Synthetic Activity Of Cells Research Articles

Immune Status of Turkeys in Industrial Cultivation Conditions

The intensification of turkey farming is accompanied by the impact of various dangerous factors on the poultry’s body. This leads to a violation of metabolic processes, an imbalance in the absorption of nutrients, and immunity decrease in turkeys, which determines the relevance of the study in this area. In this regard, figuring out when immunity reduces and, in the future, increasing its effectiveness in the body of turkeys, improving metabolic processes, growth, and development of poultry determined the purpose of this study. It was found that during the first six weeks of intensive cultivation of turkeys, the body’s immune response, the activity of non-specific resistance factors on the influence of biological factors and the intensification of poultry feeding processes were effective. However, from 42 days of poultry rearing, depletion of the immune defence of the turkey body and a decrease in the activity of non-specific resistance factors were observed, which is manifested by a likely decrease in the content of total protein, albumins, and class G and M immunoglobulins in the blood of poultry (P<0.01). Analysis of the protein spectrum of turkey blood serum indicates an immunosuppressive state of the body, probably caused by the action of biological factors. By 42 days, the bactericidal and lysozyme activity of blood serum, phagocytic activity of neutrophils, and T-cell activity of lymphocytes decrease in turkeys. Nonspecific changes in the components of the functional element of the liver were detected, which are accompanied by an increase in the number of lipid inclusions of various sizes, destructive changes in mitochondria, and a decrease in the synthetic activity of cells. Studies of the immune state of the body of turkeys, the activity of factors of non-specific resistance and liver in the cycle of intensive cultivation from 7 to 120 days revealed the presence of a critical period of decrease in the effectiveness of the body’s protection and further adequate correction. The results obtained during this study will be used for educational purposes, implemented in growing turkeys in the production conditions of “Indychka” LLC (Sumy Oblast). They are offered to all farms in Ukraine to produce turkey farming products

Immunophenotypic Characteristics of Macrophages in Liver and Peritoneal Area of Animals with Model of Diabetes Mellitus Type 1 and Their Corrections with Sodium Aminodihydrophthalazindione In Vitro

The goal of this study was to investigate the morphological and functional characteristics of macrophages of various localizations isolated from intact animals and animals with the model of diabetes mellitus type 1 (DM1) stimulated for 24 and 72 h in culture with sodium aminodihydrophthalazinedione (ADPN), a macrophage modulator drug. The experiments have been performed on cultures of rat macrophages obtained from the liver and peritoneal cavity. The following morphometric parameters were determined: the area of the cell, cytoplasm and nucleus, as well as the nuclear–cytoplasmic ratio (NCR). The phenotype of macrophages was determined by the expression of CD163 (macrophages M2) and CD80 (macrophages M1) markers. The functional activity of macrophages was assessed by the level of cytokines IL-1α, IL-10, and TNFα. It was found that ADPN altered the morphometric parameters (the size of cells and nuclei decreased, while NCR increased) and synthetic activity of cells (increased IL-10 in macrophages of the peritoneal cavity and IL-1α and TNFα in liver macrophages) during the first 24 h of cultivation. After 72 h, ADPN caused a decrease in the levels of IL-10 and TNFα and an increase in the level of IL-1α in all studied cell populations. ADPN did not affect the expression level of macrophage markers M1 and M2.

Histological and ultrastructural changes in turkey liver under intensive rear-ing and influence of xenobiotics

