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Aqueous garlic extract inhibits protamine sulfate-induced bladder damage.

This morphological and biochemical study aims to investigate the antioxidant effects of chronic administration of aqueous garlic extract (AGE) on protamine sulfate (PS)-induced bladder injury. Wistar albino female rats were catheterized and intravesically infused with phosphate-buffered solution (control group) or PS (PS group) dissolved in phosphate-buffered solution. In the PS + AGE group after the PS instillation, AGE (1 ml/kg, i.p., corresponding to 250 mg/kg) was injected intraperitoneally for 3 days. Bladder morphology was investigated by light and scanning electron microscopy. Tissue samples were also obtained to determine bladder malondialdehyde (MDA) and glutathione levels. In the PS group, ulcerated areas, an irregular mucus layer, inflammatory cell infiltration and an increased number of mast cells were observed. In the PS + AGE group a relatively normal urothelial topography, glycosaminoglycan layer and a decreased number of mucosal mast cells and inflammatory cells were observed. Increased MDA levels as a result of PS induction led us to propose that free radicals may have a critical role in this injury. The significant decrease in MDA and increase in glutathione levels in the PS + AGE group was in accordance with morphological findings. Based on the results, AGE treatment significantly prevented PS-induced degenerative morphological and biochemical changes of urinary bladder mucosa.

Animals↗

The genes for protamine 1 and 2 (PRM1 and PRM2) and transition protein 2 (TNP2) are closely linked in the mammalian genome.

The genes for two protamines (PRM1 and PRM2) and for two transition proteins (TNP1 and TNP2) have been characterized in several mammalian species. In the human, boar, and bull, the genes for PRM1, PRM2, and TNP2 are closely linked over a stretch of DNA 13-15 kb long. Although similar data are not yet available for the mouse and rat, our results suggest that the three genes are similarly linked in these species. The gene for TNP1 in all species studied is located on another chromosome.

Animals↗

Neutralization of DHG, a new depolymerized holothurian glycosaminoglycan, by protamine sulfate and platelet factor 4.

The neutralization of depolymerized holothurian glycosaminoglycan (DHG), unfractionated heparin (UFH), and low-molecular-weight heparin (LMWH) by protamine sulfate (PS) or platelet factor 4 (PF4) was studied. In in vitro studies, the prolongation of thrombin clotting time (TCT) by these glycosaminoglycans was completely neutralized by PS, whereas activated partial thromboplastin time (APTT) was relatively resistant to neutralization. In rats, prolongation of bleeding time by DHG was neutralized by PS with concomitant normalization of TCT ex vivo. Heparin-cofactor-II-dependent antithrombin activity of DHG or UFH was neutralized by PF4 at a high concentration to the same extent; however, prolongation of the APTT by DHG was more resistant to neutralization by PF4 at a physiological plasma level than that by UFH. In conclusion, since the prolongation of bleeding time by DHG was neutralized by PS with concomitant normalization of TCT, and since PF4 neutralized the antithrombin activity of DHG, these proteins may be useful as antidotes for DHG to prevent bleeding in case of an overdose.

Animals↗

Effects of glycosaminoglycans and protamine chloridrate on platelet aggregation induced by collagen and thrombin.

The effects of heparin (HE), dermatan sulfate (DS), heparan sulfate (HS) and protamine chloridrate (PC) on platelet aggregation were studied. Both PC and the three glycosaminoglycans (GAGs) did not influence collagen-induced platelet aggregation. In contrast, all the tested GAGs blocked thrombin-induced platelet aggregation. HE and HS were equivalent and very effective, while DS was also but to a lesser extent. This could be because HE and HS act via both antithrombin III and heparin-cofactor II, whereas DS exerts its action on the latter only. PC, too, inhibited, in a dose-dependent fashion, thrombin-induced platelet aggregation, probably by competing with the thrombin affinity binding sites on the platelet surface. When the GAGs were tested together with PC, HE was shown to be the most effective: on a weight-for-weight basis, an identical amount of PC was unable to counteract the inhibitory effect of HE, while it partially reversed those of DS and HS. A full reversal of the inhibitory effect of DS and HS was never observed, in spite of adding increasing amounts of PC. It seems likely that plasma components may preserve the bindings of such GAGs with their cofactors.

Collagen↗

Experimental colon neoplasia enhanced by extract of sperm (protamine).

