Peptides Abstracts 4

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Carcinogenesis and aging. IV. Effect of low-molecular-weight factors of thymus, pineal gland and anterior hypothalamus on immunity, tumor incidence and life span of C3H/Sn mice.
            (Anisimov et al., 1982) Download
The low-molecular-weight polypeptide factors were obtained from bovine thymus (TF), pineal gland (PF) and anterior hypothalamus (AHF). Both TF and PF administration enhanced the rejection of skin allograft and stimulated the immunological response to sheep erythrocytes in adult CBA mice. Treatment of CBA mice with AHF increased the graft survival and inhibited antibody formation to sheep erythrocytes. Chronic TF or PF administration decreased spontaneous tumor development and prolonged the life span of female C3H/Sn mice. Administration of AHF failed to influence the life span and the tumor incidence of female C3H/Sn mice. The role of immunity and hormonometabolic shifts in mechanisms of both aging and the age-associated increase in cancer incidence are discussed.

Effect of low-molecular-weight factors of thymus and pineal gland on life span and spontaneous tumour development in female mice of different age.
            (Anisimov et al., 1989) Download
Female SHR mice, aged 3.5 or 12 months, were exposed monthly to 5-day long courses of subcutaneous injections of 0.1 mg thymus-derived or pineal gland-derived polypeptide factors (TF and PF, respectively) or 0.9% sodium chloride solution (control). PF treatment increased life span of both young and middle-aged mice by 20% and 17%, respectively, and TF increased the life span only in young mice. Both factors when administered to young mice caused a decrease in both overall tumour incidence and incidence of mammary adenocarcinomas (TF, 1.8-fold decrease; PF, 2.6-fold decrease). TF administration to mature mice did not produce any antitumour effect, whereas PF possessed certain anti-tumour activity, but the response was far less pronounced than in young animals. The results obtained give additional evidence of the geroprotective and anti-tumour effect of thymus and pineal gland-derived peptide factors. The mechanisms of action of TF and PF and perspectives of clinical use of these agents as geroprotectors are discussed.


 

Effect of pineal peptide preparation (epithalamin) on life span and pineal and serum melatonin level in old rats.
            (Anisimov et al., 1992) Download
In the present paper new data on the effect of epithalamin on melatonin synthesis and secretion in young and old rats and on the life span of old female rats are presented. Thus, the data presented allow the suggestions that an ultrashort loop exists between the pineal peptides and indoles, and that the target of pineal peptides is first of all the metabolism of tryptophan into serotonin and its subsequent transformation into melatonin. In old rats the intensity of these reactions is decreased. Administration of epithalamin to old rats was followed by a significant ( p < 0.004) decrease of total tumor incidence (by 1.6times) and a decrease in malignant tumor incidence by 1.8 times ( p < 0.04). Also shown was a decrease in the incidence of pituitary adenomas and mammary tumors . The mechanism of the antitumor and anticarcinogenic effects of epithalamin is thought to involve normalization of a number of hormonal and metabolic disturbances promoting tumor development as well as stimulation of antitumor mediated both by increasing melatonin synthesis and secretion and by direct effects of pineal peptides. The same mechanisms are probably responsible for the aging-preventing effect of the pineal peptide preparation.

Twenty years of study on effects of pineal peptide preparation: epithalamin in experimental gerontology and oncology.
            (Anisimov et al., 1994) Download
In 1973 the first evidence was published that administration of the low-molecular-weight pineal peptide extract (commercial form was named later as Epithalamin) was followed by restoration of the estrus cycle in old female rats with persistent estrus syndrome and by lowering of the threshold of sensitivity of the hypothalamo-pituitary complex to feedback inhibition by estrogens in old animals. Long-term treatment with the preparation prolongs the life span of animals, slows down the aging of the reproductive system, improves the parameters of immune functions and inhibits the development of spontaneous tumors induced by some chemicals or X-irradiation and transplanted tumors.

