Increasing adherence to the Mediterranean diet and lifestyle is associated with reduced fecal calprotectin and intra-individual changes in microbial composition of healthy subjects.

The Mediterranean diet (MED) is associated with the modification of gut microbial composition. In this pilot study, we investigate the feasibility of a microbiota-targeted MED-based lifestyle intervention in healthy subjects. MED intervention integrating dietary counseling, a supporting mobile application, and daily physical activity measurement using step trackers was prospectively applied for 4 weeks. Blood and fecal samples were collected at baseline, after the 4-week intervention, and at 6 and 12 months. Blood counts, inflammatory markers, microbial and eukaryotic composition were analyzed. Dietary adherence was assessed using daily questionnaires. All 20 healthy participants (females 65%, median age 37), completed the 4-week intervention. Adherence to MED increased from 15.6 ± 4.1 (baseline) to 23.2 ± 3.6 points (4 weeks), p < .01, reflected by increased dietary fiber and decreased saturated fat intake (both p < .05). MED intervention modestly reduced fecal calprotectin, white blood cell, neutrophil, and lymphocyte counts, within the normal ranges (P < .05). Levels of butyrate producers including Faecalibacterium and Lachnospira were positively correlated with adherence to MED and the number of daily steps. Bacterial composition was associated with plant-based food intake, while fungal composition with animal-based food as well as olive oil and sweets. Increasing adherence to MED correlated with increased absolute abundances of multiple beneficial gut symbionts. Therefore, increasing adherence to MED is associated with reduction of fecal calprotectin and beneficial microbial alterations in healthy subjects. Microbiota targeted lifestyle interventions may be used to modify the intestinal ecosystem with potential implications for microbiome-mediated diseases.

The relation between gain in cognition during rehabilitation on functional outcome among hip fracture adult patients with and without pre- hip fracture dementia.

The purpose of the present study was focused on the relationship between change in cognition and the functional outcome during rehabilitation in demented and non-demented adult hip fracture patients. We studied seventy consecutive adult patients with hip fracture admitted to our rehabilitation wards. Functional outcome was assessed by the Functional Independence Measure (FIM). The gain in cognition during the rehabilitation process was measured by the difference in Mini Mental State Examination scores at admission and discharge. Data was analyzed by t-test, chi square-test and linear regression. Patients without dementia presented and discharged from the rehabilitation ward with statistically significant higher total, motor, and gain functional independence measure scores compared to patients with dementia. In a multiple regression analyses, gain in Mini Mental State scores examination were not independently associated with higher total and motor functional independence measure scores at discharge (beta = 0.086, p = 0.194; beta = 0.077, p = 0.309, respectively). Our findings suggest that there is no association between functional outcome and cognitive gain at the end of the rehabilitation process among adult hip fracture patients with and without dementia. However hip fracture adult patients with dementia should not be deprived of a post-acute rehabilitation.

Disruption of the actin cytoskeleton leads to inhibition of mitogen-induced cyclin E expression, Cdk2 phosphorylation, and nuclear accumulation of the retinoblastoma protein-related p107 protein.

The actin cytoskeleton has been found to be required for mitogen-stimulated cells to passage through the cell cycle checkpoint. Here we show that selective disruption of the actin cytoskeleton by dihydrocytochalasin B (H(2)CB) blocked the mitogenic effect in normal Swiss 3T3 cells, leading to cell cycle arrest at mid to late G(1) phase. Cells treated with H(2)CB remain tightly attached to the substratum and respond to mitogen-induced MAP kinase activation. Upon cytoskeleton disruption, however, growth factors fail to induce hyperphosphorylation of the retinoblastoma protein (pRb) and the pRb-related p107. While cyclin D1 induction and cdk4-associated kinase activity are not affected, induction of cyclin E expression and activation of cyclin E-cdk2 complexes are greatly inhibited in growth-stimulated cells treated with H(2)CB. The inhibition of cyclin E expression appears to be mediated at least in part at the RNA level and the inhibition of cdk2 kinase activity is also attributed to the decrease in cdk2 phosphorylation and proper subcellular localization. The expression patterns of cdk inhibitors p21 and p27 are similar in both untreated and H(2)CB-treated cells upon serum stimulation. In addition, the changes in subcellular localization of pRb and p107 appear to be linked to their phosphorylation states and disruption of normal actin structure affects nuclear migration of p107 during G(1)-to-S progression. Taken together, our results suggest that the actin cytoskeleton-dependent G(1) arrest is linked to the cyclin-cdk pathway. We hypothesize that normal actin structure may be important for proper localization of certain G(1) regulators, consequently modulating specific cyclin and kinase expression.

