Early dynamics of glial fibrillary acidic protein and extracellular DNA in plasma of mice after closed head traumatic brain injury.

BACKGROUND: Glial fibrillary acidic protein (GFAP) in plasma is an established biomarker of traumatic brain injury (TBI) in humans. Plasma extracellular DNA (ecDNA) is a very sensitive, although nonspecific marker of tissue damage including TBI. Whether plasma GFAP or ecDNA could be used as an early non-invasive biomarker in the mouse model of closed head injury is unknown. The aim of this paper was to describe the early dynamics of plasma GFAP and ecDNA in the animal model of closed head TBI. METHODS: Closed head TBI was induced using the weight-drop method in 40 adult CD1 mice and blood was collected in different time points (1, 2 or 3h) after TBI in different groups of mice. Plasma GFAP and ecDNA and ecDNA fragmentation from the experimental groups were compared to healthy controls. In the surviving mice, a static rods test was performed 30 days after TBI to assess the neurological outcome of TBI. RESULTS: Despite a trend of higher plasma GFAP after TBI the differences between the groups were not statistically significant. Plasma ecDNA was higher by 50% after 1h (P<0.05) and 2h (P<0.05) after TBI and was highly variable after 3h. Plasma ecDNA, but not GFAP, was partially predictive of the neurological impairment of the mice. CONCLUSION: In this study, we have described the early dynamics of plasma GFAP and ecDNA after TBI in mice. According to our results, ecDNA in plasma is a more sensitive early marker of TBI than GFAP. Analysis of tissue-specific ecDNA might improve its predictive value regarding the survival and neurobehavioral outcome.

Hemispheric infiltrative grade II gliomas in adults: association of residual tumour volume and extent of tumour resection with malignant transformation.

OBJECTIVES: Evaluation of the impact of surgical treatment on malignant transformation (MT) of adult supratentorial infiltrative grade II gliomas (G2G) in a series of chemotherapy and radiotherapy-naïve patients. BACKGROUND: Despite G2G are slow-growing tumours, they typically undergo MT with a subsequent fatal disease course. An extensive resection alone likely changes their biological behaviour and defers MT; however, this impact is not unequivocally confirmed. METHODS: Thirty-eight chemotherapy and radiotherapy-naïve adult patients operated from 2005 till 2014 for a G2G were investigated. Based on postoperative magnetic resonance imaging (MRI) and/or positron emission tomography follow-up (FU) scans, the patients were classified as "transformers" (15 patients in whom MT occurred during the FU-period) and "non-transformers" (23 patients). RESULTS: The follow-up period of "non-transformers" was longer (p <0.0001). After adjustment for known risk factors - age, male sex, astrocytoma histology, preoperative tumour volume, preoperative contrast enhancement and positive isocitrate dehydrogenase 1 gene mutation status - a larger log postoperative tumour volume (p=0.031) and a smaller extent of resection (p=0.0086) were associated with a shorter MT-free survival. CONCLUSION: In our series, less extensive resections were associated with a shorter time to MT. Our data support an adoption of techniques enabling extensive G2G resections, such as intraoperative imaging and awake resections, into everyday routine (Tab. 1, Fig. 2, Ref. 40).

Sex Difference in Plasma Deoxyribonuclease Activity in Rats.

Extracellular DNA (ecDNA) activates immune cells and is involved in the pathogenesis of diseases associated with inflammation such as sepsis, rheumatoid arthritis or metabolic syndrome. DNA can be cleaved by deoxyribonucleases (DNases), some of which are secreted out of cells. The aim of this experiment was to describe plasma DNase activity in relation to extracellular DNA in adult rats, to analyse potential sex differences and to prove whether they are related to endogenous testosterone. Adult Lewis rats (n=28) of both sexes were included in the experiment. Male rats were gonadectomized or sham-operated and compared to intact female rats. Plasma ecDNA and DNase activity were measured using fluorometry and single radial enzyme diffusion assay, respectively. Concentrations of nuclear ecDNA and mitochondrial ecDNA were determined using real-time PCR. Females had 60% higher plasma DNase activity than males ( p=0.03). Gonadectomy did not affect plasma DNase in males. Neither the concentration of total ecDNA, nor nuclear or mitochondrial DNA in plasma differed between the groups. No significant correlations between DNase and ecDNA were found. From previous studies on mice, it was expected, that male rats will have higher DNase activity. In contrast, our study in rats showed the opposite sex difference. This sex difference seems not to be caused by endogenous testosterone. Interestingly, no sex differences were observed in plasma ecDNA suggesting a complex or missing association between plasma ecDNA and DNase. The observed sex difference in plasma DNase should be taken into account in animal models of ecDNA-associated diseases.