Pineal peptide preparation epithalamin increases the lifespan of fruit flies, mice and rats.
            (Anisimov et al., 1998) Download
Treatment with pineal peptide preparation epithalamin was followed by the increase of the mean lifespan of female D. melanogaster, SHR mice, C3H/Sn mice and LIO rats by 11-31% (P < 0.05). Ninety percent mortality as well as maximum lifespan were increased in fruit flies, C3H/Sn mice and rats. Mortality rate was decreased by 52% in D. melanogaster, by 52% in rats, by 27% in C3H/Sn mice. It did not change in SHR mice exposed to epithalamin. Treatment with the pineal peptide increased MRDT in flies, C3H/Sn mice and rats. It has been shown that epithalamin increased synthesis and secretion of melatonin in rats and inhibits free radical processes in rats and in D. melanogaster. It is suggested that antioxidative properties of epithalamin lead to increased lifespan of three different animal species.

Effect of Epitalon on biomarkers of aging, life span and spontaneous tumor incidence in female Swiss-derived SHR mice.
            (Anisimov et al., 2003) Download
From the age of 3 months until their natural deaths, female outbred Swiss-derived SHR mice were subcutaneously injected on 5 consecutive days every month with 0.1 ml of normal saline (control) or with 1.0 microg/mouse (approximately 30-40 microg/kg) of tetrapeptide Epitalon (Ala-Glu-Asp-Gly) dissolved in 0.1 ml saline. There were 54 mice in each group. The results of this study show that treatment with Epitalon did not influence food consumption, body weight or mean life span of mice. However, it slowed down the age-related switching-off of estrous function and decreased the frequency of chromosome aberrations in bone marrow cells (by 17.1%, P<0.05). It also increased by 13.3% the life span of the last 10% of the survivors (P<0.01) and by 12.3% the maximum life span in comparison with the control group. We also found that treatment with Epitalon did not influence total spontaneous tumor incidence, but inhibited the development of leukemia (6.0-fold), as compared with the control group. The data obtained suggest a geroprotector activity of Epitalon and the safety of its long-term administration in mice.

[Short peptides stimulate skin cell regeneration during ageing].
            (Chalisova et al., 2014) Download
The actual goal of gerontocosmetology is deal with the research of new effective and harmless low- molecular substances. The influence of LK and AEDG peptides in concentrations 0.05-2.00 ng/ml on organotypic skin cell cultures proliferation in young and old animals were investigated. Peptides stimulated skin fibroblasts proliferation on 29-45% in skin cell cultures of young and old rats. This effect was observed in smaller concentration diapason and level during skin ageing in old cell cultures as compared to young cell cultures. These data open new approach for creation cosmetology substances in the base of LKand AEDG peptides.


 

Increase in lifespan of rats following polypeptide pineal extract treatment.
            (Dilman et al., 1979a) Download
The 20 month-long treatment of female rats with daily doses of 0.1 or 0.5 mg of polypeptide pineal extract (PPE) per animal increased their lifespan by 10 and 25%, respectively, as compared with controls. At the age of 16--18 months, 38% of control rats exhibited persistent disturbances in estral function (constant estrus or repeated pseudogestations), whereas these disorders were observed in 7% of experimental animals only. After administration of PPE to 16--18 month-old female rats checked for sterility by a two-week mating, a second mating period resulted in gestation development in four out of 16 animals and deliveries, accordingly. While chronic treatment with PPE did not affect the rate of neoplasm incidence, the mean age of tumour detection in the control group was 697 days and in experimental groups it was 811 and 868 days, respectively. Certain aspects of the interrelationship of rate of ageing, lifespan and specific age pathology are discussed.

Study of the anti-tumor effect of polypeptide pineal extract.
            (Dilman et al., 1979b) Download
Bovine pineal polypeptide extract (PPE) exerted an anti-tumor effect on mouse-transplantable tumors: mammary cancer (RSM), squamous cell cervical carcinoma (SCC), hepatoma-22a and lympholeukemia LIO-1, and had no effect on Harding-Passey melanoma and leukemia L-1210. It was shown that PPE possessed the ability to decrease the incidence of DMBA-induced mammary adenocarcinomas in rats. The daily administration of 0.5 mg PPE prolonged the life span of rats by 25% and failed to influence spontaneous tumor development. The arguments in favor of a possible mechanism of anti-tumor action of the pineal gland are submitted. It is suggested that the anti-tumor effect of PPE may occur when the syndrome of cancrophilia is induced by tumor transplantation or chemical carcinogens.

Regulatory effect of Epithalon on production of melatonin and cortisol in old monkeys.
            (Goncharova et al., 2001) Download
The effect of Epithalon on melatonin and cortisol secretion in female rhesus monkeys of various ages was evaluated by enzyme immunoassay. Epithalon stimulated evening melatonin production and normalized circadian rhythms of cortisol production in old monkeys.


