Kaiso depletion attenuates transforming growth factor-ß signaling and metastatic activity of triple-negative breast cancer cells.

Triple-negative breast cancers (TNBCs) represent a subset of breast tumors that are highly aggressive and metastatic, and are responsible for a disproportionate number of breast cancer-related deaths. Several studies have postulated a role for the epithelial-to-mesenchymal transition (EMT) program in the increased aggressiveness and metastatic propensity of TNBCs. Although EMT is essential for early vertebrate development and wound healing, it is frequently co-opted by cancer cells during tumorigenesis. One prominent signaling pathway involved in EMT is the transforming growth factor-ß (TGFß) pathway. In this study, we report that the novel POZ-ZF transcription factor Kaiso is highly expressed in TNBCs and correlates with a shorter metastasis-free survival. Notably, Kaiso expression is induced by the TGFß pathway and silencing Kaiso expression in the highly invasive breast cancer cell lines, MDA-MB-231 (hereafter MDA-231) and Hs578T, attenuated the expression of several EMT-associated proteins (Vimentin, Slug and ZEB1), abrogated TGFß signaling and TGFß-dependent EMT. Moreover, Kaiso depletion attenuated the metastasis of TNBC cells (MDA-231 and Hs578T) in a mouse model. Although high Kaiso and high TGFßR1 expression is associated with poor overall survival in breast cancer patients, overexpression of a kinase-active TGFßR1 in the Kaiso-depleted cells was insufficient to restore the metastatic potential of these cells, suggesting that Kaiso is a key downstream component of TGFß-mediated pro-metastatic responses. Collectively, these findings suggest a critical role for Kaiso in TGFß signaling and the metastasis of TNBCs.

The reliability and accuracy of an electromagnetic motion analysis system when used conjointly with an accelerometer.

The effect of an accelerometer driven electronic postural monitor (Spineangel®) placed within the electromagnetic measurement field of the Polhemus Fastrak™ is unknown. This study assessed the reliability and accuracy of Fastrak™ linear and angular measurements, when the Spineangel® was placed close to the sensor(s) and transmitter. Bland Altman plots and intraclass correlation coefficient (2,1) were used to determine protocol reproducibility and measurement consistency. Excellent reliability was found for linear and angular measurements (0.96, 95% CI: 0.90-0.99; and 1.00, 95% CI: 1.00-1.00, respectively) with the inclusion of Spineangel®; similar results were found, without the inclusion of Spineangel®, for linear and angular measurements, (0.96, 95% CI: 0.89-0.99; and 1.00, 95% CI: 1.00-1.00, respectively). The greatest linear discrepancies between the two test conditions were found to be less than 3.5 mm, while the greatest angular discrepancies were below 3.5°. As the effect on accuracy was minimal, these findings support the conjoint use of the Fastrak™ during validation studies of the Spineangel® device. STATEMENT OF RELEVANCE: Although previous studies have used the Fastrak™ as the gold standard measurement system, the influence of an accelerometer driven postural monitor on accuracy has not been reported. The strength of the present study has been to determine the effect of accelerometer placement within the electromagnetic field on the reliability and accuracy of the Fastrak™.

Ethnobotanical study on traditional uses of wild medicinal plants in Prokletije Mountains (Montenegro).

AIM OF THE STUDY: The main objectives were to collect information on the use of wild growing medicinal plants by local people living in high mountain region of Montenegro and conduct local botanical and ecological surveys. Active ingredients of plant species officinal in European Pharmacopoeia 6.0 (Ph. Eur. 6.0) were studied and we assessed possibilities for commercial exploitation for local economic development. MATERIALS AND METHODS: The 75 people that were interviewed (40-82 years old) identified 94 species for treatment of various human ailments. For each named species, the following elements are provided: botanical name, family, part(s) used, medicinal use and perceived property, listing in published pharmacopoeias, the relative abundance of each species and locality where the plant was collected. Chemical analyses were done according to prescriptions of Ph. Eur. 6.0 in order to estimate potential commercial use of native plants. RESULTS AND CONCLUSION: The most common in traditional usage were Rosaceae (11 species) making 11.7%, Asteraceae (10 species) 10.6% and Lamiaceae (7 species) 7.4%. From 94 species reported, 35 (37.2%) are officinal in Ph. Eur. 6.0 and 12 in national pharmacopoeias (12.8%). Aerial parts were mostly used (43.6%). The most frequently reported medicinal uses were for treating gastrointestinal (57.4%) and respiratory diseases (41.5%).

