Age-Related Features of Cataractogenesis in Salmon Fry. 2. Biochemical Features of Lens during Cataractogenesis

We studied biochemical features of the lipid and phospholipid composition and patter of fatty acid spectra in lenses of one and two year old salmons bearing cataracts. The lipid complex of lenses of diseased fish underwent significant changes. Cataractogenesis was accompanied by enhanced free radical oxidation and accumulation of malone dialdehyde in lenses of salmons of various ages. The intensification of oxidative processes was synchronized with the decreased level of antioxidant protection (reduced glutathione, vitamins A and E). The activity of Ca²⁺-dependent proteolytic enzymes in lenses was determined and its decrease in the case of cataract was shown.     

Age-Related Features of Cataractogenesis in the Salmon Fry. 1. Lipid Composition of Lens during Normal Development

This papers opens a series of publications on the mechanisms of cataractogenesis in the salmon fry. Biochemical features of normal lens development and cataractogenesis in fry of different age and age-related dynamics of the liver lipid composition during upon cataractogenesis will be dealt with, since lipids are most intensely synthesized in the liver, from where they are transported in the lens. Here, we describe the dynamics of lens lipid composition in the salmon fry, including the total lipid content and dynamics of individual classes. The data are analyzed on the fatty acid spectrum of the salmon fry lens, as compared to the human lens.     

Age-Related Features of Ceruloplasmin Biosynthesis and Distribution in Rats

In vivo biosynthesis of ceruloplasmin (Cp), a copper-containing glycoprotein, which plays an important role in copper transfer between organs and bidirectional iron transport in vertebrates, was studied in 7-day old rats, which are characterized by the embryonic type of copper metabolism. In addition to the liver, Cp is synthesized in the lungs, brain, and kidneys. In pulse-chase experiments it was demonstrated that [¹⁴C]-Cp appearance in the blood coincides with the secretion ofde novo synthesized Cp from the liver. [¹⁴C]-Cp is taken up from the blood stream by cells of the heart, lung, and kidneys and binds to red blood cells, while Cp polypeptide chain is not taken up by the brain cells. Immunoreactive polypeptides of the Cp receptor were found using immunoblotting in plasma membranes of the heart, liver, kidneys, and red blood cells, rather than in the brain. Using the RT-PCR method with selective primers, it was shown that these cells contain molecular forms of Cp-mRNAs programming the synthesis of both secretory Cp and Cp bound to the plasma membrane via a glycosyl phosphatidylinositol anchor. After switching to the adult type of copper metabolism, the blood serum contents of copper and Cp sharply increase, while the Cp content in the cerebrospinal fluid, as measured according to the oxidase and antigen activities, and copper concentration, as determined by atomic absorption spectroscopy, remain low. Ontogenetic features of the system ensuring the copper homeostasis in mammals are discussed.     

Age-related features of cataractogenesis in salmon fry. 3. Age-related dynamics of liver lipid composition during cataractogenesis

When studying the cataract pathogenesis in salmon fry, we found changes in the content of individual; phospholipid fractions and fatty acid composition in the liver of diseased and healthy fish. The age-related changes correlated with the increased antioxidant activity and decreased liver content of malondialdehyde.     

Age-Related Changes in Collagen and Glycosaminoglycans of Rat Gingiva1

Age-related changes in collagen and glycosaminoglycans (GAGs) of rat gingiva were studied biochemically and histologically. The components of isolated GAGs were identified as dermatan sulphate, hyaluronic acid, and heparan sulphate. In histological studies, hyaluronic acid was present in all regions of the gingiva, whereas dermatan sulphate and heparan sulphate were present only in gingival connective tissue. However, there was no significant difference in the relationship between aging, and the content or distribution of GAGs. On the other hand, histological findings showed that collagen fibers were markedly increased in number and the vascular composition was decreased with increasing age. In biochemical studies, the content of collagen, especially of insoluble collagen, was greatly increased with age, whereas collagen biosynthesis and collagenolytic activity were markedly decreased. In addition, lysyl oxidase activity was also significantly decreased with age. The results indicate that the rate of collagen turnover decreases and collagen fibers increase in stability in rat gingiva with increasing age. These observed age-related changes may affect the ability of gingiva to respond to local irritants.     

The importance of liver in normal and silicotic lung-lipid homeostasis: 3. Triacylglycerols.

Early lung silicosis in rats activates the liver for triacylglycerol (TG) biosynthesis. The newly formed liver TG seems mobilized to the silicotic lung, as indicated by increased lung TG content and specific activity. These TG changes are associated with decreased TG content and increased TG specific activity in the liver. Serum TGs are also decreased at the study times of 6, 24, 72, and 144 h post intrapulmonary silica treatment.     

