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S Segal

Publications and source records attributed to S Segal.

At least 379 records · Page 21Linked to original sources

L-Proline transport by isolated renal tubules from newborn and adult rats.

Proline uptake and metabolism has been examined in newborn Sprague-Dawley rat kidney and compared to that in adult animals. [14C]-Proline uptake by renal tubule fragments from newborn rats occurs at the same initial rate as in adult tubules, but at physiologic concentrations achieves significantly higher intracellular levels after 15 min of incubation. Considerable metabolism of the proline taken up was observed in tissue of both ages. Analysis of acid soluble and insoluble tubule fractions from newborn and adults indicates similar degrees of proline incorporation into protein and oxidation to CO2 relative to the amount of radioactivity taken up. A major difference exists, however, with respect to the labeled components of the acid extract: adult tubules convert [14C]-proline to metabolites at a rate twice that of newborn. Analysis of concentration-dependent uptake data reveals two distinct entry systems for proline in both isolated newborn and adult tubules. No difference in the Km or Vmax was found between the young and mature tubules.

Age Factors↗

Galactose metabolism in suckling and adult isolated rat hepatocytes.

The metabolism of galactose during the postnatal developmental period has been examined in isolated parenchymal cells of livers from fasted rats aged 7, 14, 21, 28, and 42 days, by measuring the disappearance of 1 and 4 mM 1-[14C]galactose from the incubation media as well as [14C]galactose conversion to [14C]glucose and [14C]lactate and oxidation to [14CO2]. Cells from 7- and 14-day-old suckling animals consistently utilized galactose more rapidly (4 times greater) than those of adult. Hepatocytes of suckling rats also converted greater amounts of galactose to glucose and oxidized galactose 2-3 times faster than the adult. The conversion to glucose by suckling cells occurred with minor recycling of labeled C-1 galactose to C-6 of glucose. A comparison with [1-14C]glucose as substrate showed that oxidative rates by the young cells were 3 times faster for galactose than for glucose. This was not due to the presence of a direct galactose oxidative pathway as assessed by the [14CO2] yield from C-1 and C-2 labeled galactose. Incubation of hepatocytes with galactose appeared to augment the production of glucose from endogenous precursors, 33 nmoles/mg cells in 30 min. The enhanced glucose output from endogenous sources in isolated suckling hepatocytes incubated with galactose contrasts with the sugar's suppression of glucose output observed in isolated perfused liver even though galactose metabolism in both preparations are similarly greater in the suckling than in the adult.

Animals↗

Lymphomatoid granulomatosis with impaired cellular immunity. Eight year survival without treatment.

Lymphomatoid granulomatosis (LYG), a non-neoplastic lymphoreticular disorder, was diagnosed in a 65-year-old woman. Chest radiographs demonstrated bilateral lower lobe nodular infiltrates. Percutaneous needle biopsy of the lung showed an infiltrate composed of plasma cells, lymphocytes and large histiocytic-like cells. Impairment of cellular immunity was found by in vivo as well as by in vitro tests. The clinical condition of the patient has remained stable for the last eight years without specific treatment.

Aged↗

Evidence for beta 2-microglobulin-like and H-2-like antigenic determinants in Drosophila.

Serologic evidence for the existence of beta 2-microglobulin-, (beta 2m) like and H-2-like antigenic determinants in Drosophila are presented. Drosophila-cultured cells and larvae extract were shown to react specifically with a rabbit anti-rat beta 2m and a rabbit anti-mouse beta 2m antisera. G-200 pooled fractions from Drosophila larvae were shown to react with beta 2m-eluted and glycin-absorbed antisera, but not with beta 2m-absorbed or glycin-eluted antisera. These fractions also quantitatively inhibited the heterologous reaction between the anti-beta 2m antisera and purified human and rat beta 2m. The lack of reactivity of other rabbit antisera or normal serum with Drosophila, as well as the efficiency of absorption of anti-beta 2m reactivity by either rat or KCO% (Drosophila) cells, further supports the presence of beta 2m epitopes on Drosophila cells. Data are also presented showing that certain anti-H-2 alloantisera react with Drosophila. That this reactivity is indeed due to anti-H-2 antibodies is suggested by several lines of evidence, including the removal of cytotoxic alloantibodies and lack of reactivity by other mouse antisera and monoclonal reagents. Preliminary data suggest that the H-2-like and beta 2m-like determinants are physically associated on the cell surface of Drosophila cells. These findings have important implications on current concepts concerning the evolutionary origin and physiologic role of beta 2m and the major histocompatibility complex.

