO1C1, SLCO3A1, SLC10A6, SLC22A9), and increased E1-S uptake in KLE vs RL95-2 cells. In RL95-2 cells, higher levels of sulfatase and better metabolism of E1-S to E1 were confirmed compared to KLE cells. In KLE cells, disturbed balance in expression of HSD17B genes led to enhanced activation of E1 to E2, compared to RL95-2 cells. Additionally, increased CYP1B1 expression and down-regulation of genes encoding phase II metabolic enzymes: COMT, NQO1, NQO2, and GSTP1 suggested decreased detoxification of carcinogenic metabolites in KLE cells. Results indicate that in model cell lines of moderately and poorly differentiated EC, estrogens can be formed via the sulfatase pathway.

Spontaneous subthreshold activity in the central nervous system is fundamental to information processing and transmission, as it amplifies and optimizes sub-threshold signals, thereby improving action potential initiation and maintaining reliable firing. This form of spontaneous activity, which is frequently considered noise, is particularly important at auditory synapses where acoustic information is encoded by rapid and temporally precise firing rates. In contrast, when present in excess, this form of noise becomes detrimental to acoustic information as it contributes to the generation and maintenance of auditory disorders such as tinnitus. The most prominent contribution to subthreshold noise is spontaneous synaptic transmission (synaptic noise). Although numerous studies have examined the role of synaptic noise on single cell excitability, little is known about its pre-synaptic modulation owing in part to the difficulties of combining noise modulation with monitoring synaptic release. Here we study synaptic noise in the auditory brainstem dorsal cochlear nucleus (DCN) of mice and show that pharmacological potentiation of Kv3 K+ currents reduces the level of synaptic bombardment onto DCN principal fusiform cells. Using a transgenic mouse line (SyG37) expressing SyGCaMP2-mCherry, a calcium sensor that targets pre-synaptic terminals, we show that positive Kv3 K+ current modulation decreases calcium influx in a fifth of pre-synaptic boutons. Furthermore, while maintaining rapid and precise spike timing, positive Kv3 K+ current modulation increases the synchronization of local circuit neurons by reducing spontaneous activity. In conclusion, our study identifies a unique pre-synaptic mechanism which reduces synaptic noise at auditory synapses and contributes to the coherent activation of neurons in a local auditory brainstem circuit. This form of modulation highlights a new therapeutic target, namely the pre-synaptic bouton, for ameliorating the effects of hearing disorders which are dependent on aberrant spontaneous activity within the central auditory system.

NMDA receptors promote repolarization in pancreatic beta cells and thereby reduce glucose-stimulated insulin secretion. Therefore, NMDA receptors are a potential therapeutic target for diabetes. While the mechanism of NMDA receptor inhibition in beta cells is rather well understood at the molecular level, its possible effects on the collective cellular activity have not been addressed to date, even though proper insulin secretion patterns result from well-synchronized beta cell behavior. The latter is enabled by strong intercellular connectivity, which governs propagating calcium waves across the islets and makes the heterogeneous beta cell population work in synchrony. Since a disrupted collective activity is an important and possibly early contributor to impaired insulin secretion and glucose intolerance, it is of utmost importance to understand possible effects of NMDA receptor inhibition on beta cell functional connectivity. To address this issue, we combined confocal functional multicellular calcium imaging in mouse tissue slices with network science approaches. Our results revealed that NMDA receptor inhibition increases, synchronizes, and stabilizes beta cell activity without affecting the velocity or size of calcium waves. To explore intercellular interactions more precisely, we made use of the multilayer network formalism by regarding each calcium wave as an individual network layer, with weighted directed connections portraying the intercellular propagation. NMDA receptor inhibition stabilized both the role of wave initiators and the course of waves. The findings obtained with the experimental antagonist of NMDA receptors, MK-801, were additionally validated with dextrorphan, the active metabolite of the approved drug dextromethorphan, as well as with experiments on NMDA receptor KO mice. In sum, our results provide additional and new evidence for a possible role of NMDA receptor inhibition in treatment of type 2 diabetes and introduce the multilayer network paradigm as a general strategy to examine effects of drugs on connectivity in multicellular systems.

