Proton nuclear magnetic resonance analysis of meconium composition in newborns.
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Cholesterol↗
Biomedical subjects
C Zancanaro
Publications and source records attributed to C Zancanaro.
Quantitative ultrastructural changes of hepatocyte constituents in euthermic, hibernating and arousing dormice (Muscardinus avellanarius).
Hibernating animals represent a suitable model for investigating the structural effects of drastic changes in cell activity under physiological conditions. In this study we investigated by means of electron microscopy and morphometrical analysis the fine structural counterpart of functional rest in hepatocytes of the hibernating dormouse, Muscardinus avellanarius, in comparison with arousing and euthermic dormice. Our observations demonstrate that during hibernation several structural constituents of the hepatocyte undergo modifications. In particular, during deep hibernation, the total cell and cytoplasm area significantly reduced, as well as the total and percent glycogen and residual body area, and the Golgi apparatus almost disappeared. Upon arousal, the amount of glycogen was minimal, whereas total cell and cytoplasm area significantly increased towards the euthermic value as well as total and percent residual body area. In comparison with the euthermic condition, the total and percent cell lipid area significantly increased in early hibernation, reduced in deep hibernation and almost disappeared during arousal. Taken together, our findings give quantitative ultrastructural support to the marked reduction found in hepatocyte functional activities during hibernation. Such a reduced activity involves profound rearrangement of the euthermic cell structure, which is rapidly resumed upon arousal.
Animals↗
Proton and carbon NMR spectroscopic analysis of rat liver postnatal development.
BACKGROUND: In the present paper compositional changes of rat liver from birth to adult age were investigated by proton and carbon nuclear magnetic resonance (NMR) spectroscopy. METHODS: The water-soluble and organic (lipid) fraction of pooled liver tissue from rats aged 1,7, 21 days, and 5 months were extracted according to Folch's method and analysed by proton and carbon nuclear magnetic resonance spectroscopy at 500 MHz. Phospholipids separated by chromatography from the organic fraction were also analysed. RESULTS: In proton spectra of the water-soluble fraction several metabolites were identified. Developmental changes in the ratio between betaine and choline and between 3-hydroxybutyrate and several other organic acids were shown. In proton spectra of the total organic fraction, signals from fatty acids chain, phophocholine and glycerol were assigned unambiguously. Phospholipids fatty acid acyl chain length, mean unsaturation and mean polyunsaturation increased from birth to weaning. The relative amounts of unsaturated fatty acids did not show obvious changes during the investigated period. CONCLUSIONS: This study shows that NMR spectroscopy is a useful tool to investigate postnatal developmental changes in the liver chemical constituents with a minimum of preparative procedures.
Journal Article↗
Fine structural modifications of liver, pancreas and brown adipose tissue mitochondria from hibernating, arousing and euthermic dormice.
An ultrastructural and morphometric study was performed on mitochondria of euthermic, hibernating and arousing hazel dormice (Muscardinus avellanarius), in order to investigate possible modifications during the seasonal cycle. Hepatocytes, pancreatic acinar cells and brown adipocytes were considered. Our results demonstrated that: (1) the general morphology of mitochondria of all cell types shows slight modifications during the seasonal cycle; (2) mitochondrial size and inner membrane length significantly increase from euthermia to hibernation and decrease upon arousal in all cell types; (3) mitochondrial matrix granules drastically increase in number during hibernation and decrease upon arousal in hepatocytes and pancreatic acinar cells, whereas they do not change in brown adipocytes. These structural modifications are probably related to the changes in cellular energy needs during the euthermia-hibernation-arousal cycle.
Adipose Tissue, Brown↗
NMR spectroscopic analysis of rat brain development: in vitro proton and carbon studies of whole tissue and its phospholipid fraction.