Acute effect of cola and caffeine on locomotor activity in drosophila and rat.

Caffeine is well known for reducing fatigue and its effect on behavior is widely studied. Usually, caffeine is not ingested in its pure form but rather in sugar-sweetened beverages such as cola. Our aim was to compare the acute effect of cola and caffeine on locomotor activity. Rats and flies ingested cola or caffeine solution for 24 hours. The open field test revealed higher locomotor activity in cola groups for both flies and rats. Surprisingly, no differences have been observed between caffeineand control group. We conclude that caffeine itself does not explain the effect of cola on locomotor activity. Effect of cola cannot be generalized and interpreted for any caffeinated drink with other contents. Rather, the observed effect on locomotor activity may be caused by interaction of caffeine with other substances present in cola.

The effect of long-term hypogonadism on body composition and morphometry of aged male Wistar rats.

Clinical studies show that hypogonadism in the aging male is associated with obesity and osteoporosis. Experimental studies are mostly conducted on relatively young adult animals and the induced hypogonadism lasts for a relatively short time. The present study aimed to describe the effect of long-term hypogonadism beginning in puberty on body composition, morphometry, and bone mineral density in aged male rats. Morphometric measurements and dual-energy X-ray absorptiometry were conducted at the age of 30 months on control and gonadectomized males. Long-term hypogonadism did not affect body weight, but led to a higher fat mass (by 26 %), lower lean mass (by 44 %), shorter body length (by 9 %), and anogenital distance (by 26 %), as well as to lower tail circumference (by 15 %) in comparison to control males. Lower bone mineral density (by 13 %) and bone mineral content (by 15 %) were observed in gonadectomized males. Results showing sarcopenic obesity and osteoporosis in this model of long-term hypogonadism might mimic the situation in aging males better than the widely used short-term hypogonadism induced in young animals. The morphometric analysis could potentially be a useful tool to study normal weight obesity without the need for specific equipment.

Measurement of Urea in the Saliva of Healthy Mice - a Pilot Study.

Salivary urea is studied as a non-invasive alternative for screening and monitoring of renal diseases. Its high variability prevents a wider clinical use. Animal experiments are needed to identify factors affecting this marker. The aim of this study was to describe the inter-individual variability of salivary urea in healthy mice, establish reference intervals, and analyse the effects of sex, age and body weight. Plasma and saliva samples were obtained from 37 male and 41 female healthy adult CD1 mice aged 13-69 weeks (body weight 22-51 g). The reference interval for salivary urea in heathy mice based on our results is 2.7-8.4 mmol/l (CV = 23 %). Multivariate analysis did not show any significant effect of age, sex, or body weight. In addition, salivary urea did not correlate with its plasma concentrations. The high variability of the promising salivary marker of kidney function in healthy mice requires further research before its use to diagnose or monitor renal failure in animal models of kidney diseases. Other potential confounders should be analysed, including intra-individual and pre-analytical variability. In addition, a normalization factor such as total salivary proteins or salivation rate is likely needed.

Testosterone and the brain: from cognition to autism.

Sex and gender matter in all aspects of life. Humans exhibit sexual dimorphism in anatomy, physiology, but also pathology. Many of the differences are due to sex chromosomes and, thus, genetics, other due to endocrine factors such as sex hormones, some are of social origin. Over the past decades, huge number of scientific studies have revealed striking sex differences of the human brain with remarkable behavioral and cognitive consequences. Prenatal and postnatal testosterone influence brain structures and functions, respectively. Cognitive sex differences include especially certain spatial and language tasks, but they also affect many other aspects of the neurotypical brain. Sex differences of the brain are also relevant for the pathogenesis of neuropsychiatric disorders such as autism spectrum disorders, which are much more prevalent in the male population. Structural dimorphism in the human brain was well-described, but recent controversies now question its importance. On the other hand, solid evidence exists regarding gender differences in several brain functions. This review tries to summarize the current understanding of the complexity of the effects of testosterone on brain with special focus on their role in the known sex differences in healthy individuals and people in the autism spectrum.

Does the 2nd and 4th digit ratio reflect prenatal androgen exposure?

