Nisaudah Radenahmad

Accumulation of calbindin in cortical pyramidal cells with ageing; a putative protective mechanism which fails in Alzheimer's disease

There is considerable interest in the status of calbindin immunoreactive neurones in Alzheimers disease (AD) but previous studies have produced widely differing results. Here we describe calbindin neurones in temporal neocortex from 18 severely demented patients with neuropathologically confirmed AD and 13 age and post‐mortem delay matched, neurologically normal controls. Calbindin immunoreactive neurones were small and round in layers II–IV, and pyramidal in layers IIIc and V. There were significantly more calbindin positive neurones in controls than in AD (mean± SD, for each comparison P < 0.01): superior temporal lobe, AD = 3.32 ± 2.24, Control (C) = 24.83 ± 10.8; middle temporal lobe, AD = 3.6 ± 4.94, C = 26.09 ± 15.7; inferior temporal lobe, AD 3.69 ± 3.6, C = 25.25 ± 16.9. Furthermore, there was an age‐related increase in immunopositive neurones in the superior (r2 = 0.37, P = 0.046) and inferior (r2 = 0.75, P = 0.01) temporal gyri in controls. In AD the number of calbindin positive neurones did not change with age. This is the first report of such an age‐related increase in controls, and it suggests that this, rather than a decrease in AD, accounts for the overall difference between AD and controls. It is possible that an increase in intraneuronal calbindin protects these cells from degeneration and that failure of such a neuroprotective mechanism is a significant contributory factor in the pathogenesis of AD.

Effects of Kaempferia parviflora rhizomes dichloromethane extract on vascular functions in middle-aged male rat.

ETHNOPHARMACOLOGICAL RELEVANCE In Thai traditional medicine, rhizomes of Kaempferia parviflora (KP) have been used for treating hypertension and for the promotion of longevity with good health and well being. Ageing is one of the most important risk factors for development of cardiovascular disease. To investigate whether a 6 weeks oral administration of a dichloromethane extract of fresh rhizomes of Kaempferia parviflora (KPD) had any effects on vascular functions, on the accumulation of lipid, as well as on any signs of gross organ toxicity in middle-aged rats. MATERIALS AND METHODS Fresh rhizomes of Kaempferia parviflora were first macerated twice with 95% ethanol to remove the dark color before extracting three times with 100% dichloromethane. The dichloromethane extract was evaporated under reduced pressure to obtain the dried Kaempferia parviflora dichloromethane extract (KPD). The rats were orally administered with the KPD at a dosage of 100mg/kg body weight, or with the same volume of the vehicle (tween 80, 0.2g: carboxy-methylcellulose sodium, 0.2g: distilled water 10 ml) once or twice a day for 6 weeks. Vascular functions were studied on isolated thoracic aorta and the mesenteric artery. The vascular eNOS enzyme was measured by Western blot analysis. Blood chemistry was measured by enzymatic methods. Liver cell lipid accumulation was measured using oil red O staining. RESULTS A 6 weeks treatment of KPD once a day had no significant effects on any of the studied parameters. When the KPD was given twice a day, the contractile responses to phenylephrine of the thoracic aorta and mesenteric artery were lower than the vehicle control group, and this effect was abolished by N(G)-nitro-l-arginine or by removal of the vascular endothelium. Vasorelaxation to acetylcholine, but not to glyceryl trinitrate, by the thoracic aortic and mesenteric ring precontracted with phenylephrine was higher from the KPD treated rats than those from the vehicle control groups. Western blot analysis showed a higher quantity of thoracic- and mesenteric-eNOS protein obtained from the KPD treated rats. In addition, the body weight, serum glucose and triglycerides levels, visceral and subcutaneous fat, as well as liver lipid accumulation were all significantly decreased in the KPD treated rats compared to those of the vehicle control. No differences were found between the KPD treated-, and the vehicle-control for animal food intake, internal organ weight, serum ALP, SGOT, SGPT, BUN and creatinine levels, serum cholesterol, HDL-C and LDL-C levels, nor total blood cell counts. CONCLUSIONS The chronic oral administration of KPD extract, to middle aged rats, caused a decrease in vascular responsiveness to phenylephrine with an increase in the acetylcholine induced vasorelaxation, due to an increase in nitric oxide production from their blood vessels. The extract also caused a decrease in visceral and subcutaneous fat, fasting serum glucose and triglyceride levels and liver lipid accumulation, with no changes to liver and kidney functions or to total blood cell counts. It is possible that these KPD extracts could be developed as a health product for mid-aged humans to reduce obesity, diabetes type II and cardiovascular disease.

