Overexpression of the lat gene in Nocardia lactamdurans from strong heterologous promoters results in very high levels of lysine-6-aminotransferase and up to two-fold increase in cephamycin C production

2000 ◽  
Vol 53 (3) ◽  
pp. 282-288 ◽  
Author(s):  
V. K. Chary ◽  
J. L. de la Fuente ◽  
A. L. Leitão ◽  
P. Liras ◽  
J. F. Martín
Keyword(s):  
Fold Increase ◽  
Cephamycin C ◽  
Nocardia Lactamdurans ◽  
Very High

Cloning and functional expression of an MIP (AQP0) homolog from killifish (Fundulus heteroclitus) lens

2001 ◽  
Vol 281 (6) ◽  
pp. R1994-R2003 ◽  
Author(s):  
Leila V. Virkki ◽  
Gordon J. Cooper ◽  
Walter F. Boron
Keyword(s):  
Water Permeability ◽  
Fundulus Heteroclitus ◽  
Water Channel ◽  
Fold Increase ◽  
Functional Expression ◽  
Xenopus Laevis Oocytes ◽  
Lens Fiber ◽  
Fiber Cells ◽  
Aqp1 Expression ◽  
Very High

The major intrinsic protein (MIP) of lens fiber cells is a member of the aquaporin (AQP) water channel family. The protein is expressed at very high levels in lens fiber cells, but its physiological function is unclear. By homology to known AQPs, we have cloned a full-length cDNA encoding an MIP from the lens of killifish ( Fundulus heteroclitus). The predicted protein (263 amino acids; GenBank accession no. AF191906 ) shows 77% identity to amphibian MIPs, 70% identity to mammalian MIPs, and 46% identity to mammalian AQP1. Expression of MIPfun in Xenopus laevis oocytes causes an ∼40-fold increase in oocyte water permeability. This stimulation is comparable to that seen with AQP1 and substantially larger than that seen with other MIPs. The mercurials HgCl2 and p-chloromercuribenzenesulfonate inhibit the water permeability of MIPfun by ∼25%. MIPfun is not permeable to glycerol, urea, or formic acid but is weakly permeable to CO2.

High levels of androgens cause chondrocyte accumulation and loss of smooth muscle in the mouse penile body†

2020 ◽  
Vol 102 (6) ◽  
pp. 1225-1233
Author(s):  
Deepak S Hiremath ◽  
Elizabeth C Geerling ◽  
Lan Hai ◽  
Prema Narayan
Keyword(s):  
Smooth Muscle ◽  
Structural Changes ◽  
Morphological Changes ◽  
Serum Testosterone ◽  
Fold Increase ◽  
Luteinizing Hormone Receptor ◽  
Corpora Cavernosa ◽  
Structural Abnormalities ◽  
Very High ◽  
Androgen Levels

Abstract Androgens are essential for penile development and for maintaining penile structural and functional integrity. Loss of androgen levels or function results in a decrease in smooth muscle content, accumulation of adipocytes in the corpora cavernosa, and inhibition of erectile function. Our previous studies with a mouse model (KiLHRD582G) of constitutive luteinizing hormone receptor activity also showed structural abnormalities in the penis caused by a decrease in smooth muscle content, accumulation of chondrocytes, and sexual dysfunction. As KiLHRD582G mice exhibit very high levels of testosterone at all postnatal ages, the goal of this study was to determine if the elevated androgen levels were responsible for the morphological changes in the penis. Implantation of testosterone capsules in wild-type mice at neonatal (2 weeks) and postpubertal (5 weeks) ages resulted in the accumulation of chondrocytes in the corpora cavernosa of the adult animals. Mice implanted with testosterone capsules at 2 weeks of age exhibited a 4-fold increase in serum testosterone with a 1.5-fold loss of smooth muscle at 24 weeks of age. Collagen content was unchanged. Only 57% of testosterone implanted mice were fertile at 24 weeks of age. Mice implanted with testosterone capsules at 5 weeks of age showed no decrease in smooth muscle content at 24 weeks, although serum testosterone levels were elevated 5-fold. Implantation with dihydrotestosterone also resulted in chondrocyte accumulation and a 2-fold loss in smooth muscle content. Together, these studies demonstrate that supraphysiological levels of androgens cause structural changes in the penile corpora cavernosa and impair fertility.

scholarly journals Transgenic mice expressing the human ornithine decarboxylase gene under the control of mouse metallothionein I promoter

1996 ◽  
Vol 314 (2) ◽  
pp. 405-408 ◽  
Author(s):  
Leena ALHONEN ◽  
Sami HEIKKINEN ◽  
Riitta SINERVIRTA ◽  
Maria HALMEKYTÖ ◽  
Pekka ALAKUIJALA ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Ornithine Decarboxylase ◽  
Mammalian Cells ◽  
Fold Increase ◽  
Transgenic Animals ◽  
Decarboxylase Activity ◽  
Transgenic Mouse Line ◽  
Regulatory Machinery ◽  
Dividing Cells ◽  
Very High

