Reduction of SARS-CoV-2 viral load in exhaled air by antiseptic chewing gum: a pilot trial.

PURPOSE: The dominant route of transmission of SARS-CoV-2 is airborne, through respiratory transmission by aerosols or droplets which can be measured by viral load in exhaled air. Several natural substances have shown antiviral activity. The aim of this pilot study was to investigate the effect of a chewing gum containing natural antiseptic ingredients (cinnamon-, peppermint- and lemon-oil, quercetin, spermidine, ginger and ginseng) on viral load in exhalative air in patients infected with SARS-CoV-2. METHODS: Nine patients infected with SARS-CoV-2 were enrolled and exhaled forcefully into a special mouthpiece at different time points before and after chewing the antiseptic gum. The mouthpiece contained a filter paper serving for extraction of coronaviruses following real-time PCR to quantify the viral load. RESULTS AND CONCLUSION: Cycle threshold (Ct) values of all patients increased after chewing the gum. The mean difference between the Ct values at baseline (before chewing the antiseptic gum) and time point 30 min (15 min after chewing) was 3.8 ± 2.6; (93% viral load reduction; p = 0.002). Time point 15 min (2.7 ± 1.7 (83% viral load reduction; p = 0.003)), 60 min (3.0 ± 3.4 (88% viral load reduction; p = 0.028)), 90 min (3.7 ± 1.8 (92% viral load reduction; p = 0.004)) and 120 min (3.0 ± 3.7 (91% viral load reduction; p = 0.05)) showed similar results. The antiseptic chewing gum demonstrated a significant potential to reduce SARS-CoV-2 viral load in exhalative air and, in this way, reduce further spread and infection risk. Larger placebo-controlled clinical trials are required to confirm these findings further.

Selenium in blood, semen, seminal plasma and spermatozoa of stallions and its relationship to sperm quality.

The essential trace element selenium is indispensable for male fertility in mammals. Until now, little data existed regarding the relationship between selenium and sperm quality in the stallion. Selenium, or selenium-dependent glutathione peroxidase activity, was determined in red blood cells, semen, seminal plasma and spermatozoa, and the percentages of spermatozoa with progressive motility (PMS), intact membranes (PMI), altered (positive) acrosomal status (PAS) and detectable DNA damage, determined by the sperm chromatin structure assay, were evaluated in 41 healthy stallions (three samples each). The pregnancy rate per oestrus cycle (PRC) served as an estimation of fertility. An adverse effect on stallion fertility caused by low dietary selenium intake was excluded, as all stallions had sufficient selenium levels in their blood. Interestingly, no significant correlations (P > 0.05) between the selenium level in blood and the selenium level in seminal plasma or spermatozoa were found, suggesting that the selenium level in blood is no indicator of an adequate selenium supply for spermatogenesis. The selenium level in spermatozoa (nmol billion(-1)) was correlated with PMI, PMS and PAS (r = 0.40, r = 0.31 and r = -0.42, respectively; P

Metal-containing proteins in the apoptosis and redox processes in the rat prostate and human prostate cells.

Several trace elements, such as Se, Cu, Mn, and Zn are bound to proteins (metallo- and metalloidproteins) in the prostate gland. Currently, it is known that some of those elements play a role in the apoptosis of different cells and redox processes. For the detection of such proteins, analytical and biochemical procedures were combined. SEC and ICP-MS were used to detect some trace elements, which are bound to proteins in the prostate cytosol and/or in the human prostate cell lines. Several antibodies against specific proteins were tested. By means of some of these antibodies several trace element-containing proteins, such as selenoproteins and Cu- and Cu-Zn-proteins, could be identified in the prostate. In addition, the localization of such metal- and metalloid-containing proteins in the micro organelles and cytosol of the prostate indicates specific functions of these proteins because, as it is known, such metal- and metalloid proteins play a role in the apoptosis and especially in the redox processes.

Herpesvirus infection accelerates atherosclerosis in the apolipoprotein E-deficient mouse.