O-GlcNAcylation is an important post-translational and metabolic process in cells that must be carefully regulated. O-GlcNAc transferase (OGT) is ubiquitously present in cells and is the only enzyme that catalyzes the transfer of O-GlcNAc to proteins. OGT is a promising target in various pathologies such as cancer, immune system diseases, or nervous impairment. In our previous work we identified the 2-oxo-1,2-dihydroquinoline-4-carboxamide derivatives as promising compounds by a fragment-based drug design approach. Herein, we report the extension of this first series with several new fragments. As the most potent fragment, we identified 3b with an IC50 value of 116.0 μM. If compared with the most potent inhibitor of the first series, F20 (IC50 = 117.6 μM), we can conclude that the new fragments did not improve OGT inhibition remarkably. Therefore, F20 was used as the basis for the design of a series of compounds with the elongation toward the O-GlcNAc binding pocket as the free carboxylate allows easy conjugation. Compound 6b with an IC50 value of 144.5 μM showed the most potent OGT inhibition among the elongated compounds, but it loses inhibition potency when compared to the UDP mimetic F20. We therefore assume that the binding of the compounds in the O-GlcNAc binding pocket is likely not crucial for OGT inhibition. Furthermore, evaluation of the compounds with two different assays revealed that some inhibitors most likely interfere with the commercially available UDP-Glo™ glycosyltransferase assay, leading to false positive results. This observation calls for caution, when evaluating UDP mimetic as OGT inhibitors with the UDP-Glo™ glycosyltransferase assay, as misinterpretations can occur.

Beta cells within the pancreatic islets of Langerhans respond to stimulation with coherent oscillations of membrane potential and intracellular calcium concentration that presumably drive the pulsatile exocytosis of insulin. Their rhythmic activity is multimodal, resulting from networked feedback interactions of various oscillatory subsystems, such as the glycolytic, mitochondrial, and electrical/calcium components. How these oscillatory modules interact and affect the collective cellular activity, which is a prerequisite for proper hormone release, is incompletely understood. In the present work, we combined advanced confocal Ca2+ imaging in fresh mouse pancreas tissue slices with time series analysis and network science approaches to unveil the glucose-dependent characteristics of different oscillatory components on both the intra- and inter-cellular level. Our results reveal an interrelationship between the metabolically driven low-frequency component and the electrically driven high-frequency component, with the latter exhibiting the highest bursting rates around the peaks of the slow component and the lowest around the nadirs. Moreover, the activity, as well as the average synchronicity of the fast component, considerably increased with increasing stimulatory glucose concentration, whereas the stimulation level did not affect any of these parameters in the slow component domain. Remarkably, in both dynamical components, the average correlation decreased similarly with intercellular distance, which implies that intercellular communication affects the synchronicity of both types of oscillations. To explore the intra-islet synchronization patterns in more detail, we constructed functional connectivity maps. The subsequent comparison of network characteristics of different oscillatory components showed more locally clustered and segregated networks of fast oscillatory activity, while the slow oscillations were more global, resulting in several long-range connections and a more cohesive structure. Besides the structural differences, we found a relatively weak relationship between the fast and slow network layer, which suggests that different synchronization mechanisms shape the collective cellular activity in islets, a finding which has to be kept in mind in future studies employing different oscillations for constructing networks.

In postmenopausal women, estrogen levels exclusively depend on local formation from the steroid precursors dehydroepiandrosterone sulfate and estrone sulfate (E1-S). Reduced estrogen levels are associated with menopausal symptoms. To mitigate these symptoms, more women nowadays choose medicine of natural origin, e.g., Cimicifuga racemosa (CR), instead of hormone replacement therapy, which is associated with an increased risk of breast cancer, stroke, and pulmonary embolism. Although CR treatment is considered safe, little is known about its effects on healthy endometrial and ovarian tissue and hormone-dependent malignancies, e.g., endometrial and ovarian cancers that arise during menopause. The aim of our study was to examine the effects of CR on the expression of genes encoding E1-S transporters and estrogen-related enzymes in control and cancerous endometrial and ovarian cell lines. CR affected the expression of genes encoding E1-S transporters and estrogen-related enzymes only at very high concentrations, whereas no changes were observed at physiological concentrations of CR. This suggests that CR does not exert estrogenic effects in endometrial and ovarian tissues and probably does not affect postmenopausal women’s risks of endometrial or ovarian cancer or the outcomes of endometrial and ovarian cancer patients.

Reconstruction of the Quaternary environment is one of the main issues in the global endeavour of predicting future changes, to which the Mediterranean Sea is highly sensitive due to its latitude and landlocked position [...]