Effect of triiodothyronine on the content of phospholipids in the rat liver nuclei.

The aim of the present study was to examine the effect of triiodothyronine (T3) on the content of phospholipids and on the incorporation of blood-borne palmitic acid into the phospholipid moieties in the nuclei of the rat liver. T3 was administered daily for 7 days, 10 microg x 100 g(-1). The control rats were treated with saline. Each rat received 14C-palmitic acid, intravenously suspended in serum. 30 min after administration of the label, samples of the liver were taken. The nuclei were isolated in sucrose gradient. Phospholipids were extracted from the nuclei fraction and from the liver homogenate. They were separated into the following fractions: sphingomyelin, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine and cardiolipin. The content and radioactivity of each fraction was measured. It was found that treatment with T3 reduced the content of phosphatidylinositol and increased the content of cardiolipin in the nuclear fraction. In the liver homogenate, the content of phosphatidylinositol decreased and the content of phosphatidylethanolamine and cardiolipin increased after treatment with T3. The total content of phospholipids after treatment with T3 remained unchanged, both in the nuclear fraction and in the liver homogenate. T3 reduced the specific activity of phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine and cardiolipin and had no effect on the specific activity of sphingomyelin and phosphatidylinositol both in the fraction of the nuclei and the liver homogenate. It is concluded that excess of triiodothyronine affects the content of phospholipids in the nuclei. The changes in the content of phospholipids in the nuclei largely reflect changes in their content in the liver.     

Cytophotometric study of nuclear proteins during gametogenesis in Ascaris lumbricoides.

Reduction in the number of nucleoli/nucleus and increase in their size were usually observed in rat liver after partial hepatectomy. These changes of nucleoli were greatest 16–18 h after the operation, when RNA biosynthesis in the nucleoli is reported to be highest. Approx. 50% of the nuclei had one enlarged nucleolus at this time but after the increase in nuclear DNA synthesis less than 15% of the nuclei had one nucleolus, as in normal liver. Before the next peak of nuclear DNA synthesis, nucleolar changes appeared again, though less conspicuously.The enlarged nucleoli of regenerating liver were separated from smaller ones by discontinuous sucrose gradient centrifugation and the contents of nucleic acid and ribosomal cistrons in different-sized nucleoli were measured. The large nucleoli in regenerating liver were found to have increased DNA content, whereas smaller ones had the normal content. The total number of ribosomal cistrons in the enlarged nucleoli from regenerating liver was also increased roughly in proportion to the DNA content. No significant difference was found between the percentages of ribosomal cistrons in whole nuclear DNAs from regenerating and normal liver. Small but reproducible [³H]TdR incorporation into nucleolar DNA was observed and this was similar in normal liver and regenerating liver 12 h after partial hepatectomy. Therefore, the nucleolar changes in regenerating liver were not accompanied by any particular DNA synthesis in the nucleolus itself. These results suggest that in the nuclei of regenerating liver nucleolar chromatins may be redistributed and assembled into large nucleoli, rather than that any amplification of ribosomal cistrons occurs.     

Catecholamine concentration in rat liver after high level transection of the spinal cord.

The high level transection of the spinal cord (C-7) provokes a sustained increase of rat liver catecholamines: biphasic increase in norepinephrine level 1 hour and 24 hour after the operation and 7-fold increase of dopamine content 4 hour after the chordotomy. In contrast to cervical transection, sham operation causes only an initial catecholamine increase, the maximum being at the first hour after the surgery. Our experimental data indicate a possible participation of cervical spinal pathways in regulation of liver catecholamine content. It is also shown that bilateral adrenalectomy augments liver norepinephrine concentration in spinal rats as compared to the non-adrenalectomized ones. The results presented here indicate that cervical chordotomy affects the functioning of the sympatho-adrenal system, thus provoking specific changes in liver catecholamine content. The potential effect of such changes on a liver metabolic system (tyrosine aminotransferase induction) is discussed.     

Hepatoprotective effect of selenium and vitamin E in rabbits fed a high-fat diet.

The high-fat diet consisted of cholesterol, hydrogenated coconut oil, and cholic acid. In the blood serum and in liver homogenate lipid content, cholesterol, and triglycerides were assayed. Cytochrome P-450 concentration in liver microsomes was also estimated. In animals receiving selenium and vitamin E, the content of lipid fractions in the blood serum and liver homogenate fell, while the cytochrome P-450 content in the liver microsomes was markedly elevated. The intensified protective effect of vitamin E and selenium applied in combination against changes induced in the liver of animals receiving a HFD was confirmed by macroscopic and microscopic examination of the organ.     