The relationship between physical activity and low back pain outcomes: a systematic review of observational studies.

Although clinical guidelines advocate exercise and activity in the management of non-specific low back pain (NSLBP), the link between levels of physical activity and outcomes is unclear. This systematic review investigated the relationships between free living activity levels after onset of low back pain (LBP) and measures of pain, and disability in patients with NSLBP. Cohort and cross-sectional studies were located using OVID, CINAHL, Medline, AMED, Embase, Biomed, PubMed-National Library of Medicine, Proquest and Cochrane Databases, and hand searches of reference lists. Studies were included if a statistical relationship was investigated between measures of free living physical activity (PA) in subjects with LBP and LBP outcome measures. Twelve studies (seven cohort and five cross-sectional) were included. One prospective study reported a statistically significant relationship between increased leisure time activity and improved LBP outcomes, and one cross-sectional study found that lower levels of sporting activity were associated with higher levels of pain and disability. All other studies (n = 10) found no relationship between measures of activity levels and either pain or disability. Heterogeneity of study designs, particularly in terms of activity measurement, made comparisons between studies difficult. These data suggest that the activity levels of patients with NSLBP are neither associated with, nor predictive of, disability or pain levels. Validated activity measurement in prospective research is required to better evaluate the relationships between PA and LBP.

Can postural modification reduce kinetic and kinematic loading during the bowing postures of Islamic prayer?

As stooped postures are known to increase kinematic and kinetic loading on the lumbar spine they can be problematic for people with low back pain and postural task modification is often recommended. For the Muslim with low back pain, the bowing postures during prayer can aggravate low back symptoms. The aims of this study were to describe lumbo-sacral and pelvic tilt kinematics and lumbo-sacral kinetics during the standard bowing postures of Islam and to compare these to kinematic and kinetic data gathered during a clinically recommended modified bowing posture. The study was a repeated measures within subject cross-over design with 33 healthy male Muslim participants. 3-D motion analysis data were gathered to calculate body joint angles during the two bowing postures. A 3-D biomechanical model was then used to calculate spinal loads. Paired t-test analyses showed that the use of the modified posture resulted in significantly less pelvic tilt range of motion and anterior shear force and compressive force L5/S1, at stages 1 and 5 of bowing. Although this study was conducted with healthy young Muslim males, the use of this modified bent knee posture is recommended for all Muslims with low back pain. Clinical trials are being considered to determine the clinical utility of this postural manoeuvre as an intervention. STATEMENT OF RELEVANCE: The presence of low back pain may hinder a Muslim's ability to use the traditional Islamic bowing posture. Muslims who have low back pain may benefit from adopting a modification to the traditional bowing posture, which has been found to reduce the loads and postural demands on the lower back.

The effectiveness of walking as an intervention for low back pain: a systematic review.

As current low back pain (LBP) guidelines do not specifically advocate walking as an intervention, this review has explored for the effectiveness of walking in managing acute and chronic LBP. CINAHL, Medline, AMED, EMBASE, PubMed, Cochrane and Scopus databases, as well as a hand search of reference lists of retrieved articles, were searched. The search was restricted to studies in the English language. Studies were included when walking was identified as an intervention. Four studies met inclusion criteria, and were assessed with a quality checklist. Three lower ranked studies reported a reduction in LBP from a walking intervention, while the highest ranked study observed no effect. Heterogeneity of study design made it difficult to draw comparisons between studies. There is only low-moderate evidence for walking as an effective intervention strategy for LBP. Further investigation is required to investigate the strength of effect for walking as a primary intervention in the management of acute and chronic LBP.