We present stable carbon (δ13C) and oxygen (δ18O) isotope records from two partially coeval speleothems from Manita peć Cave, Croatia. The cave is located close to the Adriatic coast (3.7 km) at an elevation of 570 m a.s.l. The site experienced competing Mediterranean and continental climate influences throughout the last glacial cycle and was situated close to the ice limit during the glacial phases. U-Th dating constrains the growth history from Marine Isotope Stage (MIS) 5 to MIS 3 and the transition from MIS 2 to MIS 1. 14C dating was used to estimate the age of the youngest part of one stalagmite found to be rich in detrital thorium and thus undatable by U-Th. On a millennial scale, δ18O variations partly mimic the Dansgaard–Oeschger interstadials recorded in Greenland ice cores (Greenland Interstadials, GI) from GI 22 to GI 13. We interpret our δ18O record as a proxy for variations in precipitation amount and/or moisture sources, and the δ13C record is interpreted as a proxy for changes in soil bioproductivity. The latter indicates a generally reduced vegetation cover towards MIS 3–MIS 4, with shifts of ~8‰ and approaching values close to those of the host rock. However, even during the coldest phases, when a periglacial setting and enhanced aridity sustained long-residence-time groundwater, carbonic-acid dissolution remains the driving force of the karstification processes. Speleothem morphology follows changes in environmental conditions and complements regional results of submerged speleothems findings. Specifically, narrow sections of light porous spelaean calcite precipitated during the glacial/stadial sea-level lowstands, while the warmer and wetter conditions were marked with compact calcite and hiatuses in submerged speleothems due to sea-level highstands. Presumably, the transformation of this littoral site to a continental one with somewhat higher amounts of orographic precipitation was a site-specific effect that masked regional environmental changes.

Pancreatic beta cells secrete insulin in response to stimulation with glucose and other nutrients, and impaired insulin secretion plays a central role in development of diabetes mellitus. Pharmacological management of diabetes includes various antidiabetic drugs, including incretins. The incretin hormones, glucagon-like peptide-1 and gastric inhibitory polypeptide, potentiate glucose-stimulated insulin secretion by binding to G protein-coupled receptors, resulting in stimulation of adenylate cyclase and production of the secondary messenger cAMP, which exerts its intracellular effects through activation of protein kinase A or the guanine nucleotide exchange protein 2A. The molecular mechanisms behind these two downstream signaling arms are still not fully elucidated and involve many steps in the stimulus–secretion coupling cascade, ranging from the proximal regulation of ion channel activity to the central Ca2+ signal and the most distal exocytosis. In addition to modifying intracellular coupling, the effect of cAMP on insulin secretion could also be at least partly explained by the impact on intercellular coupling. In this review, we systematically describe the possible roles of cAMP at these intra- and inter-cellular signaling nodes, keeping in mind the relevance for the whole organism and translation to humans.

Kaštelir above Korte near Izola is one of the best known prehistoric settlements in the Slovenian part of Istria. The site was first mentioned by Pietro Coppo in the 16th Century, at the beginning of 20th Century Carlo Marchesetti described the site as Il castelliere d‘Albuzzano presso Corte d‘Isola, however the first proper archaeological excavation took place in the 60’s and 70’s of 20th Century. With it the site was confirmed as a prehistoric hillfort of exceptional importance. Between 2008 and 2014 Kaštelir has been a main focus of the project “Kaštelir above Korte - Cultural and recreational park”, financially supported by the Municipality of Izola and conducted by research team from University of Primorska. In article we are presenting results of the archaeological test probing, done in 2014, which demonstrates that Kaštelir above Korte must be considered not only as a prehistoric settlement but also a place of a very early Roman settlement.

Even though stimulation and sensation seeking, which are typical for high-risk athletes, have been closely linked to high physical courage, little is known about physical courage of high-risk athletes. The purpose of this study was to examine physical courage among high- and non-risk athletes. The convenience sampling was used to reach both groups of the athletes. An anonymous online questionnaire was sent to the Faculty of Sport, several sport clubs and online sport forums in Slovenia. A total of 101 high-risk and 79 non-risk athletes filled out The Physical Courage Questionnaire and The Social Desirability Scale. In addition, important demographic characteristics were obtained. Results showed that high-risk athletes have a higher level of physical courage (F=6.304, p=0.013), and that male athletes are physically more courageous than female athletes (F=4.353, p=0.038). The interaction between gender and the risk of a given sport discipline had a significant effect on physical courage (F=4.936, p=0.028): in the non-risk athlete group female athletes reported being less physically courageous than male athletes, whereas high-risk female athletes scored similarly high on the physical courage scale as high-risk male athletes. Implications and limitations of the research study are discussed.

