Inhibition of gap junction and adherens junction assembly by connexin and A-CAM antibodies

We examined the roles of the extracellular domains of a gap junction protein and a cell adhesion molecule in gap junction and adherens junction formation by altering cell interactions with antibody Fab fragments. Using immunoblotting and immunocytochemistry we demonstrated that Novikoff cells contained the gap junction protein, connexin43 (Cx43), and the cell adhesion molecule, A-CAM (N-cadherin). Cells were dissociated in EDTA, allowed to recover, and reaggregated for 60 min in media containing Fab fragments prepared from a number of antibodies. We observed no cell-cell dye transfer 4 min after microinjection in 90% of the cell pairs treated with Fab fragments of antibodies for the first or second extracellular domain of Cx43, the second extracellular domain of connexin32 (Cx32) or A-CAM. Cell-cell dye transfer was detected within 30 s in cell pairs treated with control Fab fragments (pre-immune serum, antibodies to the rat major histocompatibility complex or the amino or carboxyl termii of Cx43). We observed no gap junctions by freeze-fracture EM and no adherens junctions by thin section EM between cells treated with the Fab fragments that blocked cell-cell dye transfer. Gap junctions were found on approximately 50% of the cells in control samples using freeze-fracture EM. We demonstrated with reaggregated Novikoff cells that: (a) functional interactions of the extracellular domains of the connexins were necessary for the formation of gap junction channels; (b) cell interactions mediated by A-CAM were required for gap junction assembly; and (c) Fab fragments of antibodies for A-CAM or connexin extracellular domains blocked adherens junction formation

Antioxidants for Industrial Applications

The thermooxidative degradation of organic materials as well as its inhibition by classical and nonclassical antioxidants are briefly outlined

INDUCTION OF APOPTOSIS BY FATTY ACID RICH FRACTION OF SOLANUM NIGRUM ON CERVICAL CANCER CELL LINES

Objective: Mechanism of cell death inducing pathway of chloroform fraction of methanolic extract of Solanum nigrum were studied in a panel of cervical cancer cell line.Methods: Cells were treated with IC50 doses (previously determined by MTT assay), and various experiments were performed. For cellular and nuclear morphology, cells were stained with Hoechst33258 and observed under phase and fluorescence microscopy. Cell cycle shift and apoptotic assay (AnnexinV-FITC/PI) were studied by flow cytometry. Expressions of relevant genes were studied at the transcriptional and translational levels. ROS induction and loss of MMP and presence of γH2AX were studied by fluorescent microscopy. Constituents of the fraction were separated by column chromatography and bioassayed by MTT assay. Bioactive sub-fractions were identified by GC-MS.Results: Typical apoptotic morphological features were found in the treated cells. Most cells were found in late apoptotic stages. In HeLa and C33A cell lines, cells were blocked at G1/S, in SiHa, subG0 population increased. Expression of antiapoptotic Bcl-2 decreased and that of proapoptotic BAX, p53, p21 increased. Expression of HPV16 and human ET1 also decreased. ROS mediated DNA damage was detected and induced apoptosis. Different fatty acids along with other compounds were detected in the chloroform fraction, all of which are reported to have anticancer properties.Conclusion: From the study it can be concluded that fatty acid rich chloroform fraction of S. nigrum is capable of inducing apoptosis in cervical cancer cells, through ROS mediated DNA damage

Narrative Skills in 4-Year-Olds With Normal, Impaired, and Late-Developing Language

Two groups of children who were slow in expressive language development (SELD) at age 2 and a matched group of toddlers with normal language were re-evaluated at age 4. Assessment included measures of productive syntactic skills in spontaneous speech and narrative abilities in a standard story retelling task. Four-year-olds who continued to perform below the normal range in sentence structure production scored significantly lower than their normally speaking peers on all measures of narrative skill. Children who were slow to begin talking at age 2 but who, by age 4, had moved into the normal range in basic sentence structure production showed no statistically significant differences, in terms of several of the measures of narrative ability, from either normally speaking 4-year-olds or from the group with persistent delay. The implications of these findings for the management of early language delay and its relation to school learning disability are discussed

Evaluation of Semi-Automate Fluoro-Kit for Fta-Abs Testing

A semi-automated Fluoro-Kit for FTA-Abs testing was evaluated in a double-blind study Identical Automated Reagin Test (ART) and Venereal Disease Research Lab (VDRL) positive specimens obtained during an 11-month period of routine admission serologic tests for syphilis were tested at Saint Barnabas Medical Center by the Clinical Sciences Semi-Automated Fluoro-Kit for FTA-Abs testing. Aliquots of these specimens were also sent to a reference laboratory for confirmation. The results of the study showed that the Clinical Sciences Fluoro-Kit was reliable test system for the confirmatory FTA-Abs testing of both ART and VDRL, reactive sera. In addition, the semi-automated Fluoro-Kit offers a significant advance in standardization and in the relative speed and economy of performance of FTA-Abs tests

What is the best treatment for plant-induced contact dermatitis?