Can accelerometry be used to discriminate levels of activity?

The aims of this study were to investigate the associations between an activity logbook and the RT3 accelerometer and to assess whether the RT3 can discriminate activity levels in healthy adults. Ten participants completed two trials wearing an RT3 accelerometer over a 4-6 h period and completed a detailed activity log. Results showed a poor correlation between the RT3 in moderate activities (r = 0.22) in comparison to low (r = 0.52) and hard (r = 0.70) from the logbook. A significant difference was found in average RT3 vector magnitude (VM) counts/min in each activity level (p < 0.0001). Discriminant analysis demonstrated that an RT3VM counts/min value of approximately 500 was found to have high sensitivity (88%), and specificity (88%) for discriminating between low and moderate activity levels from the logbook. This study found that accelerometry has the potential to discriminate activity levels in free living. This study is the first to investigate whether tri-axial accelerometry can discriminate different levels of free-living activity recorded in an activity logbook. The RT3 accelerometer can discriminate between low and moderate physical activities and offers a methodology that may be applicable to future research in occupational settings.

Some wild-growing plant species from Serbia and Montenegro as the potential sources of drugs.

The results of phytochemical investigation, over the last decade, of some wild-growing plant species from Serbia and Montenegro belonging to the families Asteraceae, Guttiferae and Gentianaceae are reported. Most of the investigated species are endemites and the emphasis in this report is on those exhibiting biological activities that could be regarded as the potential sources of drugs. This review comprises 154 compounds, e.g. sesquiterpene lactones and flavonoids (Asteraceae), xanthones, secoiridoids and C-glucoflavonoids (Gentanaceae) and prenylated phloroglucinols (Guttiferae) as well as some other secondary metabolites, produced by the above families, which could be of pharmacological interest.

Blockade of central vasopressin receptors reduces the cardiovascular response to acute stress in freely moving rats.

To investigate the contribution of central vasopressin receptors to blood pressure (BP) and heart rate (HR) response to stress we injected non-peptide selective V(1a) (SR49059), V(1b) (SSR149415), V(2) (SR121463) receptor antagonists, diazepam or vehicle in the lateral cerebral ventricle of conscious freely moving rats stressed by blowing air on their heads for 2 min. Cardiovascular effects of stress were evaluated by analyzing maximum increase of BP and HR (MAX), latency of maximum response (LAT), integral under BP and HR curve (integral), duration of their recovery and spectral parameters of BP and HR indicative of increased sympathetic outflow (LF(BP) and LF/HF(HR)). Moreover, the increase of serum corticosterone was measured. Exposure to air-jet stress induced simultaneous increase in BP and HR followed by gradual decline during recovery while LF(BP) oscillation remained increased as well as serum corticosterone level. Rats pre-treated with vasopressin receptor antagonists were not sedated while diazepam induced sedation that persisted during exposure to stress. V(1a), V(1b) and V(2) receptor antagonists applied separately did not modify basal values of cardiovascular parameters but prevented the increase in integral(BP). In addition, V(1b) and V(2) receptor antagonists reduced BP(MAX) whereas V(1a), V(1b) antagonist and diazepam reduced HR(MAX) induced by exposure to air-jet stress. All drugs shortened the recovery period, prevented the increase of LF(BP) without affecting the increase in serum corticosterone levels. Results indicate that vasopressin receptors located within the central nervous system mediate, in part, the cardiovascular response to air-jet stress without affecting either the neuroendocrine component or inducing sedation. They support the view that the V(1b) receptor antagonist may be of potential therapeutic value in reducing arterial pressure induced by stress-related disorders.

Dynamic changes in CD4+ CD25+(high) T cell apoptosis after the diagnosis of type 1 diabetes.