An experimental study of colorectal tumorigenesis in inbred rats given the potent carcinogen dimethylhydrazine has shown that protamine, given before any tumours had developed, significantly enhanced the development of large bowel tumours (100% incidence). Those animals having no adjuvant therapy or who received either heparin or 5-fluorouracil (alone or in combination) had an approximate 50% incidence of such tumours.

Adenocarcinoma↗

Cytoplasmic localization during storage and translation of the mRNAs of transition protein 1 and protamine 1, two translationally regulated transcripts of the mammalian testis.

During spermatogenesis in mammals, the transcripts of transition protein 1 (TP 1) and protamine 1 (Prm 1) are under translational regulation. Following their transcription in round spermatids, the mRNAs for TP 1 and Prm 1 are stored in the cytoplasm from 3-7 days before being translated towards the end of spermatogenesis. To test the hypothesis that the inactivation or activation of transcripts during spermiogenesis could be mediated by mRNA compartmentalization in the cytoplasm of spermatids, light and electron microscopy were used to localize, by in situ hybridization, the cellular and subcellular sites of stored and translated mRNAs for these two testis-specific transcripts. During early spermiogenesis (before step 7) nuclear transcripts of both TP 1 and Prm 1 were seen. After step 7 the TP 1 and Prm 1 mRNAs were only detected in the cytoplasm. Throughout spermiogenesis the cytoplasmic mRNAs were not localized to any membrane-bound organelles such as the endoplasmic reticulum or mitochondria or to non-membrane-bound structures such as the chromatoid body. These studies demonstrate that the translational arrest of the TP 1 and Prm 1 mRNAs is not primarily controlled by compartmentalized storage in the cytoplasm of spermatids. Moreover, when translation of these mRNAs occurs in elongated spermatids, the mRNAs are present throughout the cytoplasm.

Animals↗

Purification and properties of an aminopeptidase from a protamine-degrading marine bacterium.

A protamine-degrading marine bacterium was isolated from marine soil and identified as Aeromonas salmonicida subsp. based on its taxonomical characteristics. An alanine-specific aminopeptidase, called aminopeptidase K, from an extract of the strain was purified and characterized. The aminopeptidase K was purified about 80-fold by fractionation with ammonium sulfate and column chromatography on QA-52 cellulose, Phenyl Superose and Superose 12. The purified enzyme is composed of 6 subunits of 86 kDa with a molecular mass of 520 kDa according to gel filtration and SDS-PAGE. The N-terminal sequence of the enzyme was H.Gly-Gln-Gln-Pro-Gln-Ile-Lys-Try-Tyr-His-Asp-Tyr-Asp-Ala-Pro-Asp-Tyr-Ty r- Ile-Thr-. It is inhibited by monoiodoacetate, N-ethylmaleimide, and puromycin. The Michaelis constant (K(m)) and the maximal rate of hydrolysis (Vmax) were, respectively, 0.28 mM and 49.4 mumol/min/mg for the L-Ala-beta-naphthylamide substrate. The optimum pH and optimum temperature were 6.5 and 45 degrees C, respectively. The purified enzyme was highly specific to L-Ala-beta-naphthylamide.

Aeromonas↗

Characterization of a gene encoding a basic protein of the spermatid nucleus, TNP2, and its close linkage to the protamine genes in the bull.

During elongation and condensation of the spermatid nucleus, histones are replaced by spermatid-specific transition proteins (TNP). TNP1 is well characterized at the cDNA and at the genomic level and was found to be highly conserved during mammalian evolution (similarity between 83 to 98%). We here describe for the first time the nucleotide sequence and organization of the gene for TNP2. The gene was isolated from a bull cosmid library and was found to contain a single intron of 910 bp. The coding sequence consists of 390 bp and has a similarity of about 70% to that of the TNP2 cDNAs of mouse and rat. At the basis of amino-acid sequences, the bull TNP2 is 14 and 15 amino acids longer than that of mouse and rat, respectively, and the similarity is only 45% between bull and mouse and 42% between bull and rat. However, the evolutionary divergence has not occurred at the cost of basic amino acids which are of functional importance in DNA-protein interaction in the condensing spermatid nucleus. The TNP2 gene is closely linked to the protamine genes in the bull genome.