The developmental patterns of the rat brain at several postnatal time points were investigated in Folch (chloroform-methanol) extracts. The chloroform- (lipid-containing) and water-soluble (cytosolic) fractions of whole-tissue extracts and the phospholipid fraction separated from the organic fraction by chromatography were analysed by means of high-resolution (1)H and (13)C nuclear magnetic resonance (NMR) spectroscopy. Analysis of the cytosolic fraction showed the changing patterns of several brain metabolites during postnatal maturation, in full agreement with data obtained from perchloric acid extracts. (1)H NMR spectroscopy of the phospholipid fraction allowed for quantitative evaluation of fatty acid acyl chain length, mean unsaturation and mean polyunsaturation. It was found that both mean unsaturation and polyunsaturation are lower in adult brain phospholipids than during the first 3 postnatal weeks. (13)C NMR spectroscopy of the same fraction showed that the molar percentage of C(18) fatty acids (oleic, linoleic, linolenic) in brain phospholipids is similar at all the investigated time points. These results indicate that the combination of Folch extraction with simple chromatographic procedures and NMR analysis yields useful data to define the chemical maturation of the brain.
Animals↗
In vivo octreotide administration acutely reduces exocrine granule size in the human pancreas.
BACKGROUND: Octreotide has been found to be effective in the prevention of postoperative complications of pancreatic surgery, but the benefit of preoperative octreotide administration has not been assessed. AIMS: To evaluate the ability of octreotide in reducing the amount of digestive enzymes in the pancreas before surgery, a morphometric ultrastructural study of the gland was undertaken in patients undergoing demolitive pancreatic surgery. METHODS: Twenty-three inpatients received saline (n = 8) or octreotide (100 micrograms s.c.) before surgery either once (n = 5), or three (n = 5) or six (n = 5) times at 8-hour intervals. At surgery, biopsies of the pancreas were taken and processed for electron microscopy. Several parameters were assessed in exocrine cells by means of ultrastructural morphometry. RESULTS: A single administration of octreotide significantly reduced the exocrine granule number and the mean and total granular surface sectional area, and the ratio between granule area and cytoplasmic area. Repeated octreotide administrations were associated with partial (3 administrations) and complete (6 administrations) recovery of all parameters to control values. CONCLUSION: Preoperative administration of octreotide, the synthetic analogue of somatostatin, acutely reduces exocrine granule number and size in the pancreatic cell. This finding can partially explain the prophylactic effect of the drug on early complications of pancreatic surgery. Such an effect is not maintained over multiple administrations of the somatostatin analogue. Possible explanations for this latter finding are discussed.
Adult↗
The calcium-channel blocker lacidipine reduces the development of atherosclerotic lesions in the apoE-deficient mouse.
BACKGROUND: Lacidipine is a widely used calcium-channel blocker, which has both long-lasting antihypertensive activity and also antioxidant properties. Previous studies have demonstrated the ability of lacidipine to reduce the development of atherosclerotic lesions in several animal models. OBJECTIVE: The present study investigated the antiatherosclerotic potential of lacidipine in the apoE-deficient mouse, an experimental model of atherosclerosis showing progressively complex and widespread lesions which closely resemble the inflammatory-fibrous plaques seen in humans. METHODS: Lacidipine was administered daily by gavage for 10 weeks at dose levels of 0 (control), 0.3, 1.0 and 3.0 mg/kg. RESULTS: Lacidipine administration reduces the extension of atherosclerotic lesions in the aorta of the apoE-deficient mouse without affecting plasma lipid levels. We also show that apoE-deficient mice have four-fold higher values of the proatherogenic peptide, endothelin, compared with the wild-type C57BL/6 mouse and that lacidipine administration reduced, in a dose-dependent manner, the concentrations of plasma endothelin. CONCLUSION: Lacidipine has anti-atherogenic effects in the apoE-deficient mouse, and reduces plasma endothelin concentrations.
Animals↗
[Facial localized cosmetic filling by multiple injections of fat stored at -30 degrees C. Techniques, clinial follow-up of 99 patients and histological examination of 10 patients].
At the end of the 1980's and at the beginning of the 1990's, free fat autografts were abandoned because of the poor long-term results. Two theories supported by hystological studies (the host replacement theory or the cell survival theory) have now been proposed to explain survival of fat tissue. They demonstrate that, when harvested by a correct technique, fat tissue presents the ideal characteristics to be grafted, providing a good soft-tissue substitute with a minimum follow-up of four years up to 6 years. The authors present a multiple stage lipofilling technique in 99 cases of facial rejuvenation by means of lipolayering. Injections are performed every 20 days and fat is stored fat at -30 degrees C with good aesthetic results and minimal morbidity with clinical follow-up at 4 and 6 years. Histological examination of fat stored for 20 days and for 8 months was also performed in 10 patients. Microscopic and ultrastructural examination demonstrated that stored adipocytes retain their anatomical features and, when injected in multiple stages, are adequate to obtain good clinical results. Our histological study combined with the clinical outcomes, suggests the advantage of several stage injection of stored fat (-30 degrees C).