Because type 1 diabetes (T1D) is a chronic, autoimmune, T cell-mediated disease, interventions affecting T cells are expected to modulate the immune cascade and lead to disease remission. We propose that increased CD4(+) CD25(+high) T cell apoptosis, a trait we discovered in recent-onset T1D subjects, reflects T1D partial remission within the first 6 months after diagnosis. Apoptosis of forkhead box P3 (FoxP3)(+) CD4(+) CD25(+high) T cells, in addition to total daily doses of insulin (TDD), blood glucose, HbA1c and age, were measured in 45 subjects with T1D at various times after diagnosis. Sixteen healthy control subjects were also recruited to the study. Higher CD4(+) CD25(+high) T cell apoptosis levels were detected within the first 6 months of diagnosis (odds ratio = 1.39, P = 0.009), after adjustment for age, TDD and HbA1c. A proportional hazards model confirmed that the decline of apoptosis after diagnosis of T1D was related significantly to survival time (hazards ratio = 1.08, P = 0.014), with TDD and age also contributing to survival. During this time there was an inverse relationship between CD4(+) CD25(+high) T cell apoptosis with TDD (r = -0.39, P = 0.008). The CD4(+) CD25(+high) T cell apoptosis levels decline significantly after the first 6 months from diagnosis of T1D and may help in the close monitoring of autoimmunity. In parallel, there is an increase in TDD during this time. We also propose that CD4(+) CD25(+high) T cell apoptosis assay can be used to gauge the efficacy of the several immune tolerance induction protocols, now under way.

Complete assignments of 1H and 13C NMR spectra of leucanthoside A, a new triterpenoid saponin from Cephalaria leucantha L.

Leucanthoside A, a new allose-containing triterpenoid saponin (1), was isolated from the aerial parts of Cephalaria leucantha L. Its structure was determined by electrospray ionization mass spectrometry and NMR spectroscopy. Complete assignments of the 1H and 13C NMR chemical shifts were achieved by two-dimensional NMR experiments: DQF-COSY, NOESY, TOCSY, HSQC, DINE-HSQC, HMBC, 13C-1H 2D-J-resolved spectroscopy, and 1,1-ADEQUATE.

Comparison of apoptosis and mortality measurements in peripheral blood mononuclear cells (PBMCs) using multiple methods.

Death through apoptosis is the main process by which aged cells that have lost their function are eliminated. Apoptotic cells are usually detected microscopically by changes in their morphology. However, determination of early apoptotic events is important for in vitro (and ex vivo) studies. The main objective of the present study is to find the most sensitive method for apoptosis detection in human peripheral blood mononuclear cells (PBMCs) by comparing six different methods following five different means of immunological stimulation at 3 and 5 days. Each of six apoptosis quantification methods, except the trypan blue exclusion test, is a combination of two stains, one for the specific detection of apoptotic cells and the other for the unspecific detection of dead cells. Values for apoptosis and mortality were compared with a reference method. The choice of apoptosis detection method is more important following 3 days of stimulation than after 5 days of stimulation (P=2x10(-6) versus P=1x10(-2)). In contrast, we find mortality measurements following the different means of stimulation highly significant at both 3 and 5 days (F2.28=7.9, P=1.4x10(-6) at 3 days and F2.28=8.5, P=4.5x10(-7) at 5 days). Variation as a result of the combination of specific PBMC stimulation and the method used to detect apoptosis is reduced considerably with time (F1.58+3.7, P+3x10(-7) at 3 days to F=1.58=0.97, P=0.5 at 5 days). Based on Tukey's test, YO-PRO-1 is the most sensitive stain for apoptosis and, when combined with 7-AAD, provides an accurate measure of apoptosis and mortality. In conclusion, we propose YO-PRO-1/7-AAD as a new combination and low-cost alternative for the sensitive detection of early apoptosis.

Further degradation product of hyperforin from Hypericum perforatum (St John's Wort).