 

Pineal peptides restore the age-related disturbances in hormonal functions of the pineal gland and the pancreas.
            (Goncharova et al., 2005) Download
The purpose of this research was to study age-related changes in functioning of pineal and pancreatic glands of non-human primates, rhesus monkeys, and to elucidate the possibility of their corrections with the help of epitalon, a synthetic analogue of the pharmacopoeia drug epithalamin. In old (20-27 years) animals, the basal plasma levels of glucose and insulin were found to be higher, while the night melatonin level was lower in comparison with (6-8 years) young animals. After the glucose administration to old monkeys, a larger area under the curve of the plasma glucose response, a reduced glucose 'disappearance' rate, and a reduced insulin peak (5 min after the glucose administration) were observed in comparison with young animals in similar experiments. The epitalon administration to old monkeys caused the decrease in the basal levels of glucose and insulin and the increase in the basal night melatonin level. Additionally, in the case of old monkeys, epitalon decreased the area under the plasma glucose response curve, markedly increased the glucose 'disappearance' rate and normalized the plasma insulin dynamics in response to glucose administration. Yet, it has not affected the hormonal and metabolic changes in young animals. Thus, epitalon is a promising factor for restoring the age-related endocrine dysfunctions of primates.

Effect of peptide preparation epithalamin on circadian rhythm of epiphyseal melatonin-producing function in elderly people.
            (Korkushko et al., 2004) Download
Circadian rhythm of plasma melatonin concentrations in healthy elderly subjects was studied before and after a course treatment with Epithalamin (peptide preparation from the pineal gland). Epithalamin modulated the melatonin-producing function of the pineal gland. During the dark period plasma melatonin concentration increased in subjects with initially lowered activity of the pineal gland, while in subjects with normal epiphyseal function plasma melatonin concentration tended to decrease.

Geroprotective effect of epithalamine (pineal gland peptide preparation) in elderly subjects with accelerated aging.
            (Korkushko et al., 2006) Download
A 12-year randomized clinical study of epithalamine (pineal gland peptide preparation) was carried out in elderly patients with coronary disease and accelerated aging of the cardiovascular system. Long-term treatment with epithalamine decreased the functional age and degree of cardiovascular aging; exercise tolerance increased. After 12 years the number of elderly subjects dead in the group treated by epithalamine was 28% lower than in the control group, despite the same basic therapy. Cardiovascular mortality was 2-fold lower in patients treated by epithalamine; the incidence of cardiovascular failure and respiratory diseases was 2-fold lower in this group. Long-term treatment with epithalamine was associated with a geroprotective effect on the long-term life prognosis in elderly subjects with accelerated aging.

Peptide geroprotector from the pituitary gland inhibits rapid aging of elderly people: results of 15-year follow-up.
            (Korkushko et al., 2011) Download
The paper presents the results of randomized comparative study of the efficiency of peptide geroprotector from the pituitary gland in elderly patients with rapidly aging cardiovascular system. Over three years 39 coronary patients received, in addition to basic therapy, regular courses of epithalamin (peptide drug), while 40 coronary patients (control group) received basic therapy alone. Long-term treatment with epithalamin (6 courses over 3 years) decelerated aging of the cardiovascular system, prevented age-associated impairment of physical endurance, normalized circadian rhythm of melatonin production and carbohydrate and lipid metabolism. A significantly lower mortality in the group of patients treated with epithalamin in parallel with basic therapy also indicated a geroprotective effect of the peptide preparation from the pineal gland.

Effect of epithalamin on circadian relationship between the endocrine function of the thymus and melatonin-producing function of the pineal gland in elderly people.
            (Labunets et al., 2004b) Download
The effect of epithalamin on circadian rhythms of thymic serum factor titers and melatonin concentrations in the blood of elderly people was studied. Course treatment with epithalamin modulated the rhythm of the thymic endocrine function. The increase in the titer of thymic serum factor at night coincided with the increase in blood melatonin concentration and shift of its acrophase to 3.00, which was characteristic of young people. In elderly people with preserved nocturnal peak of the thymic serum factor titer before therapy the nocturnal (3.00) concentration of melatonin was high and did not change after drug therapy. The correlation between melatonin concentration and titer of thymic serum factor increased after epithalamin treatment.