Adipocytes↗
[The digestive apparatus during hibernation: morphological and functional changes].
During hibernation feeding is suspended and the digestive apparatus is in a condition of functional rest. This paper is a review of data published in the period from 1966 to 1999 on the structure/function relationship in the digestive apparatus of hibernating animals. The available information is partial, due to differences in methodology and species used; however, it is apparent from the data that the digestive apparatus undergoes an important rearrangement during hibernation, inclusive of mass loss and reduced/arrested mitotic activity, in the presence of a generally preserved morphology of the organs. All changes are reversible upon arousal. Recent findings show that, during hibernation, protein expression may be enhanced in the intestine as well as transport activities in the mucosa. Therefore, the hibernation condition is finely regulated, representing a potentially useful model for studies aimed at improving organ preservation procedures (e.g. for transplantation).
English Abstract↗
Neuropeptide expression in the mouse vomeronasal organ during postnatal development.
The expression of selected regulatory neuropeptides was investigated by immunohistochemistry in nerves supplying the vomeronasal organ (VNO) of mice during postnatal development. Results show that neurons in the VNO are devoid of immunolabeling with any of the antibody used from 1 day to 2 months of age. In the non-receptor epithelium (NRE) and the vomeronasal vascular pump (VP) the timing of expression of regulatory neuropeptides differed among neuropeptides and the different VNO structures. Regulatory neuropeptides usually found in sensory nerves (substance P, calcitonin gene-related peptide) and efferent nerves (neuropeptide Y, atrial natriuretic peptide) are expressed in the NRE and the VP, respectively. These results support the view that the VNO is to some extent functional during postnatal development.
Animals↗
Immunohistochemical investigation of the vomeronasal organ. Nitric oxide synthase expression in the mouse during postnatal development.
The expression of nitric oxide synthase type I (NOS-I), the key enzyme for the synthesis of the gaseous neurotransmitter nitric oxide, was investigated by means of immunohistochemistry in the vomeronasal organ (VNO) of mice from postnatal day 1 for 2 months. The results show that NOS is expressed in extrinsic nerve supplying the developing erectile tissue of VNO (the so-called VNO pump) as well as blood vessels in the connective tissue laying under the receptor epithelium at postnatal day 1. At 8, 15 and 21 postnatal days, and at 2 months the density of NOS-1-immunoreactive nerves goes along with the development of the erectile tissue. From postnatal day 8 onwards, NOS-1-immunoreactive fibers are found also in the vicinity of the VNO glands. These data suggest that nitric oxide (NO) modulates VNO activity early after birth in the mouse.
Age Factors↗
Nuclear bodies are usual constituents in tissues of hibernating dormice.
In previous studies we demonstrated in several tissues of the hazel dormouse Muscardinus avellanarius that during hibernation cell nuclei contain particular structural constituents absent in euthermia. In the present study we examine the same tissues in euthermic and hibernating individuals of the edible dormouse Glis glis in order to investigate possible modifications of nuclear structural constituents occurring during hibernation in this species. Edible dormice were captured in the wild and maintained in an external animal house. Samples of liver, pancreas, brown adipose tissue and adrenal cortex were taken from three hibernating and three euthermic animals and processed for resin embedding. Ultrastructural and immunocytochemical studies were carried out on cell nuclei of these tissues. The most evident feature of cell nuclei of hibernating dormice was the presence of several nuclear bodies, namely fibro-granular material, amorphous bodies, coiled bodies, perichromatin granule-like granules and nucleoplasmic fibrils, the distribution of which was peculiar to each tissue. No one of these constituents was detectable during euthermia. Immunocytochemical analyses revealed that they contain some splicing factors. Apart from some differences, maybe due to the different characteristics of lethargy, the nuclear bodies found in edible dormice were morphologically and immunocytochemically similar to those previously described in the same tissues of hazel dormice. They therefore seem to be strictly correlated to the hibernating state. If they represent storage and/or assembly sites of splicing factors to be rapidly used upon arousal, they could represent a usual structural feature in cells of hibernating species.