OBJECTIVES:  The aim of our study was to describe the effect of prenatal testosterone exposure on 2D:4D in both sexes, and to determine whether this effect is mediated via the androgen receptor. In addition, the sex differences in lengths of 2D, 4D, and 2D:4D ratio were analyzed. BACKGROUND:  Clinical studies suggest a negative correlation between prenatal testosterone exposure and ratio of the lengths of the second and fourth digits (2D:4D). However, less is known about the underlying molecular mechanisms. METHODS:  Pregnant rats were treated with olive oil, testosterone, flutamide or testosterone with flutamide daily from the fourteenth day of pregnancy until delivery. The finger lengths of adult offspring were measured using both, digital scanning of the paws and µCT analysis of the phalanges. RESULTS:  None of the aforementioned methods revealed any effect of testosterone on 2D:4D. µCT measurements showed that prenatal hyperandrogenism in both sexes leads to shorter 2D compared to controls. Moreover, the testosterone treatment in males resulted in the shortening of 4D when compared to controls. CONCLUSION:  Prenatal hyperandrogenism leads to shorter lengths of 2D and 4D; however, it does not affect 2D:4D ratio. Whether other steroid hormones and/or testosterone metabolites affect the 2D:4D ratio requires further investigation (Tab. 5, Fig. 3, Ref. 32).

The role of sex hormones and social determinants in assessment of facial attractiveness.

OBJECTIVES: The current study is focused on assessing the factors of attractiveness of opposite-sex individuals based on evaluating photographs of their faces. BACKGROUND: When assessing the attractiveness, factors of both, the assessed individual and the assessor play a role. The relationship of the preference for partners based on their physical appearance with the markers of prenatal testosterone is not fully understood. METHODS: Sex hormone levels were measured in saliva, while age, social status, income and occupation were recorded. A total of 30 women and 35 men were enrolled. RESULTS: The identified factors determining the attractiveness of menare their age and prenatal testosterone level (second-to-fourth digit ratio - 2D:4D). The attractiveness of men is more influenced by the factors of evaluating women, namely the rating assigned to the men positively correlates with age, 2D:4D, and salivary estradiol of the evaluating women. The attractiveness of women correlated negatively with age and positively with prenatal exposure to androgens (2D:4D).The women with lower estradiol were rated higher by men who themselves had low estradiol levels. The attractiveness did not correlate with current testosterone. CONCLUSION: This study contributes to the knowledge on the role of sex hormones in human sexuality and partner choice. Further studies should include genetic factors of testosterone metabolism. (Tab. 4, Ref. 23).

Metabolic and renal effects of dietary advanced glycation end products in pregnant rats - a pilot study.

Thermally processed food contains advanced glycation end products (AGEs) including N(epsilon)-(carboxymethyl)lysine (CML). Higher AGEs or circulating CML were shown to be associated with pregnancy complications such as preeclampsia and gestational diabetes. It is unclear whether this association is causal. The aim of our study was to analyze the effects of dietary CML and CML-containing thermally processed food on metabolism in pregnant rats. Animals were fed with standard or with AGE-rich diet from gestation day 1. Third group received standard diet and CML via gavage. On gestation day 18, blood pressure was measured, urine and blood were collected and the oral glucose tolerance test was performed. Plasma AGEs were slightly higher in pregnant rats fed with the AGE-rich diet (p=0.09). A non-significant trend towards higher CML in plasma was found in the CML group (p=0.06). No significant differences between groups were revealed in glucose metabolism or markers of renal functions like proteinuria and creatinine clearance. In conclusion, this study does not support the hypothesis that dietary AGEs such as CML might induce harmful metabolic changes or contribute to the pathogenesis of pregnancy complications. The short duration of the rodent gestation warrants further studies analyzing long-term effects of AGEs/CML in preconception nutrition.

Markers of oxidative stress and antioxidant status in the plasma, urine and saliva of healthy mice.

