Differentiation of normal and cancer cells induced by sulfhydryl reduction: biochemical and molecular mechanisms.

We examined the morphological, biochemical and molecular outcome of a nonspecific sulfhydryl reduction in cells, obtained by supplementation of N-acetyl-L-cysteine (NAC) in a 0.1-10 mM concentration range. In human normal primary keratinocytes and in colon and ovary carcinoma cells we obtained evidences for: (i) a dose-dependent inhibition of proliferation without toxicity or apoptosis; (ii) a transition from a proliferative mesenchymal morphology to cell-specific differentiated structures; (iii) a noticeable increase in cell-cell and cell-substratum junctions; (iv) a relocation of the oncogenic beta-catenin at the cell-cell junctions; (v) inhibition of microtubules aggregation; (vi) upregulation of differentiation-related genes including p53, heat shock protein 27 gene, N-myc downstream-regulated gene 1, E-cadherin, and downregulation of cyclooxygenase-2; (vii) inhibition of c-Src tyrosine kinase. In conclusion, a thiol reduction devoid of toxicity as that operated by NAC apparently leads to terminal differentiation of normal and cancer cells through a pleiade of converging mechanisms, many of which are targets of the recently developed differentiation therapy.

Production and characterization of antibodies for the specific determination of the opioid peptide Met5-Enkephalin-Arg6-Phe7.

Endogenous opioids serve as modulators of neuroendocrine and immune system processes, the investigation of which calls for high-specificity radioimmunoassays (RIAs). This study focuses on the development and use of a specific radioimmunoassay for the opioid peptide Met5-Enkephalin-Arg6-Phe7 (MEAP), the C-terminus part of proenkephalin A. Antibodies were raised in four rabbits and investigated in terms of titre, avidity and specificity, followed by finding ideal conditions for these antibodies in RIA. MEAP concentrations were determined in crude extracts of rat hypothalamus, dorsal root ganglia, adrenals and ankle using this standardized assay after an oxidizing process. At reverse-phase high-pressure liquid chromatography (HPLC), the position of immunoreactive material from rat hypothalamus eluted as two peaks out of which one was compatible with that of synthetic MEAP. All rabbits exhibited individual differences in relative immune response and time of its onset. The avidity constant was 10 times higher on a molar basis for ab 4108 compared with ab 4182. There was no cross-reactivity for ab 4182 to related peptides, such as enkephalins and dynorphin B, and negligible background values for ab 4108. The relative levels ofimmunoreactive MEAP from the CNS versus peripheral tissues contrasted in accordance with current knowledge. It is suggested that reports with RIA results should include characterization of antibodies, extraction procedures, standard curves and compositions of buffers. Furthermore, the results should preferably be expressed in relation to total protein content.

Effects of acupuncture on skin temperature in children with neurological disorders and cold feet: an exploratory study.

In a single subject design, acupuncture effects on peripheral skin temperature were studied in six children with cold feet associated with cerebral palsy, progressive encephalopathy or Smith-Lemli-Opitz syndrome. Skin temperature was measured at fixed points on the forehead, suprasternal fossa, hand dorsum and palm, foot dorsum and sole from a distance of 3 cm with a Raytek Raynger ST2L. Acupuncture needles were inserted in arms or legs with or without electrical stimulation. Three children displayed a brief rise in temperature in the hands and one in the feet. A tendency towards a cumulative effect in improved skin temperature occurred in two of three children receiving additional sessions. One child did not respond to acupuncture. It is possible that acupuncture may increase skin temperature in some children with cold extremities of neurological origin. Studies in larger groups are needed.

Reliability and responsiveness of three different pain assessments.

The visual analogue scale (VAS) and ordered categorical scales, i.e. numeric rating scales (NRS), are commonly used in the assessment of pain. However, these scales are bounded by fixed endpoints and thus the range of measurement is limited. The disparity in repeated assessments of perceived pain intensity with the VAS, NRS, and electrical stimulation applied as a matching stimulus was studied in 69 patients (48 women and 21 men, 19-72 years) with chronic nociceptive or neurogenic pain. Responsiveness with transcutaneous electrical nerve stimulation (TENS) using the same measurement procedures was evaluated in the same patients. Comparison of results from the three pain assessments showed that the painmatcher is at least as reliable and responsive as VAS and NRS. None of the three measurements showed evidence for systematic disagreement and had only significant random individual disagreement. They also showed evidence for responsiveness.

Effects of electro-acupuncture on corticotropin-releasing factor in rats with experimentally-induced polycystic ovaries.

The aims of the present study were to investigate corticotropin-releasing factor (CRF) concentrations in the brain, the adrenal glands, and the ovaries in rats with estradiol valerate (EV) induced polycystic ovaries (PCO). The effect of 12 electro-acupuncture (EA) treatments on CRF concentrations was also investigated. The CRF concentrations in the median eminence (ME) were significantly increased in rats with PCO (both the PCO control group and the PCO group receiving EA) compared with the healthy control group (veichle control group), indicating increased activity in the hypothalamus-pituitary axis. The CRF concentrations in the ovaries were significantly reduced in the PCO group receiving EA compared with the PCO control group. Also, there was a decrease in comparison withthe healthy control group but the decrease was not as significant. This finding indicates that repeated EA treatments change the neuroendocrinological state in the ovaries, which may play an important role in reproductive failure.

Effects of electro-acupuncture on nerve growth factor and ovarian morphology in rats with experimentally induced polycystic ovaries.