Liver problems of various etiologies in turkeys have been reported in many countries for the last 20 years. Poultry dies having no clinical manifestations of the disease, and at pathological autopsy, diffuse haemorrhages and marked dystrophic changes of the organ are noted. To date, there are several factors that can cause such changes, these are unbalanced amino-acid feed, insufficient calcium, biotin, selenium, the very high energy content of feed; zootechnical factors – limited mobility of birds due to cage density violations, high temperature; genetic factors – the influence of estrogens; infectious factors – E. coli, Clostridium, and viruses of Picornaviridae family. The article describes the histopathological and ultrastructural changes in the turkey liver under the influence of various factors. The material for the research was obtained from a farm where turkeys of the “Hybrid Converter” cross are grown, same age, fed with a standard diet that changed according to the technological map of cultivation. On the 50th day of life, a pathological autopsy of the dead poultry was performed, pieces of liver were selected for histological and ultrastructural examination. The visual assessment revealed significantly enlarged liver, the colour from dark red to light brown, flabby consistency. In some cases, diffuse fatty infiltrations of hepatocytes were histologically revealed, in other cases, focal necrosis with the growth of the connective tissue and the formation of massive perivascular couplings were registered. Large vacuolar fatty degeneration of hepatocytes with subsequent development of fibrosis indicates chronic intoxication, probably caused by slow breakdown of fatty acids in cells due to insufficient oxidative phosphorylation, as well as reduced levels of lipotropic factors: choline, methionine and the vitamins. At the ultrastructural level, a large number of lipid inclusions of various sizes, dystrophic changes in mitochondria were observed, which indicates a decrease in the synthetic activity of cells.

Ageing does not result in a decline in cell synthetic activity in an injury prone tendon.

Advancing age is a well-known risk factor for tendon disease. Energy-storing tendons [e.g., human Achilles, equine superficial digital flexor tendon (SDFT)] are particularly vulnerable and it is thought that injury occurs following an accumulation of micro-damage in the extracellular matrix (ECM). Several authors suggest that age-related micro-damage accumulates due to a failure of the aging cell population to maintain the ECM or an imbalance between anabolic and catabolic pathways. We hypothesized that ageing results in a decreased ability of tendon cells to synthesize matrix components and matrix-degrading enzymes, resulting in a reduced turnover of the ECM and a decreased ability to repair micro-damage. The SDFT was collected from horses aged 3-30 years with no signs of tendon injury. Cell synthetic and degradative ability was assessed at the mRNA and protein levels. Telomere length was measured as an additional marker of cell ageing. There was no decrease in cellularity or relative telomere length with increasing age, and no decline in mRNA or protein levels for matrix proteins or degradative enzymes. The results suggest that the mechanism for age-related tendon deterioration is not due to reduced cellularity or a loss of synthetic functionality and that alternative mechanisms should be considered.

Age-Related Features of Cardiomyocyte Ploidy in Patients with Hypertrophic Obstructive Cardiomyopathy.

Endomyocardial biopsy samples of the interventricular septum from patients with hypertrophic cardiomyopathy isolated during myectomy were examined. We revealed significant variability of cardiomyocyte ploidy (from 2.9c to 13.5c) that directly correlated with myocyte size; the maximum ploidy was found in young patients. Ultrastructural signs of increased contractile and synthetic activity of cells were observed in patients with high DNA content in cardiomyocytes.

Use of milk epithelial cells to study regulation of cell activity and apoptosis during once-daily milking in goats

Generally, once-daily milking (ODM) decreases milk yield. This effect may be the consequence of a decrease in mammary epithelial cell (MEC) activity or a reduction in their number. The aim of this study was to determine the effect of ODM on the synthetic activity and rate of apoptosis of MEC using a non-invasive method. Eight Alpine goats were subjected to ODM or twice-daily milking for two 5-week periods. MECs were purified by centrifugation and immunocytochemical binding in milk after 1 and 5 weeks of each period. mRNA levels of some proteins involved in lactose and milk protein synthesis and in apoptosis were evaluated using real-time PCR. Isolation of MEC from milk was a useful method to investigate transcriptional regulation in a timeline study. ODM induced greater decreases in milk, lactose and protein yields after 1 week than after 5 weeks. This suggests an adaptation of the mammary gland to ODM, which reduces the inhibitory effect of this practice. Reductions in milk component yields were associated with lower α-lactalbumin transcripts, suggesting a transcriptional decrease of lactose synthesis during ODM. Glucose transporter GLUT1 transcripts were downregulated under ODM, suggesting that lactose precursor uptake by MEC might be involved in the regulation of lactose synthesis. κ-Casein mRNA levels tended to be lower during ODM. ODM increased levels of the pro-apoptotic transcript Bax after both 1 and 5 weeks, but no variation was observed in the Bax/Bcl-2 ratio. ODM affected cell synthetic activity through transcriptional regulation and may have induced apoptosis. The reduction of the negative effect of ODM on milk yield suggests that Alpine goats are able to adapt to ODM. Further studies are needed to investigate the effect of ODM on MEC turnover.