Dynamic and consequential consistency of choices between paths of decision trees.

The generally prescribed procedure for choosing a decision strategy from a decision tree employs a backward induction analysis that entails 3 fundamental consistency principles: dynamic, consequential, and strategic. The first requires the decision maker to follow through on plans to the end, the second requires the decision maker to focus solely on future events and final consequences given the current state of events, and the third is the conjunction of the first 2. Five experiments were reported to test these principles using different subject populations, different procedures for estimating consistency, and different factors for manipulating the attractiveness of the gamble at the final stage of the tree. The main findings were that strategic and dynamic consistency principles were violated at rates that exceeded choice inconsistency.

Changing plans: dynamic inconsistency and the effect of experience on the reference point.

This article presents a new test of a principle of decision making called dynamic consistency. This principle was tested in an experiment in which participants were asked to make decisions about a second gamble within a sequence of two gambles. Participants were first asked to make a planned choice about the second gamble. The planned choice was made before the first gamble was played and was conditioned on the anticipated outcomes of the first gamble. After the first gamble was played, the same participants were asked to make a final choice about the second gamble, conditioned on the experienced outcome of the first gamble. The results showed that participants' final choices were frequently inconsistent with their plans, even when the anticipated and experienced outcomes were identical. These inconsistencies occurred in a systematic direction. Experiencing an anticipated gain resulted with a change toward risk aversion, and experiencing an anticipated loss resulted in a change toward risk seeking. These results are explained in terms of the effect of actual experience on the reference point used for the evaluation of the decision problem.

Accidents and Decision Making under Uncertainty: A Comparison of Four Models.

Heinrich's (1931) classical study implies that most industrial accidents can be characterized as a probabilistic result of human error. The present research quantifies Heinrich's observation and compares four descriptive models of decision making in the abstracted setting. The suggested quantification utilizes signal detection theory (Green & Swets, 1966). It shows that Heinrich's observation can be described as a probabilistic signal detection task. In a controlled experiment, 90 decision makers participated in 600 trials of six safety games. Each safety game was a numerical example of the probabilistic SDT abstraction of Heinrich's proposition. Three games were designed under a frame of gain to represent perception of safe choice as costless, while the other three were designed under a frame of loss to represent perception of safe choice as costly. Probabilistic penalty for Miss was given at three different levels (1, .5, .1). The results showed that decisions tended initially to be risky and that experience led to safer behavior. As the probability of being penalized was lowered decisions became riskier and the learning process was impaired. The results support the cutoff reinforcement learning model suggested by Erev et al. (1995). The hill-climbing learning model (Busemeyer & Myung, 1992) was partially supported. Theoretical and practical implications are discussed. Copyright 1998 Academic Press.

[Dependence of the process leading to phosphorylation of retinoblastoma protein (pRb) on the status of the actin cytoskeleton]. / Zavisimost' protsessa, vedushchego k fosforilirovaniiu belka retinoblastomy (pRb), ot sostaiianiia aktinovogo tsitoskeleta.

It has been confirmed that the main actin-dependent period of G1 phase of the cell cycle is the middle of G1. As the critical points in G1 phase are associated with the synthesis of cycling D and E and with the formation of their active complexes with cyclin-dependent kinases (Cdk's), a study of a possible influence of actin filaments on these processes was performed. The activity of G1 kinases was estimated by the degree of phosphorylation of their specific substrate-retinoblastoma protein (pRb). Immunoblot analysis with specific antibodies to pRb revealed hypophosphorylated from of Rb in lysates of resting Swiss 3T3 cells and the appearance of hyperphosphorylated form after 12 h of EGF and serum stimulation. In was shown that actin filaments distruction by H2CB led to a decrease in hyper phosphorylated form appearance, depending on the depth of resting state of the cells and efficiency of their stimulation by growth factors. Thus, these data may suggest the involvement of actin cytoskeleton in functioning of the transcription chain cyclin/Cdk-R1-E2F.