Aging↗

The effect of protamine on serum complement activity in aminonucleoside nephropathy.

Combined administration of protamine sulfate (Ps) and aminonucleoside (AN) to rats causes severe nephrotic syndrome and, histologically, focal glomerular sclerosis. These changes are more distinct than those produced by AN alone. While the exact mechanism of Ps acting on AN nephropathy is not known, marked hypocomplementemia was observed regularly after the Ps + AN injection, suggesting that complement is activated and consumed in the kidney. It is suggested that complement may play an important role in AN + Ps nephropathy.

Animals↗

Lipid-protamine-DNA-mediated antigen delivery.

The development of novel 'new generation' vaccine systems that is based on proteins, peptides or DNA is of great current interest. However, due to the lower efficiencies of these new generation vaccines, they are seldomly used alone. Rather, their formulations often contain adjuvants, either to enhance the immune responses or to reduce dosing. The present chapter will provide a brief overview of the recent advances in peptide-based cancer vaccine adjuvants, focusing mainly on Liposome-Protamine-DNA (LPD) nanoparticle-mediated antigen delivery.

Adjuvants, Immunologic↗

Improved postprandial glycemic control during treatment with Humalog Mix25, a novel protamine-based insulin lispro formulation. Humalog Mix25 Study Group.

OBJECTIVE: Humalog Mix25 is a manufactured premixed insulin formulation containing insulin lispro and a novel insulin lispro-protamine formulation (NPL) in a ratio of 25:75%. The objective of this study was to compare Humalog Mix25 to human insulin 30/70 (30% regular insulin/70% NPH) with respect to glycemic control. RESEARCH DESIGN AND METHODS: Humalog Mix25 was compared with human insulin 30/70 in 89 individuals with type 2 diabetes during a 6-month randomized open-label two-period crossover study. Each insulin was administered twice daily, before the morning and evening meals. Information regarding self-monitored blood glucose (BG), hypoglycemic episodes (hypoglycemic signs or symptoms or BG < or = 3.0 mmol/l), insulin dose, and HbA1c was collected. RESULTS: Treatment with Humalog Mix25 resulted in better postprandial glycemic control after the morning and evening meals compared with treatment with human insulin 30/70. Overall glycemic control and the incidence of hypoglycemia were comparable between the treatments. CONCLUSIONS: In comparison to treatment with human insulin 30/70, twice daily administration of Humalog Mix25 resulted in improved postprandial glycemic control, similar overall glycemic control, and the convenience of dosing immediately before meals.

Blood Glucose↗

Decondensation of human sperm nuclei and HP1 protamine degradation from normospermia and asthenospermia in Xenopus egg extracts.

The process of human sperm decondensation has been studied in vitro in cytoplasmic extracts prepared from unfertilized Xenopus laevis eggs. The chromatin decondensation-recondensation cycle was divided into four stages according to chromatin appearance. Spermatozoa from normospermia and asthenospermia were evaluated according to their capacity to reach these stages, and their DNA integrity was assessed by acridine orange (AO) staining. We observed a significant difference between normospermia and asthenozoospermia in the ability to achieve the cycle of chromatin decondensation-recondensation. These results correlated with AO staining. The role of human protamine 1 degradation in the decondensation process was evaluated by immunostaining. It was found not to be a prerequisite for the earlier stage of chromatin decondensation and it was not implied in the latest stages of pronuclear development.

Acridine Orange↗

Adjuvant and suppressor activity of the polycation protamine hydrochloride in the primary immune response of mice.

The effect of protamine hydrochloride (PH), a polycation, on the primary immune response of mice to sheep red blood cells (SRBC) was studied. Like other adjuvants, PH enhances or depresses the immune response depending on the time of injection of PH in relation to the antigenic challenge. The suppressive effect of PH correlated with the appearance of "activated" macrophages in the peritoneal cavity. A second injection of PH together with antigen abolished this suppressive effect. Transfer of "activated" macrophages resulted in an enhanced immune response when antigen was administered at the time of transfer, and in a depressed response when the antigen was given 1 or 2 days after transfer. It is concluded that the duration of the influence of "activated" macrophages on T or B cells determines the subsequent immune reactivity. It is discussed that the basic mechanism of action of PH is its ability to release lysosomal enzymes from macrophages. The mechanism by which these enzymes amplify the immune response remains to be clarified.

