Salivary Enzymes Research Articles

Salivary alpha-amylase enzyme is a non-invasive biomarker of acute stress in Japanese macaques (Macaca fuscata).

Salivary alpha-amylase (sAA) enzyme functions as a digestive enzyme in many species that consume starch in their diet. Human studies have also revealed that sAA enzyme activity levels are positively correlated with the release of the stress hormone norepinephrine, allowing sAA to act as a biomarker for sympathetic nervous system activity. Recent non-human primate studies have incorporated sAA as a physiological stress marker. However, no published reports have investigated the time course of sAA from a stressful event to return to baseline levels in non-human primates. Furthermore, no validation of sAA as a stress biomarker has been reported for Japanese macaques (Macaca fuscata). This study had two primary aims: (1) to develop a systematic method for non-invasive saliva collection and, (2) to investigate sAA as a biomarker of acute stress in M. fuscata in order to better understand its acute stress-related characteristics. We developed a non-invasive method for cooperative saliva collection using positive reinforcement training (PRT) and tracked individual progress over 595 trials in ten individually housed Japanese macaques. We detected sAA enzyme in M. fuscata via kinetic reaction assay, then performed 22 acute stress tests. Four tests met conditions for interpreting sAA in response to an acute stressor and these results show that on average sAA activity rapidly increased post-stressor (mean ± SD = 4.2 ± 0.9min) and returned to baseline shortly thereafter (10.4 ± 0.6min). Our report reveals for the first time the temporal dynamics of sAA when applying acute stress to Japanese macaques and could be a useful tool for assessing animal welfare.

Efficacy of Brazilian Propolis Supplementation for Japanese Lactating Women for Atopic Sensitization and Nonspecific Symptoms in Their Offspring: A Randomized, Double-Blind, Placebo-Controlled Trial.

Propolis is a natural product collected from several plants by honeybees and mixed with beeswax and salivary enzymes. In animal models, propolis suppressed IgE-mediated allergies. However, there is no clinical evidence that propolis prevents human atopic sensitization, to the best of our knowledge. Therefore, a randomized, double-blind, placebo-controlled trial was conducted to assess whether propolis supplementation for lactating women increases or decreases the level of total IgE and antigen-specific IgE in the serum of their offspring (i.e., atopic sensitization) at the time of their first birthday. In addition, whether propolis supplementation improves or worsens nonspecific symptoms (e.g., eczema) in the lactating women and their offspring was also investigated. This trial is registered with UMIN000020794. Eligible pairs of mothers and their offspring (n=80) were randomized to two groups: propolis (n=40) and placebo (n=40). Participants were evaluated every month, and 31 (78%) of the propolis group and 23 (58%) of the placebo group underwent blood tests at the first birthday of the offspring. Total IgE ≥ 10 UA/ml was seen in 26 (84%) infants whose mothers were given propolis, which was not significantly different from the 19 (86%) given placebo (P=0.80). Total IgE as a continuous variable was not significantly different between the propolis and placebo groups (P=0.70). Antigen-specific IgE levels for mites, egg white, cow's milk, and wheat, as both dichotomous and continuous variables, were not significantly different between the two groups. Both in mothers and their offspring, there were no significant differences in the subjective improvements of nonspecific symptoms between the two groups. Except for one mother who had transient and mild nausea, none of the other mothers or their offspring developed severe adverse events during the follow-up period. In conclusion, compared with placebo, Brazilian propolis supplementation did not influence the risk of atopic sensitization in infants and neither did it improve nor worsen nonspecific symptoms in either mothers or their infants.

The Physiology and Biochemistry of the Digestion System of Termites from the Genus Anacanthotermes Jacobson, 1904

A variety of morphological and functional features of the digestive system of termites is associated with their nutritional adaptation. Wood is mostly the food of termites’ adult larvae, workers and young nymphs. The salivary and intestinal enzymes play an important part in this process. The physiology and biochemistry of the digestion system of termites from the genus Anacanthotermes is still not fully studied. In the present research, we studied the activity of some carbohydrases in termites’ salivary glands. Our data show that the activity of exocellulase in adult termites is 1.5 times more that in young individuals and 3 times more active than in nymphs, while the exocellulase in soldiers remains inactive. Moreover, the activity of celluloses in the intestine of A. turkestanicus is still not fully studied. We observed that exocellulase is involved in the digestion of food polymers in the castes of termites-workers, nymphs and soldiers. Thus, cellulase activity in the intestinal tissue (intrinsic) is manifested at a very low level in all termites’ castes we studied. However, the activity of exocellulase in the intestinal cavity (symbiotic) increases 3.5 times in workers, 3.0 times in nymphs and 2.4 times in soldiers compared to that in intestinal tissue. In addition, the intrinsic activity of this enzyme differed little in all three castes of termites, whereas the symbiotic activity in workers and nymphs was more showed than in soldiers.