[The role of the actin cytoskeleton in cell transition through the middle of the G1 phase of the mitotic cycle]. / Rol' aktinovogo tsitoskeleta v prokhozhdenii kletkami serediny fazy G1 mitoticheskogo tsikla.

The influence of dihydrocylochalasin B (H2CB), which selectively disrupts the actin cytoskeleton structure, on G1 phase progression after stimulation of quiescent Swiss 3T3 cells by epidermal growth factor (EGF) was investigated. H2CB was added to the culture medium (10 mg/ml) for different periods of time after cell cycle induction by EGF (10 ng/ml). Efficiency of mitogenic stimulation was estimated by 14C-thymidine uptake 21-23 h after EGF addition. It is shown that the main actin-mediated step is in the middle of G1, from 8 to 12 h after stimulation. Disorganization of actin cytoskeleton structure only during this time led to complete and irreversible block of cell entry into the S phase. On the contrary, the same effect of H2CB during the early period of G1 (first 6 h) led to the increase in 14C-thymidine incorporation in DNA. The specificity of actin-dependent period was proven in experiments with another inhibitor of cell proliferation--dancylcadaverine, whose effect was revealed at the earlier time--4-6 h after cell stimulation. Inhibition of protein synthesis during actin-dependent period (8-12 h) led to the same block of cell progression through G1 as it was seen after actin structure disruption. These data suggest that actin-dependent block is associated with the appearance and functioning of such specific regulators of G1 as cyclins (D, E) and their complexes with Cdk's (G1 kinases) which phosphorylate Rb protein (p1 10) associated with transcription factors E2F.

[The intracellular degradation of 125I-EGF and the release of low-molecular degradation products under conditions of actin cytoskeleton disorganization]. / Vnutrikletochnaia degradatsiia 125I-EFR i vyvedenie nizkomolekuliarnykh produktov degradatsii v usloviiakh dezorganizatsii aktinovogo tsitoskeleta.

The influence of cytochalasin B (CB) on the epidermal growth factor (EGF) intracellular degradation and release of low-molecular products of degradation into the culture medium were investigated using two cell lines, A431 and 3T3. Investigated parameters were registered 3.0-4.5 h after the beginning of 125I-EGF endocytosis at 37 degrees C. With A431 cells it was shown that actin cytoskeleton disorganization significantly reduced the rate of 125I-EGF final degradation: a high amount of 125I-EGF, still retained cell-associated, and a reduced amount of low-molecular degradation products were released into the medium in experiments with CB addition. That is in agreement with our previous results (Barkan et al., 1988), revealing a long-term accumulation of EGF-rhodamine fluorescence in a juxtanuclear area of A431 cells after CB treatment. Nevertheless when similar experiments were performed on Swiss 3T3 cells, previously used as a model for demonstrating the inhibitory effect of CB on DNA synthesis stimulation by EGF (Barkan, Nikol'skii, 1986), we could not find such a distinct influence of CB on the process of degradation of internalised 125I-EGF.

[The mitogenic effect of insulin isolated from the mammalian brain on Swiss 3T3 cells]. / Mitogennoe vliianie insulina, vydelennogo iz mozga mlekopitaiushchikh, na kletki Swiss 3T3.

Mitogenic properties of the insulin derived from pig brain were compared with the action of pancreatic (standard) pig insulin and epidermal growth factor (EGF) using the culture of Swiss 3T3 cells. The brain insulin, likely as the pancreatic insulin, induced uptake of 14C-thymidine by resting cells in a dose-dependent manner at concentration of 0.5-2.0 micrograms/ml in culture medium. However, at equal concentrations of these hormones the proliferating effect of the brain insulin appeared to be 2-fold higher than the effect of the pancreatic hormone. At the same time the mitogenic action of both hormones was lower than that of EGF (10 ng/ml). The additive effect of the brain insulin and EGF, administered in combination, was more pronounced than the effect of the pancreatic insulin combined with EGF. The data obtained suggest a possible participation of brain insulin in the process of nerve cell proliferation.

[Certain biochemical characteristics of an insulin-like substance from the bivalve mollusk Anodonta cygnea]. / Nekotorye biokhimicheskie kharakteristiki insulinopodobnogo veshchestvadvustvorchatogo molliuska Anodonta cygnea.