Abstract SNAP-25 is a protein of the core SNARE complex mediating stimulus-dependent release of insulin from pancreatic β cells. The protein exists as two alternatively spliced isoforms, SNAP-25a and SNAP-25b, differing in 9 out of 206 amino acids, yet their specific roles in pancreatic β cells remain unclear. We explored the effect of SNAP-25b-deficiency on glucose-stimulated insulin release in islets and found increased secretion both in vivo and in vitro. However, slow photo-release of caged Ca2+ in β cells within pancreatic slices showed no significant differences in Ca2+-sensitivity, amplitude or rate of exocytosis between SNAP-25b-deficient and wild-type littermates. Therefore, we next investigated if Ca2+ handling was affected in glucose-stimulated β cells using intracellular Ca2+-imaging and found premature activation and delayed termination of [Ca2+] i elevations. These findings were accompanied by less synchronized Ca2+-oscillations and hence more segregated functional β cell networks in SNAP-25b-deficient mice. Islet gross morphology and architecture were maintained in mutant mice, although sex specific compensatory changes were observed. Thus, our study proposes that SNAP-25b in pancreatic β cells, except for participating in the core SNARE complex, is necessary for accurate regulation of Ca2+-dynamics.

Endometrial cancer is the sixth most common cancer in women worldwide. It is associated with aberrant actions of steroid hormones, estrogens and progesterone, but also with enhanced inflammation and reduced cellular differentiation. Here, we show data on demographic and histopathological characteristics of 51 patients with endometrial cancer, together with data on correlations between the expression of 38 genes involved in estrogen and progesterone actions, inflammation and differentiation, and demographic characteristics. We also show data on changes in gene expression of these 38 genes according to histopathological and clinical characteristics of these patients. This article includes data referenced in the manuscript entitled »STAR and AKR1B10 are down-regulated in high-grade endometrial cancer by Sinreih et al. (in press) [1].

Nasal route of administration offers a unique opportunity of brain targeted drug delivery via olfactory and trigeminal pathway, providing effective CNS concentrations at lower doses and lower risk for adverse reactions compared to systemic drug administration. Therefore, it has been recently proposed as a route of choice for glucocorticoids to control neuroinflammation processes in patients with severe Covid-19. However, appropriate delivery systems tailored to enhance their efficacy yet need to emerge. In this work we present the development of sprayable brain targeting powder delivery platform of dexamethasone sodium phosphate (DSP). DSP-loaded microspheres, optimised employing Quality-by-Design approach, were blended with soluble inert carriers (mannitol or lactose monohydrate). Powder blends were characterized in terms of homogeneity, flow properties, sprayability, in vitro biocompatibility, permeability and mucoadhesion. Nasal deposition studies were performed using 3D printed nasal cavity model. Mannitol provided better powder blend flow properties compared to lactose. Microspheres blended with mannitol retained or enlarged their mucoadhesive properties and enhanced DSP permeability across epithelial model barrier. DSP dose fraction deposited in the olfactory region reached 17.0% revealing the potential of developed powder platform for targeted olfactory delivery. The observed impact of nasal cavity asymmetry highlighted the importance of individual approach when aiming olfactory region.

Diet-related immunometabolic-based diseases are associated with chronic inflammation in metabolic tissues, and infiltrated macrophages have been suggested as mediators for tissue- damaging inflammation. Growing evidence implicates Chenopodium quinoa and Salvia hispanica L. as important contributors to immunonutritional health. However, the functional roles of the immunonutritional protease inhibitors (PPIs) found in these crops on the macrophages’ metabolic and phenotypic adaptation remain to be elucidated. The salt soluble fraction of proteins was extracted and analyzed confirming the presence of 11S and 2S albumin. The S. hispanica in the bioaccessible fraction (BAF). Using human-like macrophage cells to deepen our understanding of the modulatory effects of this BAF, FACS analyses revealed their potential as TLR4 agonists, favoring increased phenotypic CD68/CD206 ratios. The results of mitochondrial stress tests showed that cells increased oxygen consumption rates and non-mitochondrial respiration, confirming negligible deleterious effects on mitochondrial function. At molecular-level, adaptation responses shed light on changes showing biological correlation with TLR4 signaling. The resulting immunometabolic effects triggered by PPIs can be a part of a tailored nutritional intervention strategy in immunometabolic-based diseases.