Effects of Radiation Therapy on Chondrocytes In Vitro

The negative irradiation complications of growth loss leading to limb length asymmetry and pathological fracture incurred following radiation therapy in pediatric patients has led to a renewed interest in understanding the specific effects of irradiation on the growth plate and the surrounding bone. In the present report, we examined the radiation therapy effects on primary rat growth cartilage chondrocytes in order to determine the chondrocyte radiosensitivity relative to other bone cell constituents and tumor cells, the postirradiation temporal progression of radiation-induced alterations in chondrocyte function, and the time course for the functional restoration of chondrocyte pathways that drive the eventual recovery in growth function. We employed an in vitro primary rat costochondral growth cartilage cell culture model system to evaluate the radiation therapy effects on proliferative chondrocytes using serial radiation doses (0-20 Gy) that are well within the clinically relevant range. Following irradiation, all of the following occurred in a dose-dependent manner: proliferation decreased, cytotoxicity increased, several markers of apoptosis increased, markers of radiation-induced cellular differentiation increased, and cell synthetic activity was disturbed. Alterations in proliferation, cell death, and induction of apoptosis are likely due to a transient radiation-induced derangement of the parathyroid hormone-related protein-Indian hedgehog proliferation-maturation pathway. Alterations in cellular differentiation and cell synthetic activity are novel observations for chondrocytes. Further, these results correspond very well to our previous work in an in vivo Sprague-Dawley rat model, making this model particularly relevant to researching the radiation therapy effects on longitudinal growth.

Surgical removal of articular cartilage leads to loss of chondrocytes from cartilage bordering the wound edge.

A number of arthroscopic procedures that are used in the treatment of focal cartilage lesions or osteoarthritic joints, such as shaving, débridement, and laser abrasion, involve the removal of both diseased and healthy articular cartilage. The excision of such tissue has the effect of generating lesions within the articular cartilage. The fate of the chondrocytes that border such lesions has not been evaluated. The purpose of this investigation was to ascertain whether the surgical creation of lesions in articular cartilage induces irreversible loss of chondrocytes over time from tissue bordering the wound edge and to determine whether the synthetic activity of cells in this region is compromised. Partial-thickness defects of defined dimensions were created in the femoral condyle and/or trochlear groove of rabbits and miniature pigs. Cell volumes, cell volume densities, and numerical cell densities within tissue close to (within 100 micro m) and remote from (control site) the wound edge were determined by quantitative histomorphometry at various time intervals up to six months after surgery. Rates of proteoglycan synthesis by cells in both regions were determined by quantitative autoradiography following (35) S-sulphate labeling in vivo. The surgical creation of partial-thickness lesions in articular cartilage induced a significant and long-term loss of cells from tissue near the wound edge. However, the surviving cell population maintained a normal rate of matrix proteoglycan deposition. This study illustrates that maintenance and remodeling of cartilage matrix close to wound edges in articular cartilage lesions is compromised, since fewer cells, with an unchanged metabolic activity rate, are left to sustain matrix domains.

Effect of estradiol dipropionate on protein synthesis in oocytes and ovary of the sea urchin Strongylocentrotus intermedius at different stages of the reproductive cycle

We studied protein synthesis in the oocytes and ovary of the sea urchin Strongylocentrotus intermedius at different stages of the reproductive cycle after treatment with estradiol dipropionate. During the early development of oocytes and active gametogenesis, this estrogen induced the incorporation of 3H-leucine in the oocytes. The changes in synthetic activity of cells were accompanied by an elevated efficient incorporation of free amino acid in proteins due to its increased pool, increased tissue permeability for precursors, and increased rate of protein synthesis. Before spawning, estradiol dipropionate did not cause any changes in protein synthesis. After estradiol dipropionate treatment, the inhibitors of protein synthesis, puromycin and actinomycin D, decreased the intensity of 3H-leucine incorporation in the oocytes and protein synthesis in the ovary. The involvement of estradiol dipropionate in the regulation of protein synthesis in the sea urchin gonad is discussed.