Adipose Tissue, Brown↗
alpha-Gustducin expression in the vomeronasal organ of the mouse.
The expression of alpha-gustducin, a G protein alpha subunit involved in bitter and sweet taste transduction, was investigated in chemosensory tissues of adult mice. By immunohistochemistry, alpha gustducin was absent in the olfactory neuroepithelium. Instead, alpha gustducin was expressed in a subset of bipolar cells in the proliferative zone of the vomeronasal neuroepithelium as well as in taste buds. Northern blot analysis confirmed the presence of alpha gustducin in isolated vomeronasal organs. Moreover, immunohisto- chemistry revealed the expression of alpha gustducin in scattered cells of the nasal respiratory epithelium. These results show for the first time that alpha gustducin is expressed in chemosensory tissue outside the alimentary tract, suggesting that common transduction mechanisms could be shared by apparently unrelated chemosensory tissues.
Animals↗
Immunohistochemical evidence suggests intrinsic regulatory activity of human eccrine sweat glands.
Immunohistochemistry of normal eccrine sweat glands was performed on paraffin sections of human skin. Immunoreactivity (ir) for neuron specific enolase, S100 protein (S100), regulatory peptides, nitric oxide synthase type I (NOS-I) and choline-acetyltransferase (ChAT) was found in small nerve bundles close to sweat glands. In the glands, secretory cells were labelled with anticytokeratin antibody. Using antibodies to S100, calcitonin gene-related peptide (CGRP) and substance P (SP) a specific distribution pattern was found in secretory cells. Granulated (dark) and parietal (clear) cells were immunopositive for CGRP, and S100 and SP, respectively. Immunoreactivity was diffuse in the cytoplasm for CGRP and S100, and peripheral for SP. Myoepithelial cells were not labelled. Electron microscopy revealed electron dense granules, probably containing peptide, in granulated cells. Using antibodies to NOS-I and ChAT, ir was exclusively found in myoepithelial cells. Immunoreactivity for the atrial natriuretic peptide was absent in sweat glands. These results provide evidence for the presence of both regulatory peptides involved in vasodilation and key enzymes for the synthesis of nitric oxide and acetylcholine in the secretory coil of human sweat glands. It is suggested that human sweat glands are capable of some intrinsic regulation in addition to that carried out by their nerve supply.
Adult↗
The kidney during hibernation and arousal from hibernation. A natural model of organ preservation during cold ischaemia and reperfusion.
BACKGROUND: During hibernation the kidney is in a hypothermic condition where renal blood flow is minimal and urine production is much reduced. Periodical arousal from hibernation is associated with kidney reperfusion at increasing body temperature, and restored urine production rate. METHODS: To assess the degree of structural preservation during such extreme conditions, the kidney cortex was investigated by means of electron microscopy in the dormouse Muscardinus avellanarius during winter hibernation, arousal from hibernation and the summer active period. RESULTS: Results show that the fine structure of the kidney cortex is well preserved during hibernation. In the renal corpuscle, a sign of slight lesion was the focal presence of oedematous endothelial cells and/or podocytes. Proximal convoluted tubule cells showed fully preserved ultrastructure and polarity, and hypertrophic apical endocytic apparatus. Structural changes were associated with increased plasma electrolytes, creatinine and urea nitrogen, and proteinuria. During the process of arousal the fine structure of the kidney cortex was also well maintained. CONCLUSION: These results demonstrate that dormice are able to fully preserve kidney cortex structure under extreme conditions resembling e.g. severe ischaemia or hypothermic organ storage for transplantation, and reperfusion. Elucidation of the mechanisms involved in such a natural model of organ preservation could be relevant to human medicine.
Animals↗
Ultrastructural, morphometrical and immunocytochemical analyses of the exocrine pancreas in a hibernating dormouse.