We have generated a transgenic mouse line harbouring the human ornithine decarboxylase gene under the control of mouse metallothionein I promoter. Even in the absence of an exposure to heavy metals, ornithine decarboxylase was over-expressed in heart, testis, brain, and especially in liver, of the transgenic animals. An exposure of the transgenic mice to zinc further enhanced the enzyme activity to a level which in liver represented up to 8000-fold increase in comparison with non-transgenic animals. The striking stimulation of liver ornithine decarboxylase activity upon treatment of the transgenic mice with zinc was accompanied by a nearly 150-fold increase in the hepatic putrescine content as compared with similarly treated non-transgenic animals. Even though the liver putrescine concentration reached that of spermidine and spermine in the transgenic animals, the contents of the higher polyamines only transiently increased upon zinc administration and then returned to the basal level. These findings once again indicate that mammalian cells possess extremely powerful regulatory machinery to prevent an over-accumulation of spermidine and spermine in non-dividing cells, and that very high tissue putrescine concentrations can be tolerated, at least for periods of a few days, with seemingly no phenotypic changes.

TYPE 1 DIABETES IN PREGNANCY; INFLUENCES ON MOTHER AND FETUS

2009 ◽  
Vol 20 (1) ◽  
pp. 17-47 ◽  
Author(s):  
SCOTT M NELSON ◽  
ROBERT S LINDSAY
Keyword(s):  
Type 1 Diabetes ◽  
Perinatal Mortality ◽  
Glucose Monitoring ◽  
Perinatal Outcomes ◽  
Fold Increase ◽  
Gradual Improvement ◽  
Management Of Complications ◽  
The Uk ◽  
Very High

Type 1 diabetes complicates around 1 in 200 to 300 pregnancies in the United Kingdom. Historically maternal type 1 diabetes carried very high risks for mother and child. Introduction of insulin led to an immediate, marked decline in the previously very high rates of maternal mortality; in contrast an improvement in perinatal outcomes occurred more slowly but was nevertheless dramatic. This is strikingly demonstrated by the temporal decline in perinatal mortality in offspring of mothers with type 1 diabetes which was virtually universal before use of insulin in the 1920's, likely remained in excess of 20% even in the 1960's and fell to under 4% by the 1990's. The reasons for this more gradual improvement in perinatal outcomes cannot be defined with precision but will have been influenced by improved glycaemic management with use of intensive, multiple dose insulin treatment and home glucose monitoring; improvements in obstetric and neonatal management, and better management of complications of diabetes before and during pregnancy. In 1989 the St Vincent declaration proposed that pregnancy outcomes in women with type 1 diabetes should approximate those of the non-diabetic population. While the long term improvements in fetal outcomes have been dramatic, contemporary surveys confirm a persistent doubling or more of rates of congenital anomaly and a three to four fold increase in perinatal mortality in the UK and other European countries which will require further clinical innovation to overcome.

Δ-1-Piperideine-6-carboxylate dehydrogenase, a new enzyme that forms α-aminoadipate in Streptomyces clavuligerus and other cephamycin C-producing actinomycetes

1997 ◽  
Vol 327 (1) ◽  
pp. 59-64 ◽  
Author(s):  
Juan L. de la FUENTE ◽  
Angel RUMBERO ◽  
Juan F. MARTÍN ◽  
Paloma LIRAS
Keyword(s):  
Dehydrogenase Activity ◽  
Maximal Activity ◽  
Streptomyces Clavuligerus ◽  
Electron Acceptors ◽  
Native Protein ◽  
Cephamycin C ◽  
Second Stage ◽  
Streptomyces Cattleya ◽  
Nocardia Lactamdurans ◽  
Blue Sepharose

Δ-1-Piperideine-6-carboxylate (P6C) dehydrogenase activity, which catalyses the conversion of P6C into α-aminoadipic acid, has been studied in the cephamycin C producer Streptomyces clavuligerus by both spectrophotometric and radiometric assays. The enzyme has been purified 124-fold to electrophoretic homogeneity with a 26% yield. The native protein is a monomer of 56.2 kDa that efficiently uses P6C (apparent Km 14 μM) and NAD+ (apparent Km 115 μM), but not NADP+ or other electron acceptors, as substrates. The enzyme activity was inhibited (by 66%) by its end product NADH at 0.1 mM concentration. It did not show activity towards pyrroline-5-carboxylate and was separated by Blue-Sepharose chromatography from pyrroline-5-carboxylate dehydrogenase, an enzyme involved in the catabolism of proline. P6C dehydrogenase reached maximal activity later than other early enzymes of the cephamycin pathway. The P6C dehydrogenase activity was decreased in ammonium (40 mM)-supplemented cultures, as was that of lysine 6-aminotransferase. P6C dehydrogenase activity was also found in other cephamycin C producers (Streptomyces cattleya and Nocardia lactamdurans) but not in actinomycetes that do not produce β-lactams, suggesting that it is an enzyme specific for cephamycin biosynthesis, involved in the second stage of the two-step conversion of lysine to α-aminoadipic acid.

scholarly journals Inducing Effect of Diamines on Transcription of the Cephamycin C Genes from the lat and pcbAB Promoters inNocardia lactamdurans