The Effect of Estradiol Dipropionate on the Rate of Protein Synthesis in the Testicle of the Sea Urchin Strongylocentrotus nudus

The effect of estradiol dipropionate on the rate of protein synthesis in the testicle of the mature sea urchin Strongylocentrotus nuduswas studied. The injection of this estrogen considerably intensified 3H-leucine incorporation into the gonad. The change in the level of cell synthetic activity is assumed to be associated with a rise in effective incorporation of 3H-leucine into proteins following an increase in its intracellular level, and with an increase in the protein synthesis rate. The participation of sex hormones in the regulation of protein synthesis in the S. nudusgonad is discussed.

In vitro characterization of mesenchymal stem cell-seeded collagen scaffolds for tendon repair: effects of initial seeding density on contraction kinetics.

Mesenchymal stem cells (MSCs) were isolated from bone marrow, culture-expanded, and then seeded at 1, 4, and 8 million cells/mL onto collagen gel constructs designed to augment tendon repair in vivo. To investigate the effects of seeding density on the contraction kinetics and cellular morphology, the contraction of the cell/collagen constructs was monitored over time up to 72 h in culture conditions. Constructs seeded at 4 and 8 million cells/mL showed no significant differences in their gross appearance and dimensions throughout the contraction process. By contrast, constructs seeded at 1 million cells/mL initially contracted more slowly and their diameters at 72 h were 62 to 73% larger than those seeded at higher densities. During contraction, MSCs reoriented and elongated significantly with time. Implants prepared at higher seeding densities showed more well aligned and elongated cell nuclei after 72 h of contraction. Changes in nuclear morphology of the MSCs in response to physical constraints provided by the contracted collagen fibrils may trigger differentiation pathways toward the fibroblastic lineage and influence the cell synthetic activity. Controlling the contraction and organization of the cells and matrix will be critical for successfully creating tissue engineered grafts.

Cell synthesis, proliferation and apoptosis in human dental periapical lesions analysed by in situ hybridisation and immunohistochemistry.

The role of structural and host defensive cells in periapical lesions has been assessed previously by morphometric and immunohistochemical studies. The aim of this study was to investigate the function of peri- apical cells by employing molecular techniques to estimate the cell synthetic activity, proliferation and apoptosis in these lesions. We specifically sought answers to the following questions. Which cells of the periapical lesions are quiescent or actively synthesising proteins? Do immune cells proliferate in this region in the same way as epithelial cells proliferate? Furthermore do cells in peri- apical lesions undergo apoptosis, and if so which cells exhibit this programmed cell death? Twenty-five periapical tissue samples (15 granulomas and 10 radicular cysts) were assessed. Poly-adenosine (poly (A)) RNA and ribosomal RNA (rRNA) bearing cells in formalin-fixed/paraffin-embedded peri- apical tissues were analysed by in situ hybridization (ISH) using digoxigenin-labelled oligo d (T) and 28S rRNA probes respectively in order to estimate cell synthetic activity. Furthermore, S-phase proliferating and cycling cells were examined by ISH using a histone probe and Ki-67 immunostaining so as to assess cellular proliferation. Mononuclear cells were further differentiated by immunohistochemistry (IHC) as T cells, B cells and macrophages. Apoptotic cells were determined by in situ end-labelling methodology for detecting fragmented DNA. Poly (A) RNA (mostly messenger RNA) and 28S rRNA-expressing cells were detected in all samples. Plasma cells exhibited strongest staining for the two probes, with slight to moderate staining found in the epithelium, fibroblasts, macrophages, endothelial cells and lymphocytes, whereas almost all polymorphonuclear leucocytes (PMN) were negative for these probes. A few histone mRNA-expressing cells were detected in basal and suprabasal epithelial cells and mononuclear cells in 15/25 cases but their reactivity was weak. Ki-67 positive cells were found in all samples and their numbers were generally higher than histone mRNA positive cells. Apo- ptotic cells were detected in 23/25 cases and the majority of apoptotic cells were PMN which were engulfed by large cytophagocytic macrophages. This study indicates that in dental periapical lesions, apoptosis occurs predominantly in PMN. It is evident that most cells apart from PMN are exhibiting synthetic activity but only epithelial cells undergo proliferation which implies that immune cells must proliferate at distant lymph nodes and travel to the periapical lesion rather than proliferating within the lesion. These results suggest considerable advantages in estimating gene expression within cells in addition to the immunohistochemical detection of cells to determine cell activity at inflamed sites. Clearly, functional cell synthetic activity, resolution and clearance systems operate in peri- apical cystic and granuloma lesions.