An insulin-like substance (ILS) was isolated from the visceral organs of the bivalve mollusc Anodonta cygnea by chromatography on a sulfocationite CU-23 and purified by reverse phase liquid chromatography. ILS was shown to be made up to several fractions with Mr ranging from 9 to 20 kDa which have identical amino acid composition but different hydrophobicity and N-terminal amino acids. It was supposed that the heterogeneity of ILS fractions is due to its genetical or posttranslational polymorphism. ILS has a low (0.02%) affinity for the mammalian insulin receptor and a low immune affinity for mammalian insulin and possesses a mitogenic activity which is commensurate with that of the epidermal growth factor. The data obtained suggest that Anodonta cygnea ILS represents a separate branch of a relatively ancient family of insulin-like hormones and growth factors responsible for metabolism and proliferation of invertebrate tissues.

[The stimulation of DNA synthesis in resting Swiss 3T3 cells by non-histone chromosomal proteins]. / Stimuliatsiia sinteza DNK v pokoiashchikhsia kletkakh Swiss 3T3 negistonovymi khromosomal'nymi belkami.

Nonhistone chromosomal proteins (NHC), isolated from the kidney and liver of intact rats, the liver of rats treated with hepatocarcinogen DEN and the rat hepatoma, stimulate DNA synthesis in Swiss 3T3 cells in resting culture. The maximum stimulating effect was obtained in the presence of narrow NHC fractions eluted with 0.4-0.5 M NaCl from the phosphocellulose column and identified as hetero-organic NHC protein antigens of the kidney origin associated with hepatocellular tumors.

[Endocytosis and intracellular transport of epidermal growth factor after destruction of the actin cytoskeleton by cytochalasin B]. / Endotsitoz i vnutrikletochnyi transport épidermal'nogo faktora rosta pri destruktsii aktinovogo tsitoskeleta tsitokhalazinom B.

Endocytosis of the epidermal growth factor (EGF) was investigated in three cell lines--A431, 3T6 and Swiss 3T3--after their incubation with cytochalasin B (CB). CB was introduced into culture medium (10 mkg/ml) 1.5-2 hours before addition of 125I-EGF (20-40 ng/ml). The label uptake rate was measured after a 35-40 minutes incubation of cells with 125I-EGF. It appeared that disorganization of microfilamentous network caused by CB exerted no influence on the binding of EGF to the surface membrane receptors and its internalization. Nevertheless, the experiments performed on A431 cells using a fluorescent label--rhodamine--bound to EGF (EGF-R) indicate that CB, though not influencing the initial steps of endocytosis, inhibits the next step--the intracellular transport of EGF-receptor complexes from the trans-Golgi region to lysosomes. As was shown elsewhere (Barkan, Nikol'sky, 1986), CB inhibits the mitogenic effect of EGF on resting Swiss 3T3 cells. So, the process of EGF-receptor uptake and delivery to the trans-Golgi region is evidently not enough to stimulate the cell proliferation; next steps of transport and degradation of ligand-receptor complexes are presumably needed.

[Why does a cell monolayer replicate the metal tray pattern of the thermostat?]. / Pochemu kletochnyi monosloi vosproizvodit risunok metallicheskoi polki termostata?

Cells freshly seeded into the closed culture flasks or dishes and placed on the metal tray with holes of the thermostat or incubator are seen to form the layer with uneven density: with high density corresponding to the flask bottom regions above the metal and low density corresponding to the flask bottom region above the holes in the tray. The effect was shown using several cell lines with different degrees of transformation and saturation density, including Swiss 3T3. The main cause of this effect is the difference between the temperature inside the thermostat and the lower temperature of the flasks with culture medium (rather than between the metal framework and the air), together with a high heat conduction of the metal. The reverse difference in the temperatures (higher temperature of the culture flasks) leads to the formation of the reverse pattern of the cell layer, with higher density corresponding to the holes. The temperature differences exert their influence presumably during the first 10-15 minutes after the cells seeding, when the process of cell sedimentation is involved possibly by creating the microcurrents in the medium.

[Effect of dihydrocytochalasin B on the induction of DNA synthesis by epidermal growth factor, insulin and serum in Swiss 3T3 cells]. / Vliianie digidrotsitokhalazina B na induktsiiu sinteza DNK épidermal'nym faktorom rosta, insulinom i syvorotkoi v kletkakh Swiss 3T3.