Despite extensive research on the pathogenesis of polycystic ovary syndrome (PCOS), there is still disagreement on the underlying mechanisms. The rat model for experimentally induced polycystic ovaries (PCO)-produced by a single injection of estradiol valerate-has similarities with human PCOS, and both are associated with hyperactivity in the sympathetic nervous system. Nerve growth factor (NGF) is known to serve as a neurotrophin for both the sympathetic and the sensory nervous systems and to enhance the activity of catecholaminergic and possibly other neuron types. Electro-acupuncture (EA) is known to reduce hyperactivity in the sympathetic nervous system. For these reasons, the model was used in the present study to investigate the effects of EA (12 treatments, approximately 25 min each, over 30 days) by analyzing NGF in the central nervous system and the endocrine organs, including the ovaries. The main findings in the present study were first, that significantly higher concentrations of NGF were found in the ovaries and the adrenal glands in the rats in the PCO model than in the control rats that were only injected with the vehicle (oil or NaCl). Second, that repeated EA treatments in PCO rats resulted in concentrations of NGF in the ovaries that were significantly lower than those in non-EA-treated PCO rats but were within a normal range that did not differ from those in the untreated oil and NaCl control groups. The results in the present study provide support for the theory that EA inhibits hyperactivity in the sympathetic nervous system.

Effects of electro-acupuncture on anovulation in women with polycystic ovary syndrome.

BACKGROUND: The present study was designed to evaluate if electro-acupuncture (EA) could affect oligo-/anovulation and related endocrine and neuroendocrine parameters in women with polycystic ovary syndrome (PCOS). METHODS: Twenty-four women (between the ages of 24 and 40 years) with PCOS and oligo-/amenorrhea were included in this non-randomized, longitudinal, prospective study. The study period was defined as the period extending from 3 months before the first EA treatment, to 3 months after the last EA treatment (10-14 treatments), in total 8-9 months. The menstrual and ovulation patterns were confirmed by recording of vaginal bleedings and by daily registrations of the basal body temperature (BBT). Blood samples were collected within a week before the first EA, within a week after the last EA and 3 months after EA. RESULTS: Nine women (38%) experienced a good effect. They displayed a mean of 0.66 ovulations/woman and month in the period during and after the EA period compared to a mean of 0.15 before the EA period (p=0.004). Before EA, women with a good effect had a significantly lower body-mass index (BMI) (p<0.001), waist-to-hip circumference ratio (WHR) (p=0.0058), serum testosterone concentration (p=0.0098), serum testosterone/sex hormone binding globulin (SHBG) ratio (p=0.011) and serum basal insulin concentration (p=0.0054), and a significantly higher concentration of serum SHBG (p=0.040) than did those women with no effect. CONCLUSION: Repeated EA treatments induce regular ovulations in more than one third of the women with PCOS. The group of women with good effect had a less androgenic hormonal profile before treatment and a less pronounced metabolic disturbance compared with the group with no effect. For this selected group EA offers an alternative to pharmacological ovulation induction.

Long-term follow-up of patients treated with acupuncture for xerostomia and the influence of additional treatment.

OBJECTIVE: To determine the long-term effects of acupuncture in patients with xerostomia of different etiologies and the influence of additional treatment. DESIGN: Retrospective study. SUBJECTS: Seventy patients, between the ages of 33 and 82, with xerostomia due to primary and secondary Sjögren's syndrome, irradiation and other causes were included. The median duration of xerostomia was 32 months. METHODS: Salivary flow rates (SFR) for whole unstimulated and stimulated saliva were used as indicators of effects of treatment. Data from 67/70 patients were analyzed 6 months following a baseline course of 24 acupuncture treatments using two-way ANOVA. Patients data up to 3 years were also compared by those who chose to receive additional acupuncture treatment vs those who did not. These data were analyzed descriptively. RESULTS: Statistically significant differences in unstimulated and stimulated salivary flow rates (P < 0.01) were found in all etiological groups after 24 acupuncture treatments and up to 6 months follow-up compared to baseline. Three years observation of these patients showed that patients receiving additional acupuncture treatment had a consistently higher median SFR in both unstimulated and stimulated saliva compared to patients who chose not to continue acupuncture. The upper limits of the interquartile range were also higher. CONCLUSIONS: This study shows that acupuncture treatment results in statistically significant improvements in SFR in patients with xerostomia up to 6 months. It suggests that additional acupuncture therapy can maintain this improvement in SFR for up to 3 years.

Sensory stimulation (massage) reduces blood pressure in unanaesthetized rats.

The objective of this study was to investigate how sensory stimulation by massage-like stroking influences blood pressure and heart rate in conscious rats. Also, the influence of different locations and durations of the stimulation were assessed. For this purpose, the ventral side of the abdomen or the dorsal side of the back was manually stroked at a speed of approximately 20 cm/s, with a frequency of 0.67 Hz and at an estimated pressure of 100 mm H2O. During the treatment, the rats were held across the scapula and the neck region. Blood pressure and heart rate were measured with the cuff technique before treatment and repeatedly during the post-stimulatory period. Massage-like stroking for 5 min of the abdominal area produced a maximum decrease of approximately 20 mm Hg in blood pressure and 60 beats/min in heart rate. This reduction remained significant at 3 and 4 h after stimulation, respectively. Stimulation of the abdominal area for 2 min produced a less pronounced decrease in blood pressure as compared to the 5-min stroking. Stroking of the back resulted in a short-lasting blood pressure increase that gradually returned to the baseline level within the post-stimulatory observation time. Control animals that were handled in the same way as the experimental animals except for the stroking showed an increase of approximately 20 mm Hg in blood pressure and 60 beats/min for about 1 h after the cessation of the handling. The responses of the blood pressure and heart rate to both abdominal and back massage were significantly inhibited as compared to the control animals. These results suggest that massage-like stroking of the skin produces an inhibitory effect on the cardiovascular excitatory responses in rats. Especially, the results of the present study demonstrate that massage-like stroking of the abdomen reduces both blood pressure and heart rate below the pre-stimulus baseline levels.