Animals↗

Mixed lymphocyte reactivity nonresponsiveness in couples with multiple spontaneous abortions.

We have endeavored to ascertain the possible existence of deviant recognition of antigens controlled by the major histocompatibility complex by lymphocytes originating in couples with a history of multiple spontaneous abortions of unknown cause. Human leukocyte antigen (HLA) typing, as well as one-way mixed lymphocyte culture (MLC) tests, were performed on 33 couples with multiple spontaneous abortions and subsequently compared to 30 fertile couples with no abortion history. No significant differences in the frequency of HLA were detected between the fertile and infertile groups. The frequencies of shared HLA-A and -B antigens among members of the couples were similar in both groups. In 70% of the cases studied, the husbands revealed a depressed cellular response to their respective wives; whereas reactivity to cells from other women proved normal. However, the response of allogeneic controls to these infertile women was normal. This implies that women with multiple abortions are not poor stimulators. The specific one-way decrease in the MLC reactivity appeared to be mediated by female-derived suppressor mechanisms and may be considered as one of the causes of the interrupted development of pregnancy in vivo.

Abortion, Habitual↗

Loss of expression of transplantation antigens encoded by the H-2K locus on Lewis lung carcinoma cells and its relevance to the tumor's metastatic properties.

The Lewis lung carcinoma 3LL, a C57BL/6 (H-2b)-originated spontaneous tumor that progresses at the site of transplantation in all mouse strains, produced lung metastases only in mice that shared the H-2D region and the non-H-2 genetic background with the tumor's strain of origin. In vitro cytotoxicity assays revealed that 3LL tumor cells were sensitive to lysis exerted by anti-H-2Db immune effector cells but were relatively resistant to lysis by anti-H-2Kb effector cells. In addition, the 3LL tumor cells could inhibit the anti-H-2Dd response against EL 4 (H-2b) target cells but had almost no inhibitory effect on the anti-H-2Kb response against the same targets. Immunization of C3HeB (H-2k) and B10.D2 (H-2d) mice with 3LL tumor cells resulted in antisera that could preferentially react with H-2Db-positive cells but reacted very weakly with H-2Kb-positive cells. The results indicate that 3LL tumor cells expressed H-2Db cell surface antigens at a high density and seemed to lack membrane glycoproteins that are encoded by the H-2Kb region.

Animals↗

Alterations in major histocompatibility complex phenotypes of mouse cloned T10 sarcoma cells: association with shifts from nonmetastatic to metastatic cells.

The T10 sarcoma, induced in a (C57BL/6J X C3HeB/-FeJ)F1 (H-2b X H-2k) mouse, grows locally (L-T10) and generates spontaneous lung metastases (M-T10). L-T10 cells were found to express the H-2b haplotype, whereas M-T10 expressed both the H-2b and H-2k haplotypes. Most L-T10 cloned cells expressed the H-2b haplotype and were not metastatic. The minority expressed both H-2k and H-2b and were metastatic. Serial transplantations of H-2k-negative clones always ended in spontaneous expression of the H-2k haplotype concomitantly with the acquisition of metastatic potency. The expressed H-2k seemed to be associated with the metastatic properties inasmuch as an H-2b-positive--H-2k-negative clone, which had lost the expressed H-2b and was temporarily H-2 negative, remained nonmetastatic until reexpression of the two haplotypes occurred. Serial transfers of H-2k-positive clones resulted in the maintenance of the expressed H-2k haplotype and the retention of metastatic capacity. A shift toward increased metastatic capacity correlated with H-2k expression occurred during serial transfers of every clone tested. Expression of major histocompatibility complex components, rather than their loss, may potentiate the metastatic capacity of tumor cells.

Animals↗

Developmental changes of glycine transport in the dog.