It's unclear which treatment is best, because there have been no head-to-head comparisons of treatments for Rhus (plant-induced) contact dermatitis. That said, topical high-potency steroids slightly improve pruritus and the appearance of the rash (strength of recommendation [SOR]: B, small cohort studies). Neither topical pimecrolimus (an immunomodulatory drug) nor jewelweed extract are helpful (SOR: B, 1 small randomized controlled trial [RCT]). Oral steroids improve symptoms in severe cases (SOR: C, expert opinion)

INDUCTION OF AUTOPHAGY IN HUMAN CERVICAL CANCER CELL LINE (SIHA) BY CHAETOMORPHA LINUM (MULLER) KÃœTZ.

Objective: Algae is a rich source of various metabolites. Use of algae in traditional medicine was reported from very early days. The potential anticancerous activity of the algae was also reported by recent researches. In the present study, cell death mechanism induced by green alga Chaetomorpha linum (Muller) Kütz. was studied on human cervical cancer cell line, SiHa (Human papilloma virus-HPV 16+ve).Methods: Cells were treated with chloroform fraction of the Chaetomorpha linum (CLC). DNA fragmentation, cell proliferation, nuclear morphology, localization of autophagosomes and expression of relevant proteins were studied. Cell cycle and mitochondrial membrane potential (MMP) were assessed by fluorescence assisted cell sorting (FACS). Gas chromatography liquid spectrometry (GCMS) analysis of CLC was done to detect the algal compounds.Results: Preliminary experiments confirmed the absence of DNA fragmentation and altered nuclear morphology in the treated cells. In the Acridine Orange stained cells, the presence of autophagosomes was observed by both microscopy and FACS analysis. Decreased MMP was observed in the treated sets. Wound healing assay showed the inhibitory activity of CLC on SiHa cells. Increased expression of autophagic proteins, viz; LC3BII, p62/SQSTM, Beclin-1 and decreased expression of S6 were observed in the treated sets; indicating that autophagic cell death was induced. Preliminary qualitative chemical study and GCMS analysis detected the presence of saponins, glycosides, alkaloids, phenols, steroids and fatty acids in CLC.Conclusion: All these findings clearly suggested that the chloroform fraction of the algae was responsible for induction of apoptosis followed by autophagic cell death in SiHa and can serve as a potential candidate for anticancer drug development in future

Key lesion localization and pre-surgical planning using magnetoencephalography in patients with medically refractory epilepsy: three case reports.

Epilepsy is a common and debilitating disease affecting up to 50 million people worldwide. Nearly 30% of patients with epilepsy have disease refractory to treatment with medication alone. Even in medically refractory disease, neurosurgical resection can be curative when the epileptic focus is correctly identified. Several non-invasive techniques are available for epileptic focus localization and pre-surgical planning. These include electroencephalography (EEG), magnetic resonance imaging (MRI), and magnetoencephalography (MEG).Each of these techniques provides complementary information for precise lesion localization and targeted neurosurgical approach to minimize damage to important adjacent structures. We present three cases of medically refractory epilepsy. The cases show how the combination of EEG, MRI, and MEG allows for lesion localization and safe surgical planning in a variety of cases. They include epilepsy related to cortical dysplasia, grey matter heterotopia, and tumor recurrence. We emphasize the role of MEG and demonstrate how it can provide critical additional information which is not captured by conventional EEG and MRI alone

Fears of compassion in a depressed population: Implication for psychotherapy

Background: While psychological therapies for depression have advanced in the last 20 years, still many people respond only partially and remain vulnerable to relapse. Insight into the limitations of our psychological therapies might be obtained from recent research that has revealed, in nonclinical populations, that some people can be fearful of positive emotions especially affiliative and compassion-focused ones. Aims: This study explores the fears of compassion in a clinical population and their associations with selfcriticism, self-compassion and depression, anxiety and stress. Method: 53 depressed patients completed a series of self-report scales. Results: Fears of compassion, particularly for oneself and from others, were strongly linked to self-criticism, depression, anxiety and stress, and negatively associated with self-compassion and self-reassurance. Conclusions: Since compassion and the affiliative emotions associated with compassion play a fundamental role in emotion regulation, individuals who are blocked or fearful of accessing these emotions are likely to be struggle with emotional regulation and the psychotherapeutic process. Research on the fears of compassion and affiliative emotions suggests these are important therapeutic targets.N/