 

Effects of bioactive factors of the pineal gland on thymus function and cell composition of the bone marrow and spleen in mice of different age.
            (Labunets et al., 2004a) Download
The effects of factors from the pineal gland on the titer of thymic serum factor in the supernatant of 3-h thymus stroma cultures, number of stromal precursor fibroblasts and CD4+ cells in the bone marrow, and CD8+ cells in the spleens of adult and old CBA mice were studied in vitro. Epithalamin, Epithalon, and melatonin appreciably increased the titer of thymic serum factor in the supernatant of thymus stroma cultures from mice of different age and increased the percentage of CD4+ cells in the bone marrow suspension from old animals in vitro. The percentage of CD8+ lymphocytes decreased after incubation of splenic cells from old mice with melatonin. The percentage of bone marrow fibroblast precursor cells from adult and old mice did not appreciably change after incubation with the preparations.

Peptidergic regulation of thymocyte differentiation, proliferation, and apoptosis during aging of the thymus.
            (Lin'kova et al., 2011) Download
The effects of T-31, AB-17, and AB-9 peptides on old (passage 8) thymocyte culture were studied. Only AB-9 peptide exhibited a complex geroprotective effect on thymocytes during their aging. Peptide AB-9 stimulated proliferative activity and differentiation of thymocytes and inhibited their apoptosis.

Peptide Regulation of Skin Fibroblast Functions during Their Aging In Vitro.
            (Lin'kova et al., 2016) Download
The effect peptides KE, KED, AED and AEDG on proliferation (Ki-67), regeneration and aging (CD98hc), apoptosis (caspase-3), and extracellular matrix remodeling (MMP-9) in skin fibroblasts during their aging in culture were studied by immunofluorescent confocal microscopy. All studied peptides inhibited MMP-9 synthesis that increases during aging of skin fibroblasts and enhanced the expression of Ki-67 and CD98hc that are less intensively synthesized during cell aging. Peptides AED and AEDG suppressed caspase-dependent apoptosis that increases during aging of cell cultures.


 

[An open clinical trial of cortexin in treatment of brain ischemia].
            (Mashin et al., 2014) Download
OBJECTIVE:  to study cortexin efficacy in treatment of cognitive and affective disorders in brain ischemia (BI) comorbid to arterial hypertension and/or atherosclerosis. MATERIAL AND METHODS:  The results of the all-Russian screening of cortexin efficacy "Cognitive and affective disorders during treatment brain ischemia with cortexin" conducted in 50 000 patients with BI in 70 Russian cities in 2013 were used for an analysis. All patients received cortexin in dose 10 mg/day during 10 days. Patients were examined before treatment and 11 and 30 days after the beginning of treatment. We analyzed treatment results for 500 patients with BI, stage II, mean age 63,7 ± 10,23 years. Treatment efficacy was assessed using 5-score scale of subjective and objective neurological symptoms, the Five words test, the Schulte test, MMSE, Hamilton anxiety scale and a brief version of the Geriatric depression scale. RESULTS AND CONCLUSION:  A decrease or complete regression of focal neurological symptoms, positive changes in indicators of cognitive impairment, normalization of emotional status and decrease in depression level were noted during the treatment.

Natural and synthetic thymic peptides as therapeutics for immune dysfunction.
            (Morozov and Khavinson, 1997) Download
Natural thymic peptides have been isolated from calf thymus by mild acid extraction. Pharmaceutical containing natural peptides (Thymalin) was put into practice as immunocorrector. One of the immunomodulatory molecules (L-Glu-L-Trp) has been isolated from Thymalin by reversed-phase high performance liquid chromatography. Pharmaceutical containing this agent (Thymogen) was designed on the base of synthesized dipeptide. A novel immunomodulatory dipeptide was synthesized and termed Vilon. Both natural and synthetic pharmaceuticals activated T-cell differentiation, T-cell recognition of peptide-MHC complexes, induced the changes in intracellular composition of cyclic nucleotides and cytokine [interleukin (IL-2), interferon (IFN)] excretion of blood lymphocytes. Synthetic dipeptides activated neutrophil chemotaxis and phagocytosis. They had no influence on antioxidant response in thymocytes in comparison with natural peptides. Thymalin and Thymogen were used in persons with chronic pathology and immune dysfunction. The results indicate that thymic peptides participate in the regulating mechanisms of inflammatory processes as cytokine antagonists and show the difference between natural and synthetic products. It is important for the drugs designed to prevent immune dysfunction development.