BACKGROUND: Human herpesviruses have been implicated but not proven to be involved in the etiology of atherosclerosis. To determine whether there is a causal relationship, the effect of herpesvirus infection on the development of atherosclerosis was assessed in the apolipoprotein E-deficient (apoE-/-) mouse. METHODS AND RESULTS: In the present study, 3- to 4-week-old apoE-/- mice were infected with murine gamma-herpesvirus-68 (MHV-68). Atheroma formation was accelerated over a 24-week period in infected apoE-/- mice compared with control uninfected apoE-/- mice. Acceleration of atherosclerosis was reduced by antiviral drug administration. Histological analysis of the atheromatous plaques showed no difference between lesions of infected and control mice. Viral mRNA was present in the aortas of infected mice before lesion development on day 5 after infection. This suggests that the virus may initiate endothelial injury, which is believed to be an early event in the development of atherosclerosis. Therefore, the virus may play a direct role in atherosclerosis rather than be an "innocent bystander." CONCLUSIONS: These data demonstrate that a gamma-herpesvirus can accelerate atherosclerosis in the apoE-/- mouse. This study provides the first report of a murine model in which to study the causative role of herpesvirus infection in the development of atherosclerosis.

Association between Kaposi's sarcoma and atherosclerosis: implications for gammaherpesviruses and vascular disease.

The treatment of HIV-positive patients with protease inhibitors has been suggested to increase their risk of atherosclerosis. The cause of this accelerated atherogenesis is unknown, but on the basis of previous studies we postulated that it could be linked to the presence of human herpesvirus-8. A retrospective analysis of post-mortem reports showed a strong correlation between Kaposi's sarcoma and the presence of atheroma. This hypothesis merits further investigation.

Partial inhibition of vesicular stomatitis virus by the interferon-induced human 9-27 protein.

To determine whether the interferon-induced 9-27 protein of human cells contributes to the antiviral state, we expressed the 9-27 cDNA under the control of a constitutive promoter and assayed transfected cells for enhanced virus resistance. The intracellular distribution of 9-27 resembled that of cytoskeleton-associated proteins. Analysis at the single-cell level by indirect immunofluorescence revealed that mouse cells expressing 9-27 were less permissive for vesicular stomatitis virus than control cells not expressing 9-27. No significant inhibition of influenza virus was observed. When tested in parallel, 9-27 was found to have less powerful antiviral activity toward vesicular stomatitis virus than the interferon-induced MxA protein. Thus, 9-27 joins the family of interferon-induced proteins with intrinsic antiviral activity.

The production of a truncated form of baculovirus expressed EHV-1 glycoprotein C and its role in protection of C3H (H-2Kk) mice against virus challenge.

A truncated form of the equine herpesvirus 1 (EHV-1) glycoprotein C (gC) gene was expressed in baculovirus. The gC signal sequence was substituted with the honeybee melittin signal sequence and the transmembrane region was replaced with a histidine tag. The recombinant virus produced high levels of gC in both the cells and supernatants of infected cells. The protein was present by 24 h and maximal secretion occurred at 96 h post-infection. The recombinant protein was antigenically authentic as shown by its reaction with each of a panel of individual monoclonal antibodies specific for the five distinct antigenic sites on EHV-1 gC. Recombinant gC was purified from the supernatant of infected cells by immuno-affinity chromatography and used to immunize C3H (H-2Kk haplotype) mice. This incurred a gC specific antibody response against both the recombinant protein and EHV-1 gC. 'Pepscan' analysis showed that the gC specific antibodies in serum from these mice reacted with the same epitopes on gC as those recognized by antibodies in convalescent equine sera (i.e. antibodies were specific to antigenic sites one and five). A third previously unrecognized antibody binding site at the carboxyl terminus was also detected (Antibody binding domain I). A T-cell proliferative response against EHV-1 was detected in splenocyte populations taken from vaccinated mice. Further, the recovery of virus from the lungs and turbinates following challenge of mice with EHV-1 was significantly reduced. These findings indicate that baculovirus expressed gC may contribute significantly to a subunit vaccine preparation aimed at protecting horses from EHV-1 infection.

The expression of the proteins of equine herpesvirus 1 which share homology with herpes simplex virus 1 glycoproteins H and L.