Beta cells in the pancreatic islets of Langerhans are precise biological sensors for glucose and play a central role in balancing the organism between catabolic and anabolic needs. A hallmark of the beta cell response to glucose are oscillatory changes of membrane potential that are tightly coupled with oscillatory changes in intracellular calcium concentration which, in turn, elicit oscillations of insulin secretion. Both membrane potential and calcium changes spread from one beta cell to the other in a wave-like manner. In order to assess the properties of the abovementioned responses to physiological and pathological stimuli, the main challenge remains how to effectively measure membrane potential and calcium changes at the same time with high spatial and temporal resolution, and also in as many cells as possible. To date, the most wide-spread approach has employed the electrophysiological patch-clamp method to monitor membrane potential changes. Inherently, this technique has many advantages, such as a direct contact with the cell and a high temporal resolution. However, it allows one to assess information from a single cell only. In some instances, this technique has been used in conjunction with CCD camera-based imaging, offering the opportunity to simultaneously monitor membrane potential and calcium changes, but not in the same cells and not with a reliable cellular or subcellular spatial resolution. Recently, a novel family of highly-sensitive membrane potential reporter dyes in combination with high temporal and spatial confocal calcium imaging allows for simultaneously detecting membrane potential and calcium changes in many cells at a time. Since the signals yielded from both types of reporter dyes are inherently noisy, we have developed complex methods of data denoising that permit for visualization and pixel-wise analysis of signals. Combining the experimental approach of high-resolution imaging with the advanced analysis of noisy data enables novel physiological insights and reassessment of current concepts in unprecedented detail.

This study presents high resolutionoxygen and carbon isotopic record of two U-Th dated stalagmites from the Eastern Adriatic caves. The stalagmites were collected from Mala špilja and Velika špilja caves situated on Mljet Island in the southern part of the Croatian Adriatic. Dripwater samples were collected from Medvjeđa špilja, Strašna peć, Špilja u Vrdolje, Kraljicina spilja, Velika špilja and Mala špilja caves. All caves formed in well-stratified Cretaceous limestones. The average value of deuterium excess of the dripwater is 9.3 ‰, indicating that the atmospheric conditions over the Atlantic Ocean have greater influence on the isotopic composition precipitation in the Adriatic region than the Eastern Mediterranean Sea. The longest isotopic record of the speleothems is of stalagmite MSM-1 from Mala špilja Cave dated from 119.2 ±3.3 ka to 5.6±0.6 ka, although deposition was not continuous. During the Holocene the speleothem grew between 7 ka and 4 ka. That interval is characterized by several δ18Oc and δ13Cc fluctuations reflecting changes in the environment. Superimposed on these fluctuations, there is an increasing trend in the 7.0−6.5 ka interval, evident in elevated δ18O values, which primarily refers the trend to drier conditions in the eastern Adriatic, which were occasionally interrupted by wet intervals. Humid conditions are particularly pronounced by low δ18Oc and δ13Cc values of speleothem MSM-1 between 7.3 and 6 ka, as a reflection of increased precipitation and lower temperatures. The transition to today's Mediterranean climate (Cs) occurred between 6 ka and 5 ka, while the driest conditions are recorded at ~4.7 − 4.2 ka. Comparisons between the eastern Adriatic δ18Oc and δ13Cc records of the speleothem from Mala špilja Cave (MSM-1) and Velika špilja Cave (Mljet Island) with Soreq Cave (Israel) and Corchia Cave (Italy) together with other proxies such as lake sediments demonstrate that the speleothems from the Eastern Adriatic caves record the local and the regional climatic changes. Key words: Holocene, speleothem; stable isotopes; Eastern Adriatic, Croatia. Rekonstrukcija okoljskih razmer v zgodnjem in poznem Holocenu na podlagi zapisa v sigi jam Mala Špilja in Velika Špilja (otok Mljet, Hrvaška) Raziskava temelji na visoko ločljivih zapisih kisikovih in ogljikovih izotopov, pridobljenih iz dveh stalagmitov, datiranih z uran-torijevo metodo. Kapnika sta bila vzorčena v jamah Mala špilja in Velika špilja na otoku Mljet, v južnem delu hrvaškega Jadrana. Vzorčena je bila tudi prenikla voda v jamah Medvjeđa špilja, Strašna peć, Špilja u Vrdolje, Kraljicina spilja, Velika špilja in Mala špilja. Vse omenjene jame so v plastovitem krednem apnencu. Povprečna vrednost presežka devterija v prenikli vodi je 9,3 ‰, kar kaže na prevladujoč podnebni vpliv Atlantskega oceana v primerjavi z vplivom vzhodnega Sredozemlja. Najdaljši izotopski zapis smo pridobili iz kapnika MSM-1 iz Male špilje, katerega starost je med 119,2 ±3,3 ka in 5,6±0,6 ka. Za kapnik je značilnih več prekinitev izločanja. V holocenu je kapnik rastel v obdobju med 7 ka in 4 ka. V tem obdobju je več izrazitih nihanj δ18Oc in δ13Cc, kar kaže na spremembe okolja. Med 7,0 in 6,5 ka je v zapisu trend naraščanja δ18O, kar kaže na relativno suhe razmere v vzhodnem Jadranu, ki so jih občasno prekinila vlažna obdobja. Med 7,3 in 6 ka je več obdobij z nizkim vrednostmi δ18Oc in δ13Cc v vzorcu MSM-1, kar kaže na veliko padavin in nizke temperature. Prehod v današnjo sredozemsko klimo (Cs) se je zgodil med 6 ka in 5 ka, najbolj suha obdobja smo zabeležili med 4,7 ka in 4,2 ka. Primerjava izotopskih zapisov δ18Oc on δ13Cc iz Male špilje (MSM-1) in Velike špilje (Mljet) z zapisi v jami Soreq (Izrael) in v sistemu Corchia (Italija) ter drugimi kazalci nekdanjih okolij, npr. jezerskimi sedimenti, kažejo, da sige vzhodne ga Jadrana hranijo pomembne zapise o lokalnih in regionalnih podnebnih spremembah. Ključne besede: Holocen, siga, stabilni izotopi, vzhodni Jadran, Hrvaška.