 

Immunomodulating effects of Vilon and its analogue in the culture of human and animal thymus cells.
            (Sevostianova et al., 2013) Download
We studied molecular mechanisms of immunoprotective effects of two dipeptides, AB-O and R-1, on cultured human and rat thymic cells. Both dipeptides were shown to increase the expression of lymphocyte differentiation marker CD5 in thymic cells. Dipeptide AB-O induced T-cells precursor differentiation towards CD4(+)T-helpers and its effect was weaker than that of dipeptide R-1. Dipeptide R-1 stimulates differentiation of CD5(+) cells to mature T-helpers and cytotoxic CD8(+) T cells and hence can be considered as a bioactive substance possessing immunomodulator and antiallergic activity.

Effect of tetrapeptide cortagen on regeneration of sciatic nerve.
            (Turchaninova et al., 2000) Download
Intramuscular injection of 10 microg/kg cortagen to rats during 10 days after transsection and suturing of the sciatic nerve increased the growth rate and conduction velocity in the regenerating nerve fibers by 27% and 40%, respectively.

Short Biologically Active Peptides as Epigenetic Modulators of Gene Activity
            (Vanyushin and Khavinson, 2016) Download
Short (2–4 aa) peptides investigated increased life span of rats and improved various physiological functions and health status in aged people. Biological activities of peptides are tissue (cell) specific. Peptide bronchogen AlaGluAspLeu (AEDL) regulates synthesis of the Ki67, Mcl-1, p53, CD79, and NOS-3 proteins in cultures of the human bronchoepithelial cells. Bronchogen activates the expression of genes coding for the bronchoepithelial cell differentiation factors Nkx2. 1, SCGB1A1, SCGB3A2, FoxA1, FoxA2

Peptides Restore Functional State of the Kidneys During Cisplatin-Induced Acute Renal Failure.
            (Zamorskii et al., 2015) Download
The effects of polypeptide complex from the kidney and short peptides AED, EDL, and AEDG on renal functions were studied in rats with cisplatin-induced acute renal failure. AED peptide decreased protein excretion and electrolyte concentration in the urine. Polypeptide complex from the kidney and peptides EDL and AEDG normalized diuresis, creatinine concentration in the urine and its excretion, glomerular filtration rate, and absolute resorption of sodium ions and reduced protein concentration in the urine and its excretion, the concentrations of sodium and potassium ions in the urine, and other parameters. EDL peptide produced was potent nephroprotective effect. It is known that the polypeptide complex of the kidney and short peptides restore the expression of signal molecules (marker of functional state of the kidneys), so these peptide substances can have nephroprotective effect during various renal pathologies.

 