Periodontitis and proteases. Hardly a closed case

During periodontal inflammation, a multitude of substances both from the host and of microbiological origin are released into the gingival crevicular fluid (GCV) and saliva. These substances, such as proteins and peptides, can therefore be regarded as biomarkers of the inflammatory process. With the help of sensitive and advanced laboratory technologies, the role of these biomarkers has been identified. However, the high costs, complexity and difficulty of interpretation of the results often hinder the introduction of biomarkers for diagnostic purposes in the dental practice. Certain salivary enzymes, proteases, can function as biomarkers and haveinteresting properties for producing rapid chairside diagnoses, because the presence of a protease or proteolytic activity can be demonstrated in a simple and rapid biochemical way, for example by colour signalling. Since other processes in the oral cavity influence the results of the test, such tests are especially useful as one element in a broader oral diagnostic investigation.

Feline dental implants: new paradigm shift in maxillary alveolar osteitis treatment planning with osseodensification

Background Felines are obligate carnivores and use their teeth for prehending as well as tearing and dissecting their food. Mastication is the first step of digestion, aiding in the lubrication of the food and the formation of a bolus. Mastication also increases the surface area of the food to be initially digested by salivary enzymes before being more easily swallowed. The teeth are specialized structures which play an important role in mastication, grooming, supporting the lips and the tongue, as well as being used as weapons for hunting and for self-defense. Although some mammals can still survive with few or no teeth at all, cats included, losing teeth can influence the types of foods that can be eaten, and will affect the ability to masticate and processing of the food prior to swallowing. Dental implants have become an option for replacement of lost canines in these animals. Although cats appear to manage well after a canine extraction, complications such as lip entrapment can occur, especially after maxillary canine tooth extraction. Even cats with a complicated crown fracture of the maxillary canine tooth that have had root canal therapy to preserve the tooth, can still suffer from lip entrapment. This can lead to painful lip ulcers and the need for further dental treatment. Today, canine tooth replacement with a dental implant/crown is a predictable option that can be offered to clients who would like to replace a lost canine tooth. Case report Two long term cases and follow up of lost maxillary canines caused by alveolar osteitis and replacement by dental implants, to restore normal function in feline subjects are reported. Conclusion Dental implants proved to be a viable long-term treatment option in the 2 cats described.Immediate implant placement at the time of extraction helps to maintain the surrounding bone structures and prevent collapse of the labial buccal plate. So far, the authors have placed 30 implants in 15 feline subjects up to a 3-year follow-up and no integration failures have been observed to date.

Association between salivary amylase enzyme activity and obesity in Saudi Arabia.

Obesity is a significant public health concern that predisposes individuals to a high risk of premature mortality. Previous studies also reported that low serum concentrations of AMY1 have been associated with obesity. The aimed of the study to assess the relationship between salivary amylase (AMY1) activity and body mass index (BMI) in Saudi male and female adults in Riyadh. This study included a total of 200 (100 individuals who were overweight and obese and 100 who had normal body weight [control individuals]) Saudi participants aged 20 to 50 years old. They were recruited from physical fitness clubs and were school employees in Riyadh City. The dietary food intake was assessed using a 24-hour dietary recall. The activity of the AMY1 was measured using a microplate fluorescence reader. A significant (P ≤ .05) increase was observed in the incidence of hypertension, dyslipidemia, diabetes mellitus (DM), and family history of overweight and obesity in overweight and obese individuals than in the control individuals, and these were in parallel to the significant increase in weight, waist circumference (WC), hip circumference (HC), systolic blood pressure (SBP), diastolic blood pressure (DBP), and BMI. A significant (P ≤ .05) increase was also observed in the carbohydrate and total fat dietary intake of overweight and obese individuals in relation to the respective dietary reference intake (DRI) values. AMY1 activity was significantly lower than the reference values in the overweight and obese group. Furthermore, AMY1 activity was significantly (P ≤ .05) reverse with weight, WC, HC, and BMI in both males and females in the overweight and obese group. In conclusion, the Saudi overweight and obese population seems to be at risk of low AMY1, which correlates with their obesity.