The addition of a microfilament-disorganizing agent--dihydrocytochalasin B B (5-10 micrograms/ml)--to to quiescent confluent or sparse (in 0.5% serum) Swiss 3T3 cells, 1-2 hours prior to stimulation, inhibited the initiation of DNA synthesis induced by an epidermal growth factor (7.5-10 ng/ml) and insulin (0.5-1.0 micrograms/ml), but exerted a low effect on serum stimulation. DNA synthesis was measured 21-23 hours after the growth factor administration by 14C-thymidine incorporation in acid-insoluble material and the ratio of this fraction to exogenous thymidine uptake. Moreover, the polar solvent dimethylsulfoxide, present in culture medium at low concentration (0.1-0.5%), also caused a decrease in the basal level of 14C-thymidine incorporation in resting cells, and a less decrease in the induced incorporation.

[Amine inhibition of serum-induced DNA synthesis in 3T3 and 3T6 cell cultures]. / Podavlenie aminami sinteza DNK, indutsirovannogo syvorotkoi v kul'ture kletok 3T3 i 3T6.

Several amines were shown to inhibit growth stimulation in quiescent confluent and sparse cultures of Swiss 3T3 and 3T6 cells by changing for a fresh medium containing 10-20% serum. Proliferation was inhibited by dansylcadaverine (0.1 mM), chloroquine (0.05 mM), 5-methoxytryptamine (0.1 mM), cystamine (0.1 mM), dimethylurea (100 mM), methylurea (100 mM), and in some experiments--by urea (100 mM). The inhibitory activity was not associated with a direct influence of amines on DNA synthesis or thymidine phosphorylation. The findings suggest that amines may influence the process of clustering of growth factor-receptor complexes, or the fusion of plasma membrane and intracellular vesicles containing some components required for cell proliferation.

[Minimally transformed 3T3 cell lines as an object for research on the mechanisms of proliferation]. / Minimal'no transformirovannye kletochnye linii 3T3 kak ob"ekt issledovaniia mekhanizmov proliferatsii.

The present review gives a detailed description of mouse cell lines 3T3 (Swiss 3T3, Balb/c 3T3, and NIH 3T3) including their establishment, evolution, growth properties and formation of the specific monolayer with low saturation density. Questionable views of the 3T3 cell origin are discussed. The main features of these continuous cell lines are shown, such as a high requirement for serum growth factors, substrate dependence, inhibition of cell division in dense monolayer and absence of oncogenic potentions, which are similar to those of normal diploid cells, thus making these pseudonormal lines useful in studies of cell proliferation and oncogenesis. Different methods for obtaining quiescent 3T3 cells are observed, and, more in detail, the achievement of the resting state in confluent monolayer and at low serum concentration in sparce culture. The data reporting the efficiency of growth stimulation using serum, purified growth factors and hormones are analysed. Special attention is paid to the susceptibility of 3T3 lines to culture conditions, and to dependence of experimental data obtained upon keeping stability of these conditions.

[Effect of cultivation conditions on the growth properties of Swiss 3T3 cells]. / Vliianie uslovii kul'tivirovaniia na rostovye svoistva kletok Swiss 3T3.

A possibility of using Swiss 3T3 cells, adapted to the growth in the Eagle basal medium and bovine serum, in studies of cell proliferation and quiescent state was shown on the basis of their growth characteristics. Proliferative activity of cultures was estimated by measuring the intensity of DNA synthesis (incorporation of labeled thymidine and flow cytofluorometric analysis), mitotic index and cell number counts. Growth rate and saturation density of the culture were analyzed depending on serum concentration, substrate quality and medium changes both in growth and quiescent states. In spite of repeated medium changes such adapted cells had saturation density within 4.10(4)--7.10(4) cells/cm2, standard for this line. Besides, a distinct inhibition of cell proliferation at confluence or after incubation with low serum (0.5%) and a possibility of the following stimulation of cell divisions by adding a fresh medium containing different concentrations of serum were demonstrated. The increased rate of adipose conversion was detected in resting confluent 3T3 cells cultivated in closed vessels, as compared to cells growing in tissue culture dishes in the CO2 incubator.