References

Anisimov, VN, VK Khavinson, and VG Morozov (1982), ‘Carcinogenesis and aging. IV. Effect of low-molecular-weight factors of thymus, pineal gland and anterior hypothalamus on immunity, tumor incidence and life span of C3H/Sn mice.’, Mech Ageing Dev, 19 (3), 245-58. PubMed: 6752596
Anisimov, VN, et al. (1989), ‘Effect of low-molecular-weight factors of thymus and pineal gland on life span and spontaneous tumour development in female mice of different age.’, Mech Ageing Dev, 49 (3), 245-57. PubMed: 2682058
Anisimov, VN, LA Bondarenko, and VKh Khavinson (1992), ‘Effect of pineal peptide preparation (epithalamin) on life span and pineal and serum melatonin level in old rats.’, Ann N Y Acad Sci, 673 53-57. PubMed: 1485734
Anisimov, VN, VKh Khavinson, and VG Morozov (1994), ‘Twenty years of study on effects of pineal peptide preparation: epithalamin in experimental gerontology and oncology.’, Ann N Y Acad Sci, 719 483-93. PubMed: 8010617
Anisimov, VN, SV Mylnikov, and VK Khavinson (1998), ‘Pineal peptide preparation epithalamin increases the lifespan of fruit flies, mice and rats.’, Mech Ageing Dev, 103 (2), 123-32. PubMed: 9701766
Anisimov, VN, et al. (2003), ‘Effect of Epitalon on biomarkers of aging, life span and spontaneous tumor incidence in female Swiss-derived SHR mice.’, Biogerontology, 4 193-202. PubMed: 14501183
Chalisova, NI, et al. (2014), ‘[Short peptides stimulate skin cell regeneration during ageing].’, Adv Gerontol, 27 (4), 699-703. PubMed: 25946846
Dilman, VM, et al. (1979a), ‘Increase in lifespan of rats following polypeptide pineal extract treatment.’, Exp Pathol (Jena), 17 (9), 539-45. PubMed: 575333
Dilman, VM, et al. (1979b), ‘Study of the anti-tumor effect of polypeptide pineal extract.’, Oncology, 36 (6), 274-80. PubMed: 117414
Goncharova, ND, BK Khavinson, and BA Lapin (2001), ‘Regulatory effect of Epithalon on production of melatonin and cortisol in old monkeys.’, Bull Exp Biol Med, 131 (4), 394-96. PubMed: 11550036
Goncharova, ND, et al. (2005), ‘Pineal peptides restore the age-related disturbances in hormonal functions of the pineal gland and the pancreas.’, Exp Gerontol, 40 (1-2), 51-57. PubMed: 15664732
Korkushko, OV, et al. (2004), ‘Effect of peptide preparation epithalamin on circadian rhythm of epiphyseal melatonin-producing function in elderly people.’, Bull Exp Biol Med, 137 (4), 389-91. PubMed: 15452611
Korkushko, OV, et al. (2006), ‘Geroprotective effect of epithalamine (pineal gland peptide preparation) in elderly subjects with accelerated aging.’, Bull Exp Biol Med, 142 (3), 356-59. PubMed: 17426848
Korkushko, OV, et al. (2011), ‘Peptide geroprotector from the pituitary gland inhibits rapid aging of elderly people: results of 15-year follow-up.’, Bull Exp Biol Med, 151 (3), 366-69. PubMed: 22451889
Labunets, IF, GM Butenko, and VKh Khavinson (2004a), ‘Effects of bioactive factors of the pineal gland on thymus function and cell composition of the bone marrow and spleen in mice of different age.’, Bull Exp Biol Med, 137 (5), 510-12. PubMed: 15455131
Labunets, IF, et al. (2004b), ‘Effect of epithalamin on circadian relationship between the endocrine function of the thymus and melatonin-producing function of the pineal gland in elderly people.’, Bull Exp Biol Med, 137 (5), 507-9. PubMed: 15455130
Lin’kova, NS, et al. (2011), ‘Peptidergic regulation of thymocyte differentiation, proliferation, and apoptosis during aging of the thymus.’, Bull Exp Biol Med, 151 (2), 239-42. PubMed: 22238759
Lin’kova, NS, et al. (2016), ‘Peptide Regulation of Skin Fibroblast Functions during Their Aging In Vitro.’, Bull Exp Biol Med, 161 (1), 175-78. PubMed: 27259496
Mashin, VV, et al. (2014), ‘[An open clinical trial of cortexin in treatment of brain ischemia].’, Zh Nevrol Psikhiatr Im S S Korsakova, 114 (9), 49-52. PubMed: 25403301
Morozov, VG and VK Khavinson (1997), ‘Natural and synthetic thymic peptides as therapeutics for immune dysfunction.’, Int J Immunopharmacol, 19 (9-10), 501-5. PubMed: 9637345
Sevostianova, NN, et al. (2013), ‘Immunomodulating effects of Vilon and its analogue in the culture of human and animal thymus cells.’, Bull Exp Biol Med, 154 (4), 562-65. PubMed: 23486604
Turchaninova, LN, et al. (2000), ‘Effect of tetrapeptide cortagen on regeneration of sciatic nerve.’, Bull Exp Biol Med, 130 (12), 1172-74. PubMed: 11276314
Vanyushin, BF and V Kh Khavinson (2016), ‘Short Biologically Active Peptides as Epigenetic Modulators of Gene Activity’, Epigenetics-A Different Way of Looking at Genetics (Springer), 69-90.
Zamorskii, II, et al. (2015), ‘Peptides Restore Functional State of the Kidneys During Cisplatin-Induced Acute Renal Failure.’, Bull Exp Biol Med, 159 (6), 736-39. PubMed: 26515176