Several expression systems were used in studies aimed at characterizing the equine herpesvirus 1 (EHV-1) glycoprotein H and L homologues of HSV-1 (EHV-1 gH and gL) and the products were compared to the authentic proteins synthesized in virus infected cells. Using an in vitro transcription/translation system two gH species were detected (an unprocessed 89 kDa and a processed 116 kDa product). Three low molecular weight proteins were found in the case of gL (21.8 kDa, 22.9 kDa and 26.9 kDa) and these showed a slight reduction in mobility on the addition of microsomal membranes to the reactions. A gL fusion protein was produced in pGEX-2T, expression being confirmed by Western blotting using a gL-specific antiserum raised against a peptide incorporating the 13 carboxyl terminal amino acids of the protein. A gH specific peptide antiserum precipitated both gH and two smaller proteins from EHV-1 infected cells thought to be two forms of gL. Insect cells infected with gH or gL baculovirus recombinants were used to vaccinate C3H (H-2k) mice. Some protection against EHV-1 infection was conferred to the gH inoculated mice. The results will enable further studies on the importance of the gH and gL interaction in the pathogenesis of EHV-1 to be evaluated and their potential in contributing to a subunit vaccine to be assessed.

Profiles of eicosanoid production from 14C-arachidonic acid and prostaglandin metabolism by rabbit esophageal mucosa and muscularis.

In an attempt to elucidate the possible involvements of eicosanoids in esophageal functions and disorders, we have investigated the formation of both cyclooxygenase and lipoxygenase metabolites from 14C-arachidonic acid by rabbit esophageal tissues. Homogenates of rabbit esophageal mucosa and muscularis were incubated with 14C-arachidonic acid and after ether extraction eicosanoids were separated and quantified by reverse phase high performance liquid chromatography. The predominant cyclooxygenase products were 6-keto-PGF1 alpha, PGF2 alpha, and PGE2 for mucosa and 6-keto-PGF1 alpha, and PGE2 for muscularis. The formation of these products was inhibited both by indomethacin and the dual pathway inhibitor, nordihydrogualaretic acid (NDGA). In mucosa the major eicosanoid was 12-HETE (12-hydroxyeicosatetraenoic acid) which was inhibited by NDGA but not by indomethacin which on the contrary enhanced its formation. Additionally four polar products were synthesized which appeared to be lipoxygenase-dependent as their formation was inhibited by NDGA but not by indomethacin. Muscularis produced as a minor lipoxygenase product only 12-HETE, which was inhibited by NDGA but unchanged in the presence of indomethacin. In addition, both tissues, but mucosa more than muscularis, possessed large prostaglandin catabolizing capacity. The present findings indicate that rabbit esophageal tissues can convert 14C-arachidonic acid into lipoxygenase as well cyclo-oxygenase products which may have a role in esophageal physiology and pathophysiology.

Effect of chronic REM sleep deprivation on pituitary, hypothalamus and hippocampus PGE2 and PGD2 biosynthesis in the mouse.

Given the often reported relationships between sleep-wake regulation and the cerebral prostaglandins (PGs), the effect of chronic rapid eye movement (REM) sleep deprivation on brain PGE2 and PGD2 biosynthesis in mouse was evaluated, since they are known to have opposite actions as respectively wake- and sleep-inducing substances. Mice were subjected to 5 and 10 days of REM sleep deprivation by the flower pot technique. After sacrifice, PGE2 and PGD2 were determined in the pituitary, hypothalamus and hippocampus. Except in the pituitary where no changes were shown, the PGE2/PGD2 ratio was significantly enhanced after 5 and 10 days of REM sleep loss, when compared to control. These results showed an alteration of cerebral PGE2 and PGD2 biosynthesis, resulting in a shift from PGD2 toward PGE2. These results were not consistent with a role of PGD2 as a sleep-promoting substance as, if that was the case, it would be increased during the REM sleep deprivation. But they do not rule out its involvement as a facilitating substance.

In vivo and in vitro magnesium effects on aortic prostacyclin generation in DOCA-salt hypertensive rats.