1999 ◽  
Vol 181 (8) ◽  
pp. 2379-2384 ◽  
Author(s):  
Ana Lucia Leitão ◽  
Francisco J. Enguita ◽  
Juan Luis De La Fuente ◽  
Paloma Liras ◽  
Juan F. Martin
Keyword(s):  
Cell Growth ◽  
Antibiotic Production ◽  
Protein A ◽  
Transcriptional Level ◽  
Intracellular Accumulation ◽  
Low Resolution ◽  
Cephamycin C ◽  
Nocardia Lactamdurans ◽  
Shake Flasks ◽  
Acid Precursor

ABSTRACT The diamines putrescine, cadaverine, and diaminopropane stimulate cephamycin biosynthesis in Nocardia lactamdurans, in shake flasks and fermentors, without altering cell growth. Intracellular levels of the P7 protein (a component of the methoxylation system involved in cephamycin biosynthesis) were increased by diaminopropane, as shown by immunoblotting studies. Lysine-6-aminotransferase and piperideine-6-carboxylate dehydrogenase activities involved in biosynthesis of the α-aminoadipic acid precursor were also greatly stimulated. The diamine stimulatory effect is exerted at the transcriptional level, as shown by low-resolution S1 protection studies. The transcript corresponding to the pcbAB gene and to a lesser extent also the lat transcript were significantly increased in diaminopropane-supplemented cultures, whereas transcription from the cefD promoter was not affected. Coupling of the lat and pcbABpromoters to the reporter xylE gene showed that expression from the lat and pcbAB promoters was increased by addition of diaminopropane in Streptomyces lividans. Intracellular accumulation of diamines in Nocardia may be a signal to trigger antibiotic production.

Parathyroid, renal, and skeletal responses to induced hypocalcemia in the dog

1982 ◽  
Vol 242 (5) ◽  
pp. E287-E291
Author(s):  
J. Fox ◽  
H. Heath
Keyword(s):  
Parathyroid Hormone ◽  
Urinary Excretion ◽  
Infusion Rate ◽  
Elevated Level ◽  
Fold Increase ◽  
Conscious Dogs ◽  
Clamp Technique ◽  
Very High ◽  
Immunoreactive Parathyroid Hormone

This study was designed to determine 1) whether the plasma immunoreactive parathyroid hormone (IPTH) response to acutely attained, constant (8 h) hypocalcemia is biphasic, and 2) if so, how kidney and bone respond to these changing plasma IPTH levels. We initiated constant hypocalcemia (decrement of Ca, 1.7 mg/dl) in six conscious dogs using the "calcium clamp" technique. Plasma IPTH concentrations increased maximally (fivefold) within 15 min and then decreased gradually over 1 h to a constant, but still elevated level (3.2-fold increase). Urinary excretion of phosphate and hydroxyproline increased more slowly, reaching plateaus at 1.75 h (76% increase) and 5.5 h (70% increase), respectively. The EGTA infusion rate required to maintain constant hypocalcemia was virtually constant (85 +/- 9 mumol.kg-1.h-1) after 20 min and corresponded to skeletal release of about 80 mg Ca.kg-1.day-1. The contribution of the kidney in conserving filtered calcium was relatively minor (2.0 +/- 0.5 mumol.kg-1.h-1). These data demonstrate that the parathyroid response to acute, constant hypocalcemia is biphasic and is temporally divergent from the uniphasic phosphaturic and hydroxyprolinuric responses. The ensuing increased skeletal release of calcium is very high and is maximal within minutes.

Translocation t(11;14)(q13;q32) is the hallmark of IgM, IgE, and nonsecretory multiple myeloma variants

Blood ◽  
2003 ◽  
Vol 101 (4) ◽  
pp. 1570-1571 ◽  
Author(s):  
Hervé Avet-Loiseau ◽  
Richard Garand ◽  
Laurence Lodé ◽  
Jean-Luc Harousseau ◽  
Régis Bataille
Keyword(s):  
Multiple Myeloma ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Fold Increase ◽  
Major Interest ◽  
High Incidence ◽  
Very High

In an attempt to address the issue of cytogenetic features of multiple myeloma (MM) variants, we have analyzed a series of 8 IgM, 9 IgD, 2 IgE, and 14 nonsecretory (NS) MM cases using fluorescence in situ hybridization. A very high incidence (83%) of t(11;14)(q13;q32) was detected in the IgM (7 of 8), IgE (2 of 2), and NS (11 of 14) MM cases, but not in the IgD cases (2 of 9). Of note, no t(4;14) was observed in this cohort of patients. This increased incidence of t(11;14) was associated with 2 dominant features in these variants, namely, a “lymphoplasmacytic” presentation mainly in IgM MM and a lower secreting capacity in the others, 2 features previously associated with t(11;14). Of major interest, t(11;14) was never observed in Waldenström macroglobulinemia or in IgG/IgA “lymphoplasmacytic” lymphomas. Thus, for unknown reasons, t(11;14) is the hallmark of IgM, IgE, and NS MM, (but not IgD MM), with a 5-fold increase of its incidence compared to that of IgG and IgA MM.

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