Oxidative stress markers are usually measured in plasma, a stable environment for biomarkers. Blood collection is invasive, but the use of alternative biofluids is limited, due to high variability. In this study, we aimed to establish reference values for oxidative stress markers in plasma, urine and saliva of adult, healthy mice and to identify some sources of variability. Samples were obtained from 41 female and 37 male adult, healthy mice of the CD-1 strain, aged 95-480 days, weighing 21-55 grams. Reference ranges of TBARS (thiobarbituric acid reactive substances), AOPP (advanced oxidation protein products), fructosamine, GSH/GSSG (reduced and oxidized glutathione) ratio, TAC (total antioxidant capacity), and FRAP (ferric reducing antioxidant power) were measured in plasma and urine, and TBARS, GSH/GSSG ratio, TAC and FRAP in saliva, using standard spectrophotometric and fluorometric methods. Salivary GSH/GSSG and urinary AOPP were higher in females. Urinary fructosamine, GSH/GSSG and FRAP were higher in males. Urinary TAC and FRAP negatively correlated with age, and urinary GSH/GSSG positively correlated with weight. We determined that urine and saliva can be obtained non-invasively from mice, in sufficient amounts for reliable oxidative status assessment. Further studies are needed to uncover whether these biofluids reflect systemic oxidative status in diseases.

Sex differences in the effect of prenatal testosterone exposure on steroid hormone production in adult rats.

Maternal hyperandrogenism during pregnancy might have metabolic and endocrine consequences on the offspring as shown for the polycystic ovary syndrome. Despite numerous experiments, the impact of prenatal hyperandrogenic environment on postnatal sex steroid milieu is not yet clear. In this study, we investigated the effect of prenatal testosterone excess on postnatal concentrations of luteinizing hormone, corticosterone and steroid hormones including testosterone, pregnenolone, progesterone, estradiol and 7beta-hydroxy-epiandrosterone in the offspring of both sexes. Pregnant rats were injected daily with either testosterone propionate or vehicle from gestational day 14 until parturition. The hormones were evaluated in plasma of the adult offspring. As expected, females had lower testosterone and higher pregnenolone, progesterone and estradiol in comparison to males. In addition, corticosterone was higher in females than in males, and it was further elevated by prenatal testosterone treatment. In males, prenatal testosterone exposure resulted in higher 7beta-hydroxy-epiandrosterone in comparison to control group. None of the other analyzed hormones were affected by prenatal testosterone. In conclusion, our results did not show major effects on sex hormone production or luteinizing hormone release in adult rats resulting from testosterone excess during their fetal development. However, maternal hyperandrogenism seems to partially affect steroid biosynthesis in sex-specific manner.

How do we love? Romantic love style in men is related to lower testosterone levels.

Testosterone has been widely investigated in associations with many aspects of social interactions, emotions and behavior. No research has been conducted on its contribution to the variability of love styles in human. The aim of this paper was to uncover the possible relationship between not only the actual plasma testosterone levels, but also the prenatal testosterone level (expressed as 2D:4D ratio) and the sensitivity of androgen receptor and love typology in young healthy men. There are six love styles which are primary including Eros (passionate romantic love), Ludus (playful) and Storge (friendly) and secondary love consisting of Mania (obsessive), Pragma (practical realistic) and Agape (altruistic). Our results pointed out that low testosterone concentrations are associated with higher score for Eros, Ludus, Pragma, Mania love style. No significant association was proved for other tested parameters of androgenicity (2D:4D, sensitivity of androgen receptor) and love style after correction was applied. Different attitudes and behavior in relationships do have a biological foundation related to endogenous testosterone levels in plasma. Future studies should address questions about the family and social background of participants to differentiate here between moral rules or/and social-conventional rules.

Beneficial effects of live and dead Salmonella-based vector strain on the course of colitis in mice.

Dysbiosis of intestinal microbiota and hyperactive immune responses seem to be crucial for the uncontrolled inflammation in inflammatory bowel diseases (IBD). Modulation of the microbiome and immune stimulation of the intestinal epithelium were suggested as therapeutic approaches. In this study, live attenuated and dead bacterial cells of Salmonella Typhimurium SL7207 - a widely used bacterial vector for gene therapy were administered in DSS-induced colitis in mice. C57BL/6 mice were divided into four groups. The first group received pure water (CTRL). The other three groups received 2% dextran sulphate sodium (DSS) to induce colitis. Two DSS groups were treated with live attenuated (DSS live) or inactivated (DSS dead) Salmonella by gastric gavage. Intake of 2% DSS caused weight loss in all DSS groups compared to control mice with some improvement in DSS live group on the last day of the experiment. Significantly longer colon and improved stool consistency were reported in DSS live group, but not DSS dead group, when compared with DSS. Significant enlargement of spleens was observed only in DSS and DSS dead groups compared to control. Significant differences in stool consistency, colon length and spleen enlargement were observed between DSS live and DSS dead groups with beneficial effects of live bacteria. Interestingly, significant decrease in myeloperoxidase activity was detected in both, DSS live and DSS dead groups compared to the DSS group. On the basis of these results, progression of colitis seems to be beneficially influenced not only by live attenuated but to some extent also by inactivated Salmonella Typhimurium SL7207. Our results provide evidence that Salmonella-based gene therapy vectors are able to positively alter gut homeostasis during DSS-induced colitis. SIGNIFICANCE AND IMPACT OF THE STUDY: Restoration of gut homeostasis has a great importance in IBD. Here, we tested the nonspecific effect of the strain Salmonella Typhimurium SL7207 on the course of colitis to find out whether the potential effect would be mediated by activity of live bacterial cells or by bacterial structures that are also present in dead bacteria. Live bacterial therapy of colitis showed a beneficial effect on clinical signs as well as on macroscopic and inflammatory markers of colitis. On the other hand, therapy with dead bacteria showed inconsistent effects, negative in most clinical outcomes, positive especially in myeloperoxidase activity. Our data indicate that the beneficial effect of bacterial gene therapy vectors carrying therapeutic genes might be, at least partially, caused by the bacterial vector instead of the therapeutic gene.