Effects of angiotensin converting enzyme inhibition on glomerular number, juxtaglomerular cell activity and renin content in experimental unilateral hydronephrosis

To determine the effects of hydronephrosis on glomerular number and juxtaglomerular cell synthetic activity and the protective influence of angiotensin converting enzyme inhibition. A comparison of sham and contralateral kidneys with 8-week ipsilateral ureteral ligated hydronephrotic kidneys in BALB/c mice. Enalapril was administered from 5 weeks in additional sham and hydronephrotic kidney groups. Renin and prorenin immunohistochemistry was applied to sections of perfusion-fixed kidneys at the light and electron microscope level. Glomerular number was estimated by a physical disector-fractionator stereological method. An enzyme kinetic renin assay was performed in kidney tissue and plasma. Glomerular number in hydronephrotic kidneys decreased significantly compared with sham and contralateral kidneys. Renin content in hydronephrotic kidneys did not change compared with sham or contralateral kidneys, but the renin content in the glomerulus was significantly greater in hydronephrotic than in contralateral kidneys and similar to in sham kidneys. Contralateral kidneys enlarged significantly and their total renin content decreased significantly compared with hydronephrotic and sham kidneys. Plasma renin was unchanged. Fewer juxtaglomerular cells were labelled for renin and prorenin in contralateral than in hydronephrotic or sham kidneys. Granulopoiesis and exocytotic profiles were markedly greater in hydronephrotic than in contralateral or sham kidneys. Following enalapril, glomerular number was significantly higher in hydronephrotic kidneys and renin content increased proportionally more in contralateral than in hydronephrotic or sham kidneys. Hydronephrosis for 8 weeks results in atrophy of 50% of glomeruli and exerts an inhibitory influence on contralateral juxtaglomerular cells while augmenting ipsilateral renin production per remaining glomerulus with maintenance of plasma renin. Enalapril preserves glomeruli and reverses the contralateral inhibitory influence, suggesting an angiotensin-related mechanism.

4-MAPC, a 5 alpha-reductase inhibitor, reduces rat ventral prostate weight, DNA, and prostatein concentrations.

Several compounds, such as 4-MAPC (4-methyl-3-oxo-4-aza-5 alpha-pregnane-20- carboxylate), that inhibit conversion of testosterone (T) to dihydrotestosterone (DHT) by 5 alpha-reductase have been demonstrated to reduce prostate size in rats and dogs. The current studies were undertaken to determine if this effect is due to a reduction in cell number, in epithelial cell synthetic activity, or both. Eight-week-old intact rats were treated daily for 14 days with sesame seed oil, 4-MAPC (10 mg/kg), 4-MAPC + testosterone propionate (TP, 1 mg/kg), or 4-MAPC + TP (3 mg/kg). Rats were killed 24 hours after the last injection. In the animals treated only with 4-MAPC, ventral prostate weight was reduced 37%, but the 14% reduction in total DNA was not significant. The mean intraprostatic concentration of prostatein, a major secretory protein, was reduced 45% (P less than 0.05). The 3 mg/kg dose of TP increased ventral prostate weight, prostatein concentrations, and acid phosphatase activity, even though DNA/ventral prostate was similar to that in control animals. These observations indicate that the reduction in ventral prostate weight in adult rats is due in part to a reduction in cell number, but the primary effect was due to a reduction in synthetic activity, and possibly atrophy of the epithelial cells. Furthermore, TP in pharmacologic doses increased ventral prostate weight and synthetic activity without increasing DNA.