Speleothem-based research aimed to the reconstruction of the Quaternary environment in Dinaric karst has been conducted in several Croatian caves. As the interpretation of the palaeoenvironmental signal archived in speleothems relies on identifying and understanding processes between atmosphere and cave precipitates, especially hydrological behaviour, cave monitoring is crucial. Despite relatively simple sampling and monitoring protocols, series of problems and complications has arisen, caused by different natural and technical reasons. Extreme weather events such as enormous precipitation, prolonged drought or snow drifts, each in their own way can cause sample loss, while technical issues associated with equipment failures can lead to data loss. Even biological threats occurred, both man-induced and of natural origin. Although the later cannot be eliminated, as well as extreme weather, experience gained substantially helps to minimize negative effects. On the other hand, most of the technical issues are resolvable, and suggested solutions have been regularly implemented.

Endometrial cancer (EC) is the most common estrogen-dependent gynecological malignancy in the developed World. To investigate the local formation of estradiol (E2), we first measured the concentrations of the steroid precursor androstenedione (A-dione) and the most potent estrogen, E2, and we evaluated the metabolism of A-dione, estrone-sulfate (E1-S), and estrone (E1) in cancerous and adjacent control endometrium. Furthermore, we studied expression of the key genes for estradiol formation via the aromatase and sulfatase pathways. A-dione and E2 were detected in cancerous and adjacent control endometrium. In cancerous endometrium, A-dione was metabolized to testosterone, and no E2 was formed. Both, E1-S and E1 were metabolized to E2, with increased levels of E2 seen in cancerous tissue. There was no significant difference in expression of the key genes of the aromatase (CYP19A1) and the sulfatase (STS, HSD17B1, HSD17B2) pathways in cancerous endometrium compared to adjacent control tissue. The mRNA levels of CYP19A1 and HSD17B1 were low, and HSD17B14, which promotes inactivation of E2, was significantly down-regulated in cancerous endometrium, especially in patients with lymphovascular invasion. At the protein level, there were no differences in the levels of STS and HSD17B2 between cancerous and adjacent control tissue by Western blotting, and immunohistochemistry revealed intense staining for STS and HSD17B2, and weak staining for SULT1E1 and HSD17B1 in cancerous tissue. Our data demonstrate that in cancerous endometrium, E2 is formed from E1-S via the sulfatase pathway, and not from A-dione via the aromatase pathway.