To investigate the blood pressure lowering effect of magnesium (Mg2+) in the hypertensive rat, we measured the prostacyclin release (PGI2, as immunoreactive 6-keto-PGF1 alpha) by isolated aortae from normotensive and deoxycorticosterone acetate (DOCA)-salt hypertensive rats fed a control or Mg(2+)-enriched diet. We also studied the in vitro effect of Mg2+ on aortic PGI2 release. The Mg(2+)-enriched diet significantly decreased by 10% blood pressure in DOCA-salt hypertensive rats but not in normotensive rats. The Mg(2+)-enriched diet significantly increased by 122% aortic PGI2 release in DOCA-salt hypertensive rats, but not in normotensive rats. Mg2+ supplementation in the incubation medium (4.8 mM) significantly increased aortic PGI2 release by 94% in DOCA-salt hypertensive rats, but not in normotensive rats. These data suggest that the Mg(2+)-induced attenuation of blood pressure in DOCA-salt hypertensive rats could be linked with the enhanced vascular PGI2 release.

A drug used in traditional medicine, harpagophytum procumbens: no evidence for NSAID-like effect on whole blood eicosanoid production in human.

Devil's Claw (Harpagophytum procumbens), an herbal product being marketed in Canada and in Europe as a home remedy for the relief of arthritic disease, was investigated in healthy humans on eicosanoid production during spontaneously blood clotting. Volunteers took H. procumbens (daily 4 capsules of 500 mg powder containing 3% of total glucoiridoids) for a period of 21 days. The following are the results (mean (SEM)): before H. procumbens intake, prostaglandin (PG)E2 (ng/ml serum): 2.1 (0.4) (n = 25), thromboxane (TX)B2: 147 (27) (n = 25), 6-keto-PGF1 alpha: 4.4 (0.7) (n = 13), leukotriene (LT)B4: 3.4 (0.4) (n = 25); after intake: PGE2: 3.2 (0.6), TXB2: 143 (24), 6-keto-PGF1 alpha: 4.2 (0.9), LTB4: 3.8 (0.6). Each subject serving as her own control, no statistically significant differences were observed between before and after H. procumbens intake. These results indicate that Devil's Claw lacks, at least in healthy humans and under the selected conditions, the biochemical effects on arachidonic acid metabolism of antiarthritic drugs of the non-steroidal antiinflammatory type.

Predictive value of urinary N-acetyl-beta-D-glucosaminidase (NAG), alanine-aminopeptidase (AAP) and beta-2-microglobulin (beta 2M) in evaluating nephrotoxicity of gentamicin.

Concentrations of N-acetyl-beta-D-glucosaminidase (NAG), alanine-amino-peptidase (AAP) and beta-2-microglobulin (beta 2M) were determined daily in the urine of 28 patients treated with gentamicin (2-3 mg . kg-1 . day-1) for a mean of 15 days. All had normal renal function. Increased activity in NAG and AAP was observed for all patients, either immediately or after 2 or 3 days of treatment. The results were compared with serum creatinine concentrations and urinary beta 2M levels. This study indicates a relationship between the nephrotoxicity of gentamicin and initial urinary enzymic activity(NAGi) prior to any treatment. The degree of NAG response during the first ten days of treatment appeared as a second prognostic factor. Renal failure was observed for one out of the 12 patients with normal NAGi (NAGi less than 200 mumol/day). Seven of them showed a marked enzyme activity response (greater than 1500 mumol/day) with an increase in beta 2M activity. Eleven out of the 16 patients with elevated NAGi (NAGi greater than 200 mumol/day) developed renal failure and showed an elevated maximal response. The concentration of AAP appears to be of little prognostic value. The variation in individual maximal urinary enzyme responses observed among the 28 patients during the first ten days of treatment points to the existence of individual sensitivities to gentamicin, the exact mechanism of which remains unclear.

Role of T-cells, virus neutralising antibodies and complement-mediated antibody lysis in the immune response against equine herpesvirus type-1 (EHV-1) infection of C3H (H-2k) and BALB/c (H-2d) mice.