Anthocyanin-Rich Diet in Chemically Induced Colitis in Mice.

The aetiology of inflammatory bowel diseases is unclear, but oxidative stress plays a key role in the pathogenesis. Anthocyanins--plant polyphenols--were shown to have antioxidant and anti-inflammatory properties. The aim of this study was to investigate the potential protective effects of anthocyanins on the oxidative status in mice with chemically induced colitis. Adult male mice were randomly divided into a control group drinking tap water and a colitis group drinking 1% dextran sulphate sodium solution. Animals had ad libitum access to a control wheat-based diet or food based on wheat producing anthocyanins. Bodyweight and stool consistency were monitored daily for 14 days. At the end of the experiment, colon length was measured and tissue samples were collected for the assessment of histology and oxidative status. Mice with colitis had lower body weight, higher stool score and shorter colon than control mice. Anthocyanins had neither an effect on stool consistency, nor on bodyweight loss and colon length. In the colon, liver and plasma, analysis of oxidative stress markers and antioxidant status revealed no significant differences between the groups. Food made from wheat producing anthocyanins did not protect mice from the consequences of chemically induced colitis. The measured biomarkers do not confirm the role of oxidative stress in this model of colitis. Further optimization of the anthocyanin-rich food might be needed before further experiments are conducted.

Does rat fetal DNA induce preeclampsia in pregnant rats?

Cell-free fetal DNA in maternal circulation is higher during preeclampsia. It is unclear whether it is the cause or the consequence of the disease. The aim of this study was to prove whether injected rat fetal DNA induces preeclampsia-like symptoms in pregnant Wistar rats. They received daily i.p. injections of water or rat fetal DNA (400 µg) from gestation day 14 to 18. Blood pressure, proteinuria, placental and fetal weight were measured at gestation day 19. Plasma DNase activity, proteinuria and creatinine clearance were assessed. There was no significant difference in any of the measured parameters. The results of this study do not confirm the hypothesis that fetal DNA might induce preeclampsia. This is in contrast to others using human fetal DNA in mice. Further studies should be focused on the effects of fetal DNA from the same species protected from DNase activity.

Doxorubicin-induced behavioral disturbances in rats: protective effect of melatonin and captopril.

OBJECTIVE: Doxorubicin is a recognized chemotherapeutic agent widely employed in human malignancies. The limiting factor of its use is a number of side effects. The aim of this work was to show, whether administration of doxorubicin could induce behavioral disturbances in rats, and whether angiotensin-converting enzyme inhibitor captopril or melatonin can modify these potential alterations. DESIGN AND METHODS: Four groups of 3-month-old Wistar rats (twelve per group) were treated for 4 weeks: control (placebo-treated), doxorubicin (DOX) (5mg/kg i.v. in a single intravenous dose), DOX rats treated with either melatonin (10mg/kg/24h) or captopril (100mg/kg/24h). Systolic blood pressure (SBP) and the level of oxidative stress were investigated and behavioral tests of anxiety-open field test (OF), elevated plus maze (EPM) and light-dark box (LDB) were accomplished. RESULTS: Doxorubicin increased significantly systolic blood pressure and parameters of oxidative stress. Moreover, doxorubicin enhanced the level of anxiety in the tests of OF, EPM, and LDB. Captopril and melatonin prevented the blood pressure rise and the enhancement of oxidative load. Importantly, both substances reduced the parameters of anxiety. CONCLUSION: Chronic administration of captopril or melatonin has shown anxiolytic effect in the model of doxorubicin-induced anxiety. It does not seem unreasonable to suppose that this protective effect of captopril or melatonin against anxiety development might have been related to the antioxidative effects of both substances.