Med zadnjo, to je pleistocensko-holocensko transgresijo, je naraščajoča morska gladina zalila velike dele dinarskega krasa. Ker porečja rek na vzhodni jadranski obali sestavljajo pretežno karbonatne kamnine, te reke prenašajo le okoli 20 % sedimentov v obliki suspenza, vse ostalo pa kot raztopljeni tovor. Zaradi tega je pod morsko gladino še vedno mogoče ugotavljati značilne kraške oblike, kot so škraplje, vrtače, polja, jame in brezna, kot tudi rečne doline in soteske. Poleg teh preprostih potopljenih oblik so zaradi dvigajoče se gladine nastale tudi nove: podmorski izviri imenovani vrulje, zaslanjeni obalni izviri in morska jezera. Najpomembnejši dokazi o nekdanjem kopnem okolju so kapniki v potopljenih jamah in lehnjak v potopljenih nekdanjih rekah. Te oboje je mogoče uporabiti za ugotavljanje nekdanje niæje morske gladine. During the last, Late Pleistocene-Holocene transgression, rising sea flooded a vast part of the Dinaric karst. Due to prevalence of carbonate rocks in the drainage area of most of the rivers on Eastern Adriatic coast, those rivers carry only approximately 20% of particulates as suspended matter and the rest is dissolved. Consequently, many typical karst features such as karrens, dolines, poljes, caves, pits and river valleys and canyons as well, presently under the sea, can still be recognized. Beside these simply drowned features, some new ones were formed by the sea level rise. Those are submarine springs, so called vruljas, brackish coastal springs and marine lakes. The most significant evidences of former subaerial conditions are speleothems in submerged caves and calc tufa deposits of drowned paleo rivers. Both of them could be used for determination of the former low sea level stands.

Z metodama 14C oziroma U-Th smo datirali 16 sig iz sedmih podmorskih jam v Jadranu. Dve sigi smo datirali z obema metodama. Primernost metod za datacijo sig je različna zaradi različnih okoljskih pogojev, ki so vladali med izločanjem sige oziroma po zalitju jame. Obe metodi sta zanesljivi na dobro ohranjenih sigah. U-Th metoda je slabša, ko so karbonati onesnaženi z detritičnim materialom oz. preraščeni s plastjo morskega izvora . Vendar metoda U-Th z uporabo MC ICPMS analize, kjer potrebujemo le 100-300mg vzorca (namesto 30 g pri metodi 14C), omogoča boljšo časovno ločljivost datacij, kar je pri datiranju sig zelo pomembno. Among the 16 speleothems that were collected from 7 submarine caves and pits for the purpose of 14C and U-Th dating and reconstructing sea-level changes, two speleothems were dated by both methods. Different environmental conditions during the speleothem deposition and after the submergence resulted with different appropriateness for speleothem dating by these techniques. Well preserved speleothems gave reliable results by both methods, while U-Th method showed disadvantage in the case of carbonates contaminated with detrital material, as well as in the case of carbonate from marine overgrowth that covers the speleothems. However, U-Th method using MC ICPMS technique which requires only 100-300 mg of sample per analysis (instead of ca. 30 g for 14C conventional method), offers better age resolution that is essential for speleothem dating.

This paper describes fractographic features observed in aerospace composites failed under tensile loads. Unidirectional Carbon Fibre Reinforced Plastic (UD CFRP) and Unidirectional Glass Fibre Reinforced Plastic (UD GFRP) composite specimens were fabricated and tested in tension. The morphology of fractured surfaces was studied at various locations to identify failure mechanism and characteristic fractographic features. CFRP composites displayed transverse crack propagation and the fracture surface showed three distinct regions, viz., crack origin, propagation and final failure. Significant variations in the fractographic features were noticed in crack propagation and final failure regions. Crack propagation region exhibited brittle fracture with chevron lines emanating from the crack origin. The entire crack propagation region exhibited radial marks on the individual fibre broken ends. On the other hand, the final fracture region revealed longitudinal matrix splitting and radial marks in majority of locations, and chop marks at some locations. The change in fracture mode in the final fracture was attributed to superimposition of bending loads. GFRP composites exhibited broom like fracture with extensive longitudinal splitting with radial marks present on individual fibre broken ends. Transverse fracture was observed at a few locations. These fracture features were analyzed and correlated with the loading conditions.

U-Th and 14C dating, and X-ray diffraction of the parts of 16 submerged speleothems taken from the depths of 1.5−41.5 m from 7 submarine caves and pits along the Eastern Adriatic coast, provided insight to the sea-level fluctuations during the last 220 ka and to the palaeogeographic changes caused by sea-level changes. Due to the climate changes, palaeoenvironmental settings also varied, but not so abrupt and intensive as in the rest of Europe. Namely, owing to the Alps and Dinarides acting as orographic barriers, Eastern Adriatic coast was the border region between periglacial Europe and temperate Mediterranean region. It was, also, refuge area for plant species from the north. This study showed that appropriate temperature, humidity and vegetation cover ensured favourable conditions for karstification and speleothems formation processes even during Last Glacial Maximum.