Extracellular matrix molecules and their receptors: an overview with special emphasis on periodontal tissues.

Knowledge of extracellular matrix molecules and their cell receptors has increased exponentially during the last 2 decades. It is now known that the structure and function of each tissue is based on specific combinations of matrix molecules. The major constituents of the extracellular matrix are collagens, proteoglycans, and adhesive glycoproteins. The rapid development of biochemical, molecular biological, and immunological research has revealed a lot of interesting details pertaining to these molecules. Several new collagen types have been discovered. In addition to being responsible for the strength and form of tissues, each collagen type has specific sequences providing them with special features such as flexibility and the ability to interact with other matrix molecules and cells. Proteoglycans are another large group of matrix molecules with a variety of functions. Proteoglycans play an important role in tissue resilience and filtering. Some proteoglycans have a capacity to specifically bind other matrix molecules and growth factors, while others act as matrix receptors on the cell surface. An important part of regulation of the cell behavior is played by adhesive glycoproteins belonging to the fibronectin and laminin families. Several isoforms of fibronectin and laminin that result from alternative RNA splicing serve specific functions such as controlling the attachment, migration, and synthetic activity of cells. A major group of cell receptors for cell-matrix and cell-cell interactions is termed integrins. The integrins are cell surface proteins composed of two polypeptides whose structure dictates the specificity of each receptor. The cytoplasmic domain of the integrins interacts with cytoskeletal elements within the cell, and thereby relays the information from the extracellular space into the protein synthesis machinery. The expression of the integrins is controlled by the extracellular matrix and growth factors, most notably TGF beta. During periodontal diseases several aspects of the cell-matrix interactions may be disturbed. Therefore, an understanding of the special features of the extracellular matrix and their receptors in periodontal tissues is a prerequisite for developing new approaches to the prevention and treatment of periodontal diseases.

Activin A/erythroid differentiation factor induces thromboxane A2 synthetic activity in murine erythroleukemia cells

Activin A, a protein homologous to transforming growth factor beta, was shown to induce hemoglobin synthesis in murine erythroleukemia (MEL) cells and was also termed erythroid differentiation factor (EDF) (Eto, Y., Tsuji, T., Takezawa, M., Takano, S., Yokogawa, Y., and Shibai, H. (1987) Biochem. Biophys. Res. Commun. 142, 1095-1103). We found that activin A/EDF also induced thromboxane (TX) A2 synthetic activity in these cells. Synthesis of TXA2 from arachidonic acid is catalyzed by cyclooxygenase and TX synthase. Activin A/EDF induced the latter TX synthase activity, whereas the cyclooxygenase activity was constitutively expressed. The induction of this enzyme activity was inhibited by cycloheximide, suggesting that activin A/EDF induced de novo protein synthesis of TX synthase. Furthermore, we studied the relationship between the induction of TXA2 synthetic activity and erythroid differentiation in MEL cells, since the former is not an erythroid phenotype. We found 1) that the two responses to activin A/EDF were distinctly affected by the initial cell density; 2) that the dose-response curves for activin A/EDF were similar (ED50 = approximately 100 pM), whereas the time course of induction of TXA2 synthetic activity was much faster; and 3) that other erythroid differentiation inducers of MEL cells, namely dimethyl sulfoxide and hexamethylene bisacetamide, had little or no effect on TXA2 synthesis. These results indicate that activin A/EDF induces TXA2 synthetic activity independently of erythroid differentiation.