The renal clearance of amino acids was measured in canine pups between 5 days and 12 weeks of age. The reabsorption of glycine was incomplete at 5 and 21 days, indicating a physiologic aminoaciduria of immaturity. An adult pattern of 97-100% reabsorption appeared by 8 weeks of age. The uptake of glycine by isolated renal tubules from 5-day-old, 3-month-old and adult dogs was examined towards an understanding of the events underlying this aminoaciduria. The initial uptake of 0.042 mM glycine by isolated tubules from the newborn was lower than that of the adult, but after 30 min of incubation the newborn surpassed the adult. A steady state of uptake was not achieved by the newborn even after 90 min of incubation, while it was achieved in the adult after 30 min. The uptake by the 3-month-old tubules resembled the adult at the early time points and the newborn at later points. With 1.032 mM glycine, a similar relationship of uptake between adult and newborn tubules was found, except with this concentration, the uptake by both the newborn and adult tubules reached a steady state. The concentration dependence of glycine uptake showed two saturable transport systems with similar apparent Km and Vmax values after 30 min of incubation for all three age groups. Determination of glycine flux by compartmental analysis revealed decreased influx and efflux in the newborn, but with a greater decrease in efflux, compared to adult. These changes of influx and efflux which accompany renal tubule maturation could contribute to the increased intracellular amino acid levels and decreased reabsorption of amino acids seen in the immature dog.

Aging↗

The effect of azaserine upon the proline and methyl alpha-D-glucoside transport systems of rat renal brush-border membranes.

An inhibitory effect of azaserine on Na+ dependent proline and methyl alpha-D-glucoside transport of the rat renal brush-border membrane vesicles has been demonstrated. The inhibitory effects of azaserine were not the results of the drug disrupting the membrane vesicles as shown in osmolarity studies, nor did it affect the transport systems' affinities for Na+. Azaserine acts as a non-competitive inhibitor for the proline transport system in renal brush-border membranes by lowering 37% and 27% in the Vmax1 and Vmax2, respectively, when compared to that of control proline transport system. Azaserine had no effect upon the two Km values for proline uptake. Azaserine inhibition of methyl alpha-D-glucoside uptake by vesicles in the presence of 7.2 mM azaserine at 22 degrees C resulted in 66% increase in Km1 value and 44% decrease in Vmax1 as compared to that of control vesicles. There was no detectable effect upon the Km2 and Vmax2 of the methyl alpha-D-glucoside transport system. No effect of the drug was observed when sodium was equilibrated across the membrane, indicating that azaserine altered the driving force exerted by a sodium gradient. Azaserine only slightly affected the relative contribution of the two Km systems to total proline uptake. Contrary to the observed effect of azaserine upon the proline transport system, azaserine exerted a distinct effect upon the relative contribution to total uptake by the two Km systems in the low methyl alpha-D-glucoside concentration range. In the presence of 7.2 mM azaserine, the low-affinity, high-Km transport system becomes the major contributor to total methyl alpha-D-glucoside uptake by isolated renal brush-border vesicles.

Animals↗

Developmental pattern of cystine transport in isolated rat renal tubules.

Isolated renal cortical tubule fragments from rats ranging in age from less than 48 h to 15 weeks were used to examine the pattern of cystine uptake with development. Immature tubules took up cystine with a faster initial rate than mature tubules and did not reach a steady state by 60 min. By eight weeks of age, the timed uptake of cystine began to approach a steady state and between 8 and 11 weeks the uptake pattern achieved its adult form of reaching a steady state by 30 min of incubation. Analysis of the intracellular metabolism of the cystine taken up by the newborn tubules revealed that the majority had been reduced to cysteine with the formation of small amounts of reduced glutathione. Cystine entered the renal cortical tubule cell from the newborn via two saturable transport systems similar to the mature animal. The kinetic parameters of initial uptake of these two transport systems were similar in the mature and newborn animal except for a higher maximum transport velocity for the low Km, low capacity system in the newborn. Lysine inhibited cystine uptake by newborn tubules and this inhibition appeared to occur on the low Km, low capacity transport system similar to the adult. Cystine uptake was sodium dependent with an apparent affinity for sodium of 36 mequiv./l. From this data, the physiologic cystinuria of the immature animal does not appear to be refeable to a lower rate of influx as previously observed with the cortical slice. Other mechanisms should be sought to explain this phenomenon of immaturity.