Biomarker- A natural boon for the artificial burden

Biomarker- A natural boon for the artificial burden - IJOHD- Print ISSN No: - 2395-4914 Online ISSN No:- 2395-499X Article DOI No:- 10.18231/j.ijohd.2019.005, International Journal of Oral Health Dentistry-Int J Oral Health Dent

Biofilm composition and composite degradation during intra-oral wear

ObjectivesThe oral environment limits the longevity of composite-restorations due to degradation caused by chewing, salivary and biofilm-produced enzymes and acids. This study investigates degradation of two resin-composites in relation with biofilm composition in vitro and in vivo. MethodsSurface-chemical-composition of two Bis-GMA/TEGDMA composites was compared using X-ray-Photoelectron-Spectroscopy from which the number ester-linkages was derived. Composite-degradation was assessed through water contact angles, yielding surface-exposure of filler-particles. Degradation in vitro was achieved by composite immersion in a lipase solution. In order to evaluate in vivo degradation, composite samples were worn in palatal devices by 15 volunteers for 30-days periods in absence and presence of manually-brushing with water. PCR-DGGE analysis was applied to determine biofilm composition on the samples, while in addition to water contact angles, degradation of worn composites was assessed through surface-roughness and micro-hardness measurements. ResultsIn vitro degradation by lipase exposure was highest for the high ester-linkage composite and virtually absent for the low ester-linkage composite. Filler-particle surface-exposure, surface-roughness and micro-hardness of both resin-composites increased during intra-oral wear, but filler-particle surface-exposure was affected most. However, based on increased filler-particle surface-exposure, the high ester-linkage composite degraded most in volunteers harvesting composite biofilms comprising Streptococcus mutans, a known esterase and lactic acid producer. This occurred especially in absence of brushing. SignificanceDegradation during intra-oral wear of a low ester-linkage composite was smaller than of a high ester-linkage composite, amongst possible other differences between both composites. S. mutans herewith is not only a cariogenic, but also a composite-degradative member of the oral microbiome.

Determining damage levels in wheat caused by Sunn pest (Eurygaster integriceps) using visible and near-infrared spectroscopy

A commercial near-infrared (NIR) instrument for bulk samples and a modified Single Kernel Near-Infrared (SKNIR) instrument equipped with a visible or NIR spectrometer were studied as a way to measure damage levels caused by Sunn pest (SP) in wheat. Sunn pest causes damage by feeding on wheat berries and injecting a salivary enzyme damaging the gluten. For measurement of SP damage in bulk wheat, NIR calibration models developed for mixtures containing 0–10% and 0–100% sound and SP damaged wheat resulted in R2 of 0.25 and 0.89 and SECVs of 2.75 and 10.9, respectively. The 0–100% model was considered a qualitative measure of damage, but predictions were poor over 0–10% which is a critical range for commercial applications. Discrimination between single kernels of Sound and SP-damaged was typically good, with classification accuracy averaging ∼75% for both visible and NIR although some were poor, which greatly affected the average. Average classification accuracy was ∼85% for spectral data that contained kernels from all samples. While the potential for using visible or NIR spectroscopy was shown, results highlighted the need to develop a more robust SP classification model to further evaluate the single kernel model.

Efficacy of salivary and diastase extracts of Piper betle in modulating the cellular stress in placental trophoblast during preeclampsia

Background: Betle leaf (BL) is an ancient herb with a potential nontoxic natural antioxidant efficacy. Oral intake and chewing betel leaves have an effort on the moving parts of salivary gland and induce salivation and detoxification; hence plays a vital role in oral hygiene. BL produces and activates various salivary enzymes which include α-amylase, lipase, lysozyme, and lactoperoxidase. BL possess antibacterial, antioxidant, anti-diabetic, immunomodulatory, and antihypertensive properties. Hence, it can be effective in the treatment of hypertensive disorder of pregnancy like preeclampsia (PE). Objective: In this context, two extracts of BL (Salivary BL extract [SBLE] and diastase BL extract [DBLE]), proved to impart efficient radical scavenging activity in our previous research work were utilized to analyze their efficacy on the level of 4-hydroxy nonenal (4-HNE), heme oxygenase-1 (HO-1), asymmetric dimethylarginine (ADMA), adiponectin/leptin in the trophoblast isolated from placental tissue of both normotensive and preeclamptic patients. Materials and Methods: Trophoblasts were isolated from both subjects and incubated with different extracts of Piper betle (saliva and diastase) to assess the cellular stress in placental trophoblast during PE. Results: Results demonstrate that extracts of BL exhibit a significant role in regulating 4-HNE, ADMA, leptin, HO-1, and adiponectin in preeclamptic trophoblasts. Still, SBLE showed relatively higher efficiency in defining the level of 4-HNE, ADMA, leptin, HO-1, and adiponectin in preeclamptic trophoblasts than DBLE, suggesting that it may be attributed to the interaction of the phytochemicals present in BL with the components of saliva. Conclusion: Hence, BL may be recommended as the effective natural remedy in the management of pathological complications of PE; which may preclude low birthweight babies and ensure live fetal delivery. Abbreviations Used: 4-HNE: 4-hydroxy-nonenal, ADMA: Asymmetric dimethylarginine, BL: Betel leaf, BLE: Betel leaf extract, CO: Carbon monoxide, DB: Diastase with betel leaf, DBLE: Diastase betel leaf extract, ELISA: Enzyme-linked immunosorbent assay, eNOS: Endothelial nitric oxide synthase, FBS: Fetal bovine serum, GSH: Reduced glutathione, HBSS: Hank's balanced salt solution, HO-1: Heme oxygenase-1, iNOS: Inducible nitric oxide synthase, NO: Nitric oxide, NOS: Nitric oxide synthase, PE: Preeclampsia, ROS: Reactive oxygen species, SB: Saliva with betel leaf, SBLE: Salivary betel leaf extract.