Proliferative and synthetic activity of cells from reoccluded distal anastomosis and pharmacological therapy of this postoperative complication

Proliferative and synthetic activity of cells from reoccluded distal anastomosis and pharmacological therapy of this postoperative complication

Oxidant-induced Cytotoxicity: A Challenge for Metabolic Modulation

Oxidant injury to cells and tissues, induced by reactive oxygen metabolites such as superoxide anion, hydrogen peroxide, and hydroxyl radical, involves chemical alterations in proteins, lipids, carbohydrates, and nucleic acids (1-3). Perturbations to the first three of these molecules may impair membrane and other cellular activities; however, such damage can usually be repaired by new synthesis, and damaged tissues can be replaced by cell replication. However, damage to nucleic acids in general, and DNA in particular, interferes with the cell's synthetic activity and more importantly with its ability to replicate and replace damaged cells. In addition to the replicative impairment that may result from oxidant injury, reactive oxygen metabolites can induce sufficient DNA damage to activate metabolic pathways that lead to rapid cell lysis and death (3). This more fulminant type of cell death is mediated by activation of the chromosomal enzyme, poly(ADP-ribose) polymerase and consequent alterations in NAD metabolism (3-7). Figure I shows the central regulatory role played by NAD and the potential for drastic alterations in cellular metabolism consequent to variations in NAD levels. Most cells . maintain a relatively steady state level of NAD. They can take up nicotinamide and combine it with phosphoribosylpyrophosphate to form nicotinamide mononucleotide which can then be condensed with ATP to form NAD (8). NAD serves as cofactor for glycolysis and the tricarboxylic acid cycle which provides most of the ATP for energy-dependent processes. NAD is the precursor for NADP, which serves as cofactor for several important pathways including the pentose shunt, bioreductive synthetio pathways, and maintenance of reduced glutathione pools. The bottom of Figure 1 indicates that oxidants can produce direct DNA strand breaks and that they can also produce DNA damage that is processed into DNA strand breaks by cellular repair enzymes. DNA strand breaks activate the chromosomal enzyme poly(ADP-ribose) polymerase to utilize NAD for the production of poly(ADP-ribose), which facilitates the DNA repair process. Poly(ADP-ribose) polymerase shows negligible levels of activity until DNA is

Effect of cell donor age on the synthetic properties of fibroblasts obtained from phenytoin‐induced gingival hyperplasia

Diploid fibroblasts obtained from explants of human gingiva and maintained in vitro undergo a several-fold decrease in protein and collagen synthesis as a function of increasing donor age. Using drug-induced gingival hyperplasia as a model, we performed experiments to learn whether fibroblasts derived from hyperplastic tissue behave in a similar manner. Fibroblast strains were established from explants of hyperplastic gingiva obtained from 10 patients chronically ingesting phenytoin and ranging in age from 9 to 45 years. Protein production and degradation were compared to previously reported data similarly obtained from periodontally normal donors ranging in age from 12 to 68 yr. The total quantity of protein and collagen produced by the phenytoin cells was significantly greater than previously reported for cells from normal gingiva. No donor age-related decrease in protein and collagen production nor in the proportion of cell synthetic activity committed to collagen production was observed for cultures of phenytoin cells. The gross pattern of proteins produced, as assessed by 2-dimensional gel electrophoresis, was unrelated to donor age in both normal and phenytoin cells, but three polypeptides ranging in size from about 20 kD to 40 kD that were not found in the cultures of normal cells were produced by five of seven phenytoin cells strains. The observations demonstrate that the phenytoin cells do not undergo the donor age-dependent decrease in synthesis observed for normal cells. This abnormality may account in part for the phenytoin-induced hyperplasia. The phenytoin cells appear to be a unique phenotype.

Nucleolar organizer regions relate to growth fractions in human breast carcinoma

An argyrophil stain for nucleolar organizer regions (NORs) has recently been applied to paraffin sections of human tissues. This report describes a positive relationship between the mean numbers of AgNOR sites per nucleus and tumor growth fraction, as determined by immunostaining with the monoclonal antibody Ki-67, in 83 malignant breast tumors ( P < .01). This relationship supports recent suggestions that the NOR count may reflect cell synthetic activity and hence, proliferation. AgNOR counts correlated inversely with immunocytochemically assessed estrogen receptor content ( P < .002), but there was no relationship between the AgNOR count and primary tumor size, histologic grade, axillary node status, or patient age. A significant difference ( P < .00001) was found between the AgNOR counts in 64 benign breast lesions (mean, 2.05) and 85 malignant breast neoplasms (mean, 5.46). The limitations of the silver staining technique and the problems of reproducibility in AgNOR counting are detailed.