Aging↗

The effect on amino acid transport of trypsin treatment of rat renal brush border membranes.

Trypsin treatment of isolated rat renal brush border membrane vesicles which preferentially releases L-leucine aminopeptidase (EC 3.4.11.2) decreases their ability to take up a variety of amino acids under Na+ -gradient conditions. Such treatment did not alter the osmotic properties of the vesicles nor affect their fragility. A linear correlation could be demonstrated between the L-leucine aminopeptidase activity of the membranes and the initial rate of uptake of L-leucine and L-proline. Velocity of uptake-concentration dependence studies with these substrates indicate that the major effect of trypsinization is to decrease the maximum velocity (Vmax1) of the low-Km high-affinity system with little effect on the Vmax2 of the high-Km low-affinity transport process and no effect on the apparent Michaelis constants of either. Although the data indicate that L-leucine aminopeptidase activity and uptake of l-leucine and L-proline are affected in parallel, they should not be construed to imply a role of the enzyme in the transport process, especially in view of the global decrease in the uptake of various amino acids and sugars.

Alkaline Phosphatase↗

RNA synthesis in cultured human placenta.

The in vitro synthesis of RNA in the human placental tissue, incubated in organ culture, was investigated. We followed the synthesis of the poly A(-) and poly A(+) RNA fractions, and investigated the distribution of the newly synthesized RNA among the subcellular fractions isolated from first and third trimester placentas. The poly A(-) RNA was the major fraction of the RNA synthesized in vitro. The incorporation of [3H]uridine into the poly A(+) RNA fraction was very low. As protein synthesis occurred during the entire incubation period, we suggest the presence of a pool of mRNA molecules in the form of mRNP particles.

DNA↗

Anti-tumor immunity in B-lymphocyte-deprived mice. III. Immunity to primary Moloney sarcoma virus-induced tumors.

Tumor induction and immunity to tumors were studied following the injection of Moloney sarcoma virus (MSV) into mice whose B-lymphocyte functions had been suppressed by the chronic administration of anti-IgM antibodies. Two preparations of MSV were used; one which gives rise to tumors which uniformly regress in normal adult mice, and another which elicits progressively growing tumors in the majority of recipients. The tumor incidence, mean tumor size, and tempo of regression were not modified by treatment with anti-IgM. However, whereas tumors induced by the regressor virus were all rejected in 19 NRG-treated and 29 untreated recipients, continued growth was obtained in 2 of 23 B-lymphocyte-deprived mice. Furthermore, in 9 additional mice from this group, apparent rejection was followed by tumor recurrence at the site of the initial tumor. Continued growth was accompanied by widespread metastasis. These tumors were freely transplantable to normal syngeneic recipients. Metastasis and transplantability were also detected in 7 of 24 anti-IgM-treated mice given progressor virus, but were not seen in the control animals. Recurrence and metastasis was obtained despite the presence of high levels of specific cytotoxic T lymphocytes in the spleen. It is concluded that B lymphocytes or their products play an essential role in host protection against MSV-induced tumors.

Animals↗

Determination of [amide-15N]glutamine in plasma with gas chromatography-mass spectrometry.

A gas chromatographic-mass spectrometric method is described for determination of isotopic enrichment in [amide-15N]glutamine. The method involves the isolation of glutamine with an anionic exchange resin and the enzymatic hydrolysis of glutamine with glutaminase. The ammonia derived from the amide group is converted enzymatically to glutamic acid and isotopic enrichment in the latter compound is determined following formation of the N-TFA-glutaminate derivative. The method gave a linear result for [amide-15N]glutamine in the 0.5--21.5 atom % excess range with a mean coefficient of variance of 7.8%. Determination of plasma glutamine by isotope dilution showed a mean recovery of 94%. The appearance of plasma [amide-15N]glutamine was measured in a healthy adult following the intravenous administration of 15NH4Cl (350 mumol). A peak enrichment of 12 atom % excess was observed by 3.5 min.

Ammonia↗