Pancreatic acinar cells of euthermic, hibernating and arousing individuals of the hazel dormouse Muscardinus avellanarius (Gliridae) have been observed at the electron-microscopic level and analysed by means of ultrastructural morphometry and immunocytochemistry in order to investigate possible fine structural changes of cellular components during periods of strikingly different degrees of metabolic activity. During hibernation, the cisternae of the rough endoplasmic reticulum (RER) flatten assuming a parallel pattern, the Golgi apparatus is extremely reduced and the mitochondria contain many electron-dense particles. The cell nuclei appear irregularly shaped, with deep indentations containing small zymogen granules. They also contain abundant coiled bodies and unusual constituents, such as amorphous bodies and dense granular bodies. Large numbers of zymogen granules occur in all animals. However, the acinar lumina are open and filled with zymogen only in euthermic animals, whereas, in hibernating and arousing individuals, they appear to be closed. Morphometrical analyses indicate that, in pancreatic acinar cells, nuclei and zymogen granules significantly decrease in size from euthermia to hibernation, probably reflecting a drastic decrease of metabolic activities, mainly protein synthesis and processing. In all the studied animals, immunocytochemistry with specific antibodies has revealed an increasing gradient in alpha-amylase content along the RER-Golgi-zymogen granule pathway, reflecting the protein concentration along the secretory pathway. Moreover, during deep hibernation, significantly larger amounts of alpha-amylase accumulate in RER and zymogen granules in comparison to the other seasonal phases analysed. Upon arousal, all cytoplasmic and nuclear constituents restore their euthermic aspect and all morphometrical and immunocytochemical parameters exhibit the euthermic values, thereby indicating a rapid resumption of metabolic activities.
Amylases↗
Carcinomalike monotypic epithelioid angiomyolipoma in patients without evidence of tuberous sclerosis: a clinicopathologic and genetic study.
We report the clinicopathologic, immunohistochemical, ultrastructural, and genetic features of an unusual renal tumor composed of large, atypical, densely packed, clear/eosinophilic epithelioid cells. Three patients, two men and one woman (ages 31, 36, and 60 years of age, respectively), had abdominal pain. Morphologically, all cases showed aggressive features (largeness, atypical cells, sarcomatoid features, necrosis, and, in one case, invasion of the renal vein). Despite the marked morphologic resemblance of these tumors to high-grade sarcomatoid renal cell carcinoma, their phenotype (HMB45+, CD68+/-, actin+/-, and vimentin and keratin negative) is in contrast to that observed in epithelial tumors and parallels the phenotypic profile of angiomyolipoma. Ultrastructural analysis showed the presence of glycogen, mitochondria, and prominent electron-dense, membrane-bound granules in the neoplastic cells, and the absence of melanosomes or premelanosomes. Genetic study, performed using polymerase chain reaction from paraffin sections, showed a loss of heterozygosity at the TSC2-containing region on 16p in one case, and on 3p in two cases, showing that multiple genetic alterations are taking place in these tumors. Follow-up has shown local recurrence in one case after 6 years, and the patient died 1 year later of cardiorespiratory failure. The other two patients are well after 26 and 10 months. All three patients were evaluated for signs of tuberous sclerosis, and findings were negative. We suggest that these tumors should be considered close relatives of the angiomyolipoma variants, composed purely of perivascular epithelioid cells. More cases and longer follow-up durations are needed to fully evaluate its prognostic implication.
Adult↗
Ultrastructure of the adrenal cortex of hibernating, arousing, and euthermic dormouse, Muscardinus avellanarius.
BACKGROUND: The adrenal gland is a key organ for hibernation (a condition characterized by striking reduction of body functions). Very limited information is available on the fine structure of the gland during hibernation and on the periodical arousal from hibernation. METHODS: Dormice (Muscardinus avellanarius) were maintained in an external animal house and allowed to hibernate spontaneously (November). Arousal was induced in March by exposure to daylight. Euthermic, active dormice were captured in June. The adrenals were taken from four hibernating, three arousing, and four euthermic dormice and processed for resin embedding. The ultrastructure of the adrenal cortex was investigated by transmission electron microscopy. RESULTS: In the zona glomerulosa of hibernating and arousing dormice, the smooth endoplasmic reticulum was prominent in comparison with euthermic animals, and mitochondria showed abundant vesicular cristae. The zona fasciculata and zona reticularis did not show consistent differences, apart from a lower cell lipid content in the outer portion of zona fasciculata of arousing dormice. CONCLUSIONS: The zona glomerulosa showed signs of increased activity during hibernation. This finding is supported by previous biochemical data demonstrating increased production of renin and aldosterone during such extreme physiological conditions. Activation of the zona glomerulosa in hibernation is probably adaptive to a condition of drastically reduced salt intake.
Adrenal Cortex↗