Ursonic acid attenuates spermatogenesis in oligozoospermia mice through inhibiting ferroptosis.

Ursonic acid (UNA) is a natural pentacyclic triterpene found in some medicinal plants and foods. The reproductive protective effect of UNA was evaluated in a mouse model of oligozoospermia induced by busulfan (BUS) at 30 mg/kg b.w.. The mice were initially divided into groups with UNA concentrations of 10, 30, 50, 100 mg/kg. Subsequently, based on sperm parameters, the optimal concentration of 50 mg/kg was identified. As a control, an additional group was supplemented with ursolic acid at a concentration of 50 mg/kg. The results indicated that BUS caused the loss of spermatogenic cells in testis, the decrease of sperm in epididymis, the disorder of testicular cytoskeleton, the decrease of serum sex hormones such as testosterone which induced an increase in feedback of androgen receptor and other testosterone-related proteins, the increase of malondialdehyde and reactive oxygen species levels and the increase of ferroptosis in testis while UNA successfully reversed these injuries. High-throughput sequencing revealed that UNA administration significantly upregulated the expression of genes associated with spermatogenesis, such as Tnp1, Tnp2, Prm1, among others. These proteins are crucial in the histone to protamine transition during sperm chromatin remodeling. Network pharmacology analysis reveals a close association between UNA and proteins related to the transformation of histones to protamine. Molecular docking studies reveal that UNA can interact with the ferroptosis-inhibiting gene SLC7A11, thereby modulating ferroptosis. Taken together, UNA alleviated BUS-induced oligozoospermia by regulating hormone secretion, mitigating oxidative stress and promoting recovery of spermatogenesis by inhibiting the ferroptosis.

Novel peptides as potential treatment of systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by a loss of immunologic tolerance, production of auto-antibodies, and inflammatory damage in multiple organs. We have tested the effect of anti-inflammatory peptide, a H2A histone fragment, termed IIIM1, on MRL/lpr mice, animal model of SLE. Oral administration of IIIM1 at early stage of disease caused reduction in proteinuria and serum anti-dsDNA antibodies. Starting the treatment at advanced stage of disease resulted in prolonged animal survival, decreased lymphadenosis and reduced levels of pathogenic or abnormal double negative CD4(-)CD8(-) cells and B220(+) cells in lymph nodes and spleen. We discovered that IIIM1 induces the production of an additional peptide, a fragment of alpha-1-antitrypsin, termed UBE. A relatively low dose (1 µg/kg) of UBE reduced proteinuria and hematuria in MRL/lpr mice. The beneficial effect of the peptide was corroborated by histological examination. Furthermore a significant reduction in serum IL17, IL12 and anti dsDNA antibodies was observed in the UBE-treated mice. Isolated CD4 cells incubated with the peptide showed a similar cytokine profile. Decreased levels of double negative CD4(-)CD8(-) and B220(+) cells were determined in lymph organs of UBE-treated animals. The beneficial effects of both UBE and IIIM1 suggest these peptides as potential drugs for SLE.

Amelioration of experimental autoimmune encephalitis by novel peptides: involvement of T regulatory cells.

The purpose of the present study was to develop a peptide for treatment of multiple sclerosis (MS). We have tested the effect of a novel anti-inflammatory peptide (KGHYAERVG, termed IIIM1) on experimental autoimmune encephalitis (EAE), an animal model of MS. Our findings demonstrate significant reduction in neurological score following oral administration of IIIM1. Structural studies revealed that the entire peptide is required for activity. The peptide caused significant reduction in IL17, interferon gamma, IL23 and IL12 production by isolated splenocytes and concomitant elevation of anti-inflammatory cytokines. IIIM1 elevated T regulatory cells (Tregs, CD4(+)CD25(+)FoxP3(+)) in brain and spleen of EAE mice. Similar proliferative effect was observed in isolated human and mouse Tregs in vitro. Stimulation of Tregs by IIIM1 caused production of a new peptide termed RA1 present in Oryza Sativa Japonica group. This Japanese rice peptide ameliorated neurological symptoms in the EAE model. Similar beneficial effect was observed upon oral administration of an extract of Japanese rice. In conclusion, oral treatment with IIIM1 ameliorates EAE symptoms via stimulation of Tregs to proliferate and produce RA1 which reduces EAE symptoms. RA1 might be involved in the relatively low prevalence of MS in Japan and other Japanese rice-eating populations.

Activity of histone H1.2 in infected burn wounds.

OBJECTIVES: Infections with multidrug-resistant microorganisms (e.g. Pseudomonas aeruginosa and Staphylococcus aureus) cause immense complications in wound care and in the treatment of immunosuppressed patients. Like most antimicrobial peptides, histones are relatively small polycationic proteins located in each eukaryotic nucleus, which naturally supercoil DNA. The aim of this study was to investigate the in vitro and in vivo activity of histone H1.2 in infected burn wounds and its potential toxicity. METHODS: To characterize the antimicrobial properties of histone H1.2 against potential causative organisms of burn wound infections, the in vitro radial diffusion assay and modified NCCLS microbroth dilution MIC assay were carried out. Haemolytic and cytotoxic properties were determined in human red blood cells and primary human keratinocytes. In vivo antimicrobial activity was tested in an infected rat burn model with P. aeruginosa (ATCC 27853). All results were compared with the naturally occurring broad-spectrum antimicrobial peptide protegrin-1 and with antibiotics clinically used against the corresponding bacteria. RESULTS: Human histone H1.2 exerted good antimicrobial activity against all tested microorganisms without significant haemolytic activity. Surprisingly, histone H1.2 showed cytotoxicity with an LD50 of 7.91 mg/L in primary human keratinocytes. The in vivo burn model data revealed a significant three-fold higher reduction in bacterial counts within 4 h compared with carrier control. CONCLUSIONS: These findings indicate that histone H1.2 is a potential candidate for use as a local and, because of its low haemolytic activity, systemic antimicrobial agent. However, further investigations are needed to specify the cytotoxicity and the dose-response relationship for histone H1.2.