Six weeks oral gavage of a Phyllanthus acidus leaf water extract decreased visceral fat, the serum lipid profile and liver lipid accumulation in middle-aged male rats

ETHNOPHARMACOLOGICAL RELEVANCE Advancing age is associated with an increased accumulation of visceral fat and liver lipid which is then responsible for an age-related risk for cardiovascular disease. Looking after ourselves well with suitable micronutrients could prevent disease or prolong our healthy cardiovascular functions. In Thai traditional medicine, leaves of Phyllanthus acidus (PA) have been used for many purposes including as an antihypertensive agent and to provide relief from a headache caused by hypertension. We aimed to investigate the effects of a chronic oral administration of PA extracts to middle-aged (12-14 months) rats on their body weight, food intake, body fats, liver and kidney functions, fasting blood glucose and lipid profiles, liver lipid accumulation and on blood pressure. MATERIALS AND METHODS Three different kinds of PA extracts were used: (1) a PA water extract, (2) a heated PA water extract, and (3) an n-butanol fraction of the PA water extract, prepared from fresh leaves of Phyllanthus acidus. The rats were orally gavaged with the three PA extracts at 1.0 g/kg body weight or, as a control, with distilled water once a day for 6 weeks. Fasting blood sugar, lipid profile and ALP, SGOT, SGPT, BUN and creatinine levels were measured by enzymatic methods. Liver lipid accumulation was measured using oil red O staining on fresh thin cryostat liver tissue sections. The animal basal blood pressure and heart rate were measured in anesthetized rats via a common carotid artery using a polygraph. RESULTS Results showed that after 6 weeks of treatment using gavaged heated PA extract and PA n-butanol extract there were no changes in any of the parameters studied. However, the initial PA water extract caused a slight decrease in the animal body weight with no change in food intake. No changes were observed in the liver and kidney functions (serum ALP, SGOT, SGPT, BUN and creatinine did not change), nor did the fasting blood sugar or triglyceride levels differ significantly. Serum cholesterol, HDL and LDL levels, as well as visceral and subcutaneous adipose tissue and liver lipid accumulation were significantly decreased compared to that of the control group. There were no differences found in the basal systolic and diastolic blood pressure and the basal heart rate between the PA water extract treatment and the control group. CONCLUSIONS These results indicated that the PA water extract had an effect on lipid metabolisms that resulted in a decrease of the serum lipid profile, visceral and subcutaneous fat, as well as on liver lipid accumulation in middle-aged rats. The active component that is responsible for these effects is likely to be a water soluble substance(s) and is heat labile. As a consequence of these beneficial effects of the PA water extract, it would be a good choice for further development for use as a nutraceutical or health product to prevent and/or to slow down the development of obesity and/or cardiovascular disease.

Young coconut juice can accelerate the healing process of cutaneous wounds

BackgroundEstrogen has been reported to accelerate cutaneous wound healing. This research studies the effect of young coconut juice (YCJ), presumably containing estrogen-like substances, on cutaneous wound healing in ovairectomized rats.MethodsFour groups of female rats (6 in each group) were included in this study. These included sham-operated, ovariectomized (ovx), ovx receiving estradiol benzoate (EB) injections intraperitoneally, and ovx receiving YCJ orally. Two equidistant 1-cm full-thickness skin incisional wounds were made two weeks after ovariectomy. The rats were sacrificed at the end of the third and the fourth week of the study, and their serum estradiol (E2) level was measured by chemiluminescent immunoassay. The skin was excised and examined in histological sections stained with H&E, and immunostained using anti-estrogen receptor (ER-α an ER-β) antibodies.ResultsWound healing was accelerated in ovx rats receiving YCJ, as compared to controls. This was associated with significantly higher density of immunostaining for ER-α an ER-β in keratinocytes, fibroblasts, white blood cells, fat cells, sebaceous gland, skeletal muscles, and hair shafts and follicles. This was also associated with thicker epidermis and dermis, but with thinner hypodermis. In addition, the number and size of immunoreactive hair follicles for both ER-α and ER-β were the highest in the ovx+YCJ group, as compared to the ovx+EB group.ConclusionsThis study demonstrates that YCJ has estrogen-like characteristics, which in turn seem to have beneficial effects on cutaneous wound healing.

Angiotensin II may mediate apoptosis via AT1-receptors in the rat cardiac conduction system

Introduction Apoptosis and angiotensin II (Ang II) have been suggested as possible causes of arrhythmias. In addition, Ang II via Ang II type I (AT1-) receptors, has been demonstrated to induce cardiomyocyte apoptosis. The transgenic m(Ren-2)27 (TG) rat carries the additional Ren-2 gene, the expression of which results in an increase in cardiac Ang II, thus potentially affecting the cell growth/death equilibrium. In this study we have investigated the effect of Ang II, via AT1-receptors, on mediating apoptosis in a cardiac conduction system (SA node and AV nodes). Materials and methods Heart sections from male two-day, one-week and two-week TG and Sprague-Dawley (SD) rats were stained with Masson Trichrome to localise the SA and AV nodes. The sections containing SA or AV nodes were processed for quantitation of apoptotic nuclei and AT1-receptors. Results The number of apoptotic nuclei/mm2 in the SA and AV nodes were found to decrease from two days to two weeks in both the TG and the SD rats, and the number of apoptotic nuclei/mm2 in the TG groups was significantly higher than that of the SD groups for all ages (p<0.05). The number of AT1-receptors/mm2 in the SA node were found to decrease with increasing age, whereas the number of AT1-receptors/mm 2 in the AV node was increased in both TG and SD rats and the number of AT1-receptors/mm2 in the three TG groups was significantly more than that of the three SD groups (p<0.05). Discussion and conclusion As a consequence of the additional renin gene in the TG rats, which results in the alteration of the local renin-angiotensin system, the numbers of AT1-receptors/mm 2 and apoptotic nuclei/mm2 are increased. The number of apoptotic nuclei/mm2 and AT1-receptors/mm2 in the SA node decrease with maturation, whereas, the number of AT1-receptors in the AV node increase. Thus, there may be a correlation between Ang II and apoptosis in the SA node, which does not appear to be present in the AV node.