Developmental and Age-Related Changes in Rat Brain Glycosaminoglycans

The quantities of each major class of glycosaminoglycan were determined in rat cerebrum from postnatal day 5 to 30 months of age. Chondroitin sulphate, dermatan sulphate, heparan sulphate, heparin, and hyaluronate were found, but no keratan sulphate was detected. Large and rapid changes in glycosaminoglycan content were observed during the period of brain maturation, and thereafter relatively steady levels were maintained until after the age of 12 months. The most remarkable change in the aged rat cerebrum was the ratio by weight of hyaluronate to chondroitin sulphate, which was approximately 1:1 from postnatal day 10 to 18 months but increased to 2.6:1 by the age of 30 months. In immature rats, the proportion of nonsulphated and 6-sulphated disaccharides derived from chondroitinase AC digests of brain glycosaminoglycans was much greater than in adults. In mature rats, chondroitin sulphate was composed almost entirely of 4-sulphated disaccharide subunits. The possibility that these changes could affect the permeability properties of the cerebral extracellular space and ionic equilibria in the brain is discussed.     

The activity of Golgi-membrane galactosyltransferase in the liver of streptozotocin-diabetic rats.

The Golgi-rich membrane fraction isolated from streptozotocin-diabetic rat liver had a lower protein content than the corresponding fraction from normal liver. Its UDPgalactose-N-acetylglucosamine galactosyltransferase activity calculated per 1 g of liver or whole liver was decreased. The electron-microscopic examination of the negatively stained fraction revealed morphological changes. The morphology of the Golgi complex in thin sections of diabetic liver was also changed.     

Age-Related Decrease of the Phosphorus Content in the Ligamentum Capitis Femoris of Monkeys

To elucidate compositional changes of the ligament with aging, the authors investigated age-related changes of elements in the ligamentum capitis femoris (LCF) of monkeys with a wide range of ages by direct chemical analysis. Used rhesus and Japanese monkeys consisted of 9 males and 22 females, ranging in age from newborn to 31 years (average age?=?10.4?±?10.9 years). After incineration with nitric acid and perchloric acid, element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that the P content decreased significantly in the LCFs of monkeys with aging, but other six element contents, Ca, S, Mg, Zn, Fe, and Na, did not change significantly with aging. Assuming that the P content indicated the active cell density and the S content indicated the protein amount, an age-related change of the mass ratio of P/S was examined in the LCFs. The mass ratio of P/S decreased significantly in the LCFs in childhood. Regarding the relationships among elements, significant direct correlations were found among the Ca, P, S, and Mg contents in the LCFs. It was suggested that the active cell density of the connective tissue cells might decrease significantly in the LCF in childhood.     

Effect of carbon-tetrachloride and mercuric chloride on the tocopherol and ubiquinone content of liver and kidney.

Authors examined changes in the total ubiquinone and tocopherol contents of livers and kidneys obtained from rats acutely poisoned by carbon-tetrachloride and mercuric-chloride. Following administration of carbon-tetrachloride after 48 h the liver ubiquinone content per 100 g tissue decreased from 11.80 +/- 2.44 mg to 7.24 +/- 2.15 mg, while the kidney ubiquinone content changed from 14.16 +/- 3.96 mg to 5.98 +/- 2.93 mg. The total tocopherol content remained unaltered. Fourty-eight hours after administration of mercuric-chloride, the liver total tocopherol and total ubiquinone contents showed no changes, the tocopherol content of the kidney dropped from 6.16 +/- 2.10 mg to 3.16 +/- 1.48 mg, while the total ubiquinone content remained unaltered.     

The Psychology of Age-Related Crises

Reform of the general education and vocational school is bringing about an improvement in the content of education, as reflected in curricula and syllabuses, textbooks, and teaching aids; teaching methods are changing, and overburdening of pupils is being eliminated.     