Adjuvants, Immunologic↗

[Prevention of negative hemodynamic effects of protamine sulfate by simultaneous infusion of microdoses of adrenaline].

Two groups of coronary patients subjected to revascularization of the myocardium were examined in order to detect the negative hemodynamic effects of protamine sulfate (PS) and the possibility of their correction by simultaneous infusion of adrenaline microdoses. In group 1 (27 pts.) heparin was neutralized by infusion of PS alone (6 mg/kg) and in group 2 (27 pts.) by simultaneous infusion of PS and adrenaline (15 ng/kg/min). The functions of the right and left ventricles were evaluated by catheterization of the pulmonary artery by a Swan-Ganz catheter and transesophageal Doppler echocardiography. These methods provided volume and velocity characteristics of the right and left compartments of the heart. PS deteriorated the systolic function of the right ventricle, particularly in patients with initial dysfunction of the right heart. Infusion of adrenaline simultaneously with PS leveled its negative effects, thus preventing myocardial dysfunction.

Adrenergic Agonists↗

Enzyme potentiated hyposensitization: IV. effect of protamine on the immunological behavior of beta glucuronidase in mice and patients with hay fever.

The ability of beta glucuronidase and a small dose of antigen to modify the anaphylactic reaction of previously sensitized mice has been further investigated. Protamine has an important effect on the immunological behavior of the enzyme. A trial on hay fever patients shows that the results in mice are relevant and that the method can produce significant clinical hyposensitization.

Animals↗

Improved glycaemic control with the use of self-prepared mixtures of insulin lispro and insulin lispro protamine suspension in patients with types 1 and 2 diabetes.

A total of 166 patients (102 type 1, 64 type 2) were randomised to insulin lispro (LP) combined with insulin lispro protamine suspension (NPL), an intermediate-acting formulation of LP, or to regular human insulin (HR) combined with human NPH insulin (NPH) in this open-label, parallel study. Insulin doses were similar at endpoint. Blood glucose (BG) measurements (before and two hours after meals, bedtime, 3 a.m.)(mmol/l) were lower with LP/NPL two hours after breakfast (8.84 +/- 0.32 vs 10.29 +/- 0.41, p < 0.001), before lunch (6.21 +/- 0.20 vs 7.10 +/- 0.31, p = 0.016), two hours after the evening meal (10.18 +/- 0.36 vs 7.86 +/- 0.28, p < 0.0.001), and at bedtime (7.85 +/- 0.28 vs 9.43 +/- 0.40, p = 001). HbA1c was lower for LP/NPL at endpoint (7.54 +/- 0.11% vs 7.92 +/- 0.10%, p = 0.019). There was no difference in hypoglycaemia or insulin antibody levels. LP/NPL resulted in better glycaemic control than HR/NPH without increasing the risk of hypoglycaemia.

Adolescent↗

Studies on a protamine (galline) from fowl sperm. 2. The amino acid sequences of two components of galline.

Galline, a protamine from rooster sperm nuclei, has already been separated into its components, G-I-G-VIII. The amino acid composition of the homogeneously purified fraction G-I is determined decisively to be Arg11, Ser2, Gly3, Val1 and Tyr2, and the molecular weight is 2,908 as hydrochloride. The complete amino acid sequence was deduced as follows from the results of analyses of the tryptic and chymotryptic peptides: H-Ser-Gly-Gly-Val-Arg-Arg-Arg-Arg-Tyr-Gly-Ser-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Tye-OH. Another purified fraction, G-V, has the amino acid composition, Arg24, Thr1, Ser8, Gly3, Ala2, Pro2 and Tyr2 as described previously, and the molecular weight 6,274 as hydrochloride. The complete amino acid sequence was established by combining analytical results of tryptic, chymotryptic and thermolysin peptides as well as those of carboxypeptidase and leucine aminopeptidase digestion as follows: H-Ala-Arg-Tyr-Arg-Ser-Gly-Arg-Ser-Arg-Ser-Arg-Arg-Thr-Arg-Arg-Arg-Arg-Ser-Pro-Arg-Ser-Arg-Gly-Arg-Ser-Pro-Arg-Arg-Arg-Arg-Ser-Arg-Arg-Arg-Arg-Arg-Tyr-Gly-Ser-Ala-Arg-Arg-OH.

Amino Acid Sequence↗