Salivary Cortisol Level; Its Role in Stress Induced in Patients Undergoing Dental Treatment; An in Vivo Study

Aims and objective: The stress control is essential in patients to elude the loss of motivation for Dental procedures. The present study was conducted to estimate salivary cortisol response to stress developed in patients undergoing Dental treatment Methodology: The salivary cortisol level were evaluated in 40 patients ,out of which 10 patients were healthy patient who do not require any dental treatment and 40 patients were the study group who underwent scaling,implant treatment and class 1 cavity preparation for restoration .The groups demarcated were as follows. Group 1: control group with no treatment and just the saliva sample was taken. Group 2: The patients undergone scaling Group 3: The patients undergone implant treatment Group 4: The patients undergone class 1 cavity preparation for restoration. The salivary cortisol levels were evaluated using Salivary Cortisol Enzyme Immunoassay Kit. Results: The salivary cortisol level was highest in Group 3,followed by Group 2 ,Group 4 and least in Group 1. Conclusion: The stress level was lowest in healthy patient and highest when patient underwent implant placement in appointment.

Salivary antioxidant enzymes and lipid peroxidation product malondialdehyde and sialic acid levels among smokers and non-smokers with chronic periodontitis-A clinico-biochemical study.

Background:Pathogenesis of most of the inflammatory process are associated with reactive oxygen species (ROS), derived from various metabolic sources and which may lead to direct or indirect tissue damage due to oxidative stress, resulting in periodontal diseases. Usually antioxidant systems are capable of removing free radicals, thereby preventing tissue damage from free radical. ROS can result in tissue damage, involving lipid peroxidation. The aim of this study was to evaluate and compare the levels of superoxide dismutase (SOD), glutathione peroxidase (GPx), malondialdehyde (MDA), and sialic acid (SA) in periodontally healthy and chronic periodontitis among nonsmokers and smokers and to determine their value as diagnostic markers for chronic periodontitis.Materials and Methods:A total of 90 male patients aged 20--60 years were recruited and grouped as Group 1: 30 Healthy nonsmokers, who had never smoked. Group 2: 30 nonsmokers with chronic periodontitis. Group 3: 30 smokers with chronic periodontitis. Unstimulated saliva was collected for at least 5 min and clinical measurements; SOD, GPx, MDA and SA were assessed using a spectrophotometric method.Results:Data showed a significant correlation between salivary SOD, GPx, MDA, and SA in group 1, group 2, and group 3. SOD and GPx were found to be lower and MDA and SA levels were found to be higher among smokers with chronic periodontitis.Conclusion:Reduced levels of antioxidant enzymes SOD and GPx and elevated levels of lipid peroxidation product MDA as well as increased levels of SA could be used as diagnostic markers to measure oxidative stress in periodontal disease associated with risk factor such as smoking.

Effects of beta-cypermethrin and myclobutanil on some enzymes and changes of biomarkers between internal tissues and saliva in reptiles (Eremias argus)

Numerous studies suggested that reptiles are sensitive to environmental pollution and the abundance of many species are in decline. Our research is aim to assess the toxic effects of pesticide in reptiles. And we also want to supply some data about nondestructive samples for environmental risk assessment in reptiles. Lizards were orally administered a single-dose of beta-cypermethrin (BCP) or myclobutanil (MC) at the concentration of 20 mg/kg body weight (bw). The results showed that pesticides could induce changes in enzymatic activities (SOD, CAT, LDH, AChE) and MDA levels in organs or tissues of lizards. BCP could cause more severe oxidative damage than that of MC. Salivary enzymes activities showed sensitivity changes to the toxicity of pesticides. We could use saliva to reflect whether the reptiles are toxic by pesticides. We also agree that buccal swabs could be used as a tool for saliva sampling.

Evaluation of salivary cortisol levels in relation to dento-maxillary prosthesis adjustment

PurposeThe purpose of this study was to investigate the influence of dento-maxillary prosthesis adjustment procedure on levels of salivary cortisol. MethodsNine participants (six men, three women, mean age 65.9 years) took part in this study. Saliva samples were collected before and after dento-maxillary prosthesis adjustment during the four different visits. Free cortisol levels were determined using a salivary cortisol immunoassay kit (expanded-range high-sensitivity salivary cortisol enzyme immunoassay kit, Salimetrics). Besides, original self-report sheets, a 35-item food intake questionnaire, the University of Washington Quality of Life (UW-QOL) questionnaire version 4, and the Geriatric