Connexins in Lens Development and Cataractogenesis

The lens is an avascular organ that transmits and focuses light images onto the retina. Intercellular gap junction channels, formed by at least three different connexin protein subunits, α1 (connexin43 or Gja1), α3 (connexin46 or Gja3) and α8 (connexin50 or Gja8), are utilized to transport metabolites, ions and water in the lens. In combination with physiological and biochemical analyses, recent genetic studies have significantly improved our understanding about the roles of diverse gap junction channels formed by α3 and α8 connexin subunits during lens development and cataract formation. These studies have demonstrated that α3 connexin is essential for lens transparency while α8 connexin is important for lens growth and transparency. Diverse gap junction channels formed by α3 and α8 subunits are important for the differentiation, elongation and maturation of lens fiber cells. Aberrant gap junction communication, caused by alterations of channel assembly, channel gating or channel conductance, can lead to different types of cataracts. These findings provide some molecular insights for essential roles of connexins and gap junctions in lens formation and the establishment and maintenance of lifelong lens transparency.     

C-Phycocyanin Modulates Selenite-Induced Cataractogenesis in Rats

The present investigation is aimed to evaluate the anticataractogenic potential of C-phycocyanin (C-PC), extracted and purified from Spirulina platensis. Enucleated rat lenses were maintained in vitro in Dulbecco’s modified Eagle medium (DMEM). Group I contained DMEM, Group II and Group III contained 100 μM of sodium selenite, Group III was subdivided into three viz IIIa, IIIb, IIIc supplemented with 100, 150, 200 μg of C-PC respectively. In the in vivo study, on tenth day post partum: Group I rat pups received an intraperitoneal injection of saline, Group II, IIIa, IIIb, and IIIc rat pups received a subcutaneous injection of sodium selenite (19 μmol/kg bodyweight) Group IIIa, IIIb, IIIc also received an intraperitoneal injection of 100, 150, 200 mg/kg body weight of C-PC, respectively, from postpartum days?9–14. On termination of the experiment, the lenses from both in vitro and in vivo studies were subjected to morphological examination and subsequently processed to estimate the activities of antioxidant enzymes namely superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, levels of reduced glutathione and lipid peroxidation products. Sodium selenite-exposed, C-PC-treated rat lenses (Group IIIc), showed significant restoration of antioxidant enzyme activity (p?<?0.05) when compared to their counterpart Group II. Group IIIc conserved the levels of GSH and lipid peroxidation products at near to normal levels as compared with Group II. Results conclude the possible role of C-PC in modulating the antioxidant enzyme status, thereby retarding sodium selenite-induced cataract incidence both in vitro and in vivo.     

Age-Related Changes of Elements in the Anterior Commissures and the Relationships Among their Elements

To elucidate compositional changes of the anterior commissure with aging, the authors investigated age-related changes of elements in the anterior commissures and the relationships among their elements. After ordinary dissection at Nara Medical University was finished, the anterior commissures were resected from 45 subjects, ranging in age from 70 to 101 years. The subjects consisted of 22 men and 23 women. After ashing with nitric acid and perchloric acid, the element content of the anterior commissures was determined by inductively coupled plasma-atomic emission spectrometry. The seven element contents of Ca, P, S, Mg, Zn, Fe, and Na did not change significantly in the anterior commissures with aging. Regarding the relationships among their element contents, significant correlations were found among the contents of Ca, Mg, Zn, and Na in the anterior commissures. The gender difference that the Zn content was significantly higher in men than in women was found in the anterior commissure.     

An assessment of the calcium content of rat liver mitochondria in vivo.

The effect of systematically altering the isolation conditions on the total calcium content of mitochondria isolated from perfused rat liver was examined. We showed that, under most isolation conditions, significant redistributions of mitochondrial calcium occurred resulting in up to 5-fold changes of the total calcium content. Mitochondrial Ca2+ flux inhibitors such as Ruthenium Red and nupercaine were only partially effective in inhibiting such redistributions. We present evidence indicating that the total calcium content of rat liver mitochondria in situ may approximate 2 nmol X (mg of protein)-1.     

Liver asialoglycoprotein receptor levels correlate with severity of alcoholic liver damage in rats.

It has been demonstrated that the oral administration of ethanol (Lieber-DeCarli liquid diet) to rats results in a decreased expression and content of the asialoglycoprotein receptor (ASGP-R) in the resultant fatty liver. In the present study, we wanted to determine whether the extent of impaired receptor content was correlated with the severity of liver pathology by using the intragastric feeding model. When ASGP-R protein and mRNA levels were measured in animals infused with ethanol or dextrose in the presence of fish oil (FO) or medium-chain triglyceride as the source of fat, more significant impairments to the ASGP-R were observed in the FO-ethanol group compared with the medium-chain triglyceride-ethanol group. Furthermore, only the FO-ethanol group showed pathological liver changes. These results demonstrate that a correlation exists between the progression of alcohol-associated liver injury, as defined by the severity of liver pathology, and an ethanol-induced decline in ASGP-R content.