Repeated examination of the aerial parts of Hypericum perforatum yielded a new degradation product of hyperforin (1) namely deoxyfurohyperforin A (2), together with the previously identified furohyperforin (3), furoadhyperforin (4), furohyperforin A (5a and 5b), pyrano[7,28-b]hyperforin (6) and 3-methyl-4,6-di(3-methyl-2-butenyl)-2-(2-methyl-1-oxopropyl)-3-(4-methyl-3-pentenyl)-cyclohexanone (7). The antimicrobial activity of the compounds 3, 5a and 5b, 6 and 7 was tested against Staphylococcus aureus, Candida albicans, Bacillus subtilis and Escherichia coli.

Sesquiterpene lactones from the aerial parts of Inula oculus-christi.

Pulchellin E (1) and gaillardin (2) were isolated from the aerial parts of Inula oculus-christi, along with the flavone hispidulin. The 13C-NMR chemical shifts of 1 and 2 are reported.

Mixed cobalt(III) complexes with aromatic amino acids and diamine. Part IV. Influence of noncovalent intra-and interligand interactions on rotamer population of the S-phenylalaninato and S-tyrosinato side groups in (1,2-diaminoethane)bis-(S-aminocarboxylato)cobalt(III) complexes.

Rotamer population of S-tyrosinato and S-phenylalaninato ligands side groups in diastereomers of (1,2-diaminoethane)bis-(S-aminocarboxylato)cobalt(III) complexes is calculated by vicinal alpha and beta proton coupling constant analysis. The effect of noncovalent intra- and interligand interactions on the population of rotamers in D20 solution is discussed. It has been established that in all the complexes investigated the most abundant is rotamer t, in which aromatic voluminous moiety and carboxylic group are in an anti position. In almost all complexes the lowest content is of rotamer g, in which these two groups are in the nearest position. Relatively high population of rotamer h in complex 5 tyr, in spite of high steric hindrances, is due to intra- and interligand NH...pi interactions.

Variability of essential oils of Satureja montana L. and Satureja kitaibelii wierzb. ex Heuff. from the central part of the balkan peninsula.

The results of the essential oil composition confirm that there are differences between populations within the same subspecies (Satureja montana ssp. montana, between the subspecies (Satureja montana ssp. montana and Satureja montana ssp. pisidica) and the species (Satureja montana and S. kitabelii). The essential oil composition of S. montana ssp. montana is more related to the Satureja montana ssp. pisidica than S. kitaibelii which separates as a whole species.

Mixed cobalt(III) complexes with aromatic amino acids and diamine. Part II. Synthesis and configuration of (1,2-diaminoethane)bis(L-tyrosinato)cobalt(III)complex diastereomers. The crystal structure of the delta-C1-cis(O) diastereomer.

In the reaction of trans-[CoCl2(en)2]+ with L-tyrosine all six theoretically possible diastereomers of the (1,2-diaminoethane)bis(L-tyrosinato)cobalt(III) complex were formed. The following five were isolated: gamma-trans(O); and gamma- and delta-C2-cis(O) and gamma- and delta-C1-cis(O) diastereomers, while the delta-trans(O) diastereomer was only detected in the corresponding eluate. Separation of the obtained diastereomers was performed by chromatography on a Dowex 1 x 4 column. Characterization of the isolated diastereomers was carried out by means of elemental analysis, electronic absorption, circular dichroic, 1H and 13C NMR spectra, and by x-ray crystal structure analysis in the case of the delta-C1-cis(O) diastereomer. We established the general rule of preference of diasteromers formation in complexes of [Co(L-aa)2diamine]+ (L-aa = L-amino acid anion; diamine = 1,2-diaminoethane or 1,3-diaminopropane) type.

Further sesquiterpene lactones from Anthemis carpatica.

A new germacranolide, (E)-1alpha, 10beta-epoxy-3beta-acetoxy-6alpha-hydroxygermacra-4,11 (13)-dien-12,8alpha-olide, together with nine new highly oxygenated guaiadien-12,6alpha-olides of anthemolide, and cumambrin type were identified in the repeated examination of the aerial parts of the flowering Anthemis carpatica. In addition, six known guaianolides belonging to the same groups, also isolated previously from A. carpatica, along with two guaianolides, 2beta-hydroxyepiligustrin and cumambrin B, not found before in this species, were isolated this time.