Testosterone and estradiol are not affected in male and female patients with obstructive sleep apnea treated with continuous positive airway pressure.

BACKGROUND: Continuous positive airway pressure (CPAP) improves symptoms in patients with obstructive sleep apnea syndrome (OSAS). It is currently unclear, whether CPAP also alters endocrine parameters such as sex hormone levels. In a previous study, we have found no changes in sex hormones in patients with OSAS after one night with CPAP. AIM: The aim of this study was to prove long-term effects of CPAP on sex hormone concentrations in patients with OSAS. METHODS: Twenty-two women and 67 men with severe OSAS (respiratory distress index > 30/h) were enrolled in the study. Fasting blood venous samples were taken before CPAP therapy and after 1 and 6 months of CPAP treatment. Testosterone and estradiol were measured in all samples using commercially available ELISA kits. RESULTS: No effects of long-term CPAP treatment were found on testosterone or estradiol levels in OSAS patients of either gender. CONCLUSIONS: The results are in line with previous smaller studies. However, our study is larger and longer than previously published studies. In addition, this is the first study analyzing the effects of CPAP on testosterone and estradiol and in both genders. Positive effects of CPAP on sexual functions reported in other studies might, thus, be mediated by other than endocrine effects.

Reference values of skin autofluorescence as an estimation of tissue accumulation of advanced glycation end products in a general Slovak population.

AIMS: For decades, Slovakia has maintained a prominent place in mortality rates from cardiovascular diseases among European Union (EU-27) countries. Determination of skin autofluorescence serves as an estimate of tissue accumulation of advanced glycation end products--substances accumulating in tissues and body fluids that play a pathophysiological role in age-related diseases and their complications, such as diabetes. METHODS: In 1385 apparently healthy Slovakian subjects aged from a few days old to 77 years, skin autofluorescence was determined using an advanced glycation end product reader and compared with reference data from Dutch Caucasians. The impact of the weekly frequency of recreational physical exercise on skin autofluorescence was investigated in the adults, and the impact of feeding regimen in the infants. RESULTS: With the exception of 10- to 19-year-olds, Slovaks had lower skin autofluorescence values in comparison with the Dutch Caucasians. In healthy non-smokers, physical exercise for > 30 min/day performed ≥ 3 times/week was associated with lower skin autofluorescence levels. In infants, breastfeeding (advanced glycation end product-poor diet) was associated with lower skin autofluorescence levels in comparison with consumption of infant formulas (advanced glycation end product-rich diet). CONCLUSIONS: Reference ranges of skin autofluorescence in Slovak Caucasians, detailed for paediatric age groups, are provided. Our data show that, in healthy adults, regular physical exercise associates with lower skin autofluorescence. Infants fed or weaned from infant formulas (advanced glycation end product-rich diet) have higher skin autofluorescence than their breast milk-consuming counterparts. It is unclear why Slovaks have lower skin autofluorescence compared with a Dutch population with lower cardiovascular mortality rates. Reference data on skin autofluorescence from diverse populations are needed for the precise clinical interpretation of skin autofluorescence measurements.

On the origin and diagnostic use of salivary RNA.

Saliva as a diagnostic fluid enables non-invasive sampling, which can be performed even by an untrained person. Saliva is, thus, particularly useful for large population screenings, for children, elderly and whenever repeated samplings are needed. Saliva is a plasma filtrate actively modified by the salivary glands. Saliva could replace some routine blood tests in the future. The sources of salivary RNA include oral epithelial cells and oral micro-organisms. Recent developments suggest that using known salivary RNA markers, it is possible to diagnose diseases such as oral carcinoma and other diseases will be added soon. Salivary RNA can be used to identify oral bacteria and to determine the expression of specific genes. On a systemic level, it provides information about the whole oral transcriptome and microbiome. Despite the small amount of salivary RNA, the issues with its isolation have been overcome. Saliva, thus, contains RNA of sufficient quality and quantity for sensitive and specific analyses. Salivary RNA can provide medically relevant information about oral microbiome, oral carcinoma, but also breast and pancreatic cancer and is, thus, a promising tool for future research and clinical diagnostics.