Rocky karstified coast, low tidal range, indented shoreline with numerous islands, diverse coastal biocenoses, abundant palaeontological, archaeological and historical evidence, relatively dense tide-gauge stations and developing GPS network – all these facts offer optimal prerequisite for the sea-level changes studies on the Croatian coast. Through various methodological approaches (geomorphological, biostratigraphical, archaeological/historical, mareographic, geodetic, radiometric), most of these issues have been used in order to reconstruct relative sea and land motions, yielding scattered and sometimes erroneous results. Unfortunately, some of them have been often uncritically used as the basis for the subsequent research. Only recently, with multidisciplinary approach, sequences of relative sea-level variations have been successfully revealed, but opportunities that eastern Adriatic coast offers will, undoubtedly, ensure relatively rapid research progress.

Stjenovita okršena obala, male amplitude morskih mijena, razvedena obala s brojnim otocima, različite priobalne biocenoze, mnoštvo paleontoloških, arheoloških i historijskih ostataka, dovoljan broj mareografskih postaja te razvoj GPS mreže – sve to nudi optimalne preduvjete za istraživanja promjena morske razine na hrvatskoj obali. Različitim metodološkim pristupima (geomorfološkim, biostratigrafskim, arheološkim, historijskim, mareografskim, geodetskim, radiometrijskim), većina ovih pogodnosti bila je korištena s ciljem rekonstruiranja relativnih pomaka morske razine i kopna. Međutim, proizašli rezultati bili su točkasti i nerijetko pogrešni, i takvi su često bili nekritički korišteni kao temelj novih istraživanja. Multidisciplinarnim pristupom tek su posljednjih godina djelomično rekonstruirane promjene relativne morske razine, a brojne pogodnosti istočne obale Jadrana nesumnjivo će omogućiti razmjerno brz napredak istraživanja.

Progesterone has a number of important functions throughout the human body. While the roles of progesterone are well known, the possible actions and implications of progesterone metabolites in different tissues remain to be determined. There is a growing body of evidence that these metabolites are not inactive, but can have significant biological effects, as anesthetics, anxiolytics and anticonvulsants. Furthermore, they can facilitate synthesis of myelin components in the peripheral nervous system, have effects on human pregnancy and onset of labour, and have a neuroprotective role. For a better understanding of the functions of progesterone metabolites, improved analytical methods are essential. We have developed a combined liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detection and quantification of progesterone and 16 progesterone metabolites that has femtomolar sensitivity and good reproducibility in a single chromatographic run. MS/MS analyses were performed in positive mode and under constant electrospray ionization conditions. To increase the sensitivity, all of the transitions were recorded using the Scheduled MRM algorithm. This LC-MS/MS method requires small sample volumes and minimal sample preparation, and there is no need for derivatization. Here, we show the application of this method for evaluation of progesterone metabolism in the HES endometrial cell line. In HES cells, the metabolism of progesterone proceeds mainly to (20S)-20-hydroxy-pregn-4-ene-3-one, (20S)-20-hydroxy-5α-pregnane-3-one and (20S)-5α-pregnane-3α,20-diol. The investigation of possible biological effects of these metabolites on the endometrium is currently undergoing.

Rab3a is a small GTPase of the Rab3 subfamily that acts during late stages of Ca²⁺-regulated exocytosis. Previous functional analysis in pituitary melanotrophs described Rab3a as a positive regulator of Ca²⁺-dependent exocytosis. However, the precise role of the Rab3a isoform on the kinetics and intracellular [Ca²⁺] sensitivity of regulated exocytosis, which may affect the availability of two major peptide hormones, α-melanocyte stimulating hormone (α-MSH) and β-endorphin in plasma, remain elusive. We employed Rab3a knock-out mice (Rab3a KO) to explore the secretory phenotype in melanotrophs from fresh pituitary tissue slices. High resolution capacitance measurements showed that Rab3a KO melanotrophs possessed impaired Ca²⁺-triggered secretory activity as compared to wild-type cells. The hampered secretion was associated with the absence of cAMP-guanine exchange factor II/ Epac2-dependent secretory component. This component has been attributed to high Ca²⁺-sensitive release-ready vesicles as determined by slow photo-release of caged Ca²⁺. Radioimmunoassay revealed that α-MSH, but not β-endorphin, was elevated in the plasma of Rab3a KO mice, indicating increased constitutive exocytosis of α-MSH. Increased constitutive secretion of α-MSH from incubated tissue slices was associated with reduced α-MSH cellular content in Rab3a-deficient pituitary cells. Viral re-expression of the Rab3a protein in vitro rescued the secretory phenotype of melanotrophs from Rab3a KO mice. In conclusion, we suggest that Rab3a deficiency promotes constitutive secretion and underlies selective impairment of Ca²⁺-dependent release of α-MSH.