Effect of electric charged molecules on Sindbis virus hemagglutination and hemolysis.

The role of electrostatic interactions in the attachment and fusion at acidic pH of Sindbis virus (SNV) with goose erythrocytes was studied, investigating the effect of several anionic and cationic polyelectrolytes on SNV hemagglutination and hemolysis. In order to establish the target of active drugs, the compounds were incubated either with the virus particles or with the erythrocytes. Dextran sulfate was the only compound able to inhibit the attachment of SNV to the erythrocytes. Fusion of virus with red cells was reduced dose-dependently by the polyanions dextran sulfate, mucin and polygalacturonic acid. On the contrary two polycations, polylysine and polybrene, enhanced viral hemolytic activity. However the effect of polyions is not exclusively related to the electric charge since ineffective molecules were found in both classes of compounds.

[Differentiation in vitro of chick-embryo skin in presence of histones. Analysis of protein synthesis in toto and of keratin in particular, with the use of labeled precursors]. / Studio del differenziamento in vitro della pelle di embrione di pollo in presenza di istoni. Analisi della sintesi proteica in toto e della cheratina in par ticolare con l'uso di precursori marcati.

Six-day limb skin from a chick embryo was cultured in vitro for seven days in a complete medium either supplemented or not with histones. At the end of the incubation period, the chick embryo skin cultured in the absence of histones was found to undergo keratinization, the converse being true for the limbs cultured in the presence of histones. In the latter, when H3-leucine and C14-cystine were added to the medium, a sharp decrease in the labeled amino acid incorporation was found.

[Further research on the differentiation in vitro of chick-embryo skin in the presence of histones]. / Ulteriori ricerche sul differenziamento in vitro della pelle di embrione di pollo in fresenza di istoni.

In the present study the authors have carried out further researches on the differentiation of six day limb skin from chick embryo cultured "in vitro" in a complete medium supplemented with histones. When histones were added to the medium in the first two days "in vitro", epidermal keratinization was not observed. However the addition of histones after four days "in vitro" did not interfere with epidermal differentiation.

[Comparative study of the antiviral properties of histones of animal and plant origin]. / Sravnitel'noe izuchenie antivirusnykh svoistv gistonov zhivotnogo i rastitel'nogo proiskhozhdeniia.

The antiviral properties of histones of animal (thymus) and plant (French beans) origin were studied in plants and with a plant virus, tobacco mosaic virus (TMV). Histones of the thymus and French beans were shown to be able to inhibit TMV reproduction. The antiviral properties of histones were found to depend on their concentration, pH, and to be determined by the modes of their introduction into leaves. The manifestation of the antiviral properties of histones seems to require not only their direct contact with virus but also a certain exposure on the leaf. The similarities of antiviral protective mechanisms of plants and animals determined by substances of the protein nature are discussed.

[Effect of novoimanine on the cellular permeability indices of staphylococci]. / Vliianie novoimanina na nekotorye pokazateli kletochnoi pronitsaemosti stafilokokkov.

Novoimanine is an antibacterial drug from Hypericum perforatum L. When used in the bacteriostatic concentration, i.e. 0.5 gamma/ml, it induced release of potassium ions from the cells of Staphylococcus aureus 209P and had no effect on release of the UV-absorbing compounds and 14C-amino acids. In addition, incubation of the cells with novoimanine (2.5--50 gamma/ml) provided "preservation" in them of the earlier absorbed 14C-amino acids, while in the control cells their level decreased. In a concentration of 100 gamma/ml novoimanine stimulated activity of ATP-ase and alkaline phosphatase by 34 and 37-57 per cent respectively. Histones F1 and F3 of the calf thymus induced an intensive release of 14C-amino acids from the cells of staphylococci and increased the activity of ATP-ase by 6-10 times. The data of the study suggested that the effect of novoimanine on the cytoplasmic membrane was limited and different from that on the polycationic antibacterial agents.

[Combination of novoimanine with antibiotics with a different mechanism of action]. / Sochetanie novoimanina s antibiotikami razlichnogo mekhanizma deistviia

Ampicillin, kanamycin, fusidic acid and rifocin significantly increased the effect of novoimanin on Staph. aureus 209. Histon F1 and spectinomycin did not influence the effect of novoimanin. Inefficiency of novoimanin combination with histon F1 provided a supposition that their effect may be directed to the same cell structures and most probably to the membranes. This was confirmed by the data of electron microscopy. The most effective combinations were recommended for the studies on their possible clinical use.