The suitability of C3H (H-2k) and BALB/c (H-2d) mice for use as small animal models to study immunity to EHV-1 was assessed. An in vitro T cell response mediated by both CD4+ and CD8+ T cells was detected both during the acute phase of infection and after challenge with a second dose of EHV-1 at two months in lymphocyte populations taken from the spleens of both types of mouse. The responses were apparent until at least 61 days after the primary inoculation. After challenge, T cells from mice previously infected with EHV-1 responded by as early as day 3 after infection and higher levels of T cell proliferation were reached than in mice undergoing a primary infection. Immunological cross-reactivity with the closely related virus, EHV-4 was detected and some activity against herpes simplex virus type-1 (HSV-1) was observed during the acute phase of infection. T cell responses were detected in the draining cervical lymph nodes but not in the inguinal lymph nodes of the mice and these were the primary sites of T cell activation. Complement-dependent virus neutralising antibodies were present by day 8 after infection. These antibodies were also able to lyse EHV-1 infected target cells in vitro. Complement-independent virus neutralising antibodies were found before challenge only in C3H mice. The clinical signs and duration of virus shedding were reduced after challenge. The time course of the appearance of the different immune effector mechanisms is discussed in relation to the clearance of virus from the infected mice. The results suggest that C3H mice provide a better model in which to study potential vaccine candidates against EHV-1 infections of the horse than BALB/c mice.

Determination of the beta- branching ratio of 64Cu by mass spectrometric investigations of the decay products in neutron transmuted copper.

The beta- branching ratio of 64Cu was determined by investigating the resulting decay products in copper doped by neutron transmutation. The numbers of 64Zn and 64Ni atoms were analyzed using isotope dilution analysis combined with thermal ionization mass spectrometry. A beta- branching ratio of (38.06+/-0.30)% was obtained, which agrees with the study of Kawada (Appl. Radiat. Isot. 37 (1) (1986) 7) to a higher accuracy. However, our result differs from the value cited in the NUDAT database of (39.0+/-0.3)%.

Term and preterm labour are associated with distinct microbial community structures in placental membranes which are independent of mode of delivery.

Infection is considered a possible trigger for preterm labour, supported by evidence showing the presence of bacteria in the placenta and placental membranes from preterm births. In this study, 16S rDNA pyrosequencing was used to identify bacteria in placental membranes. Caesarean sections and vaginal deliveries at term were found to harbour common genera. Mycoplasma hominis, Aerococcus christensenii, Gardnerella vaginalis and Fusobacterium nucleatum were either only present in preterm membranes or in greater abundance than at term. These data support previous studies that used either targeted qPCR or broad-range 16S rDNA PCR and cloning but not a recent microbiome analysis of placental tissue using high-throughput sequencing.

Effect of a swim training on homocysteine and cysteine levels in rats.

The purpose of this study was to investigate the effects of a 8-week of swim training on total plasma homocysteine and cysteine levels in 16 male Sprague-Dawley rats aged 17 weeks. We also evaluated the activity of hepatic cystathionine beta-synthase (CBS), an enzyme involved in the metabolism of Hcy, the concentration of plasma glutathione, taurine, and a fraction of vitamin B6: the pyridoxal 5-phosphate (PLP). After one week of acclimatization, rats were randomly divided into two groups: 8 non-trained (NTR) and 8 trained rats (TR). Following the training period, body weight gain was lower in TR than in NTR. Plasma homocysteine did not differ among groups while significantly lower plasma cysteine and taurine levels were found in TR (157.83 +/- 8.6 micromol/L; 133.01 +/- 9.32 micromol/L; P < 0.05) compared with data of NTR (176.19 +/- 4.9 micromol/L; 162.57 +/- 8.16 micromol/L; P < 0.05). No significant changes in hepatic CBS activity were observed in TR compared with NTR. Moreover, values for plasma glutathione and PLP concentrations were not affected by training.These results indicate that training reduces plasma cysteine and taurine levels whereas it does not modify other studied parameters. Thus, physical training may regulate cysteine metabolism.

Determination of Zn in high-purity GaAs with neutron activation analysis

Neutron activation analysis was used to analyse Zn concentrations in high-purity GaAs. A combination of chemical separation of the neutron-induced 65Zn from the GaAs-matrix with anion-exchange chromatography and ultra low-level gamma-ray spectrometry in an underground laboratory was applied. Zn concentration ranged from 2.1 to 51 ng g(-1), and detection limits from 0.018 to 0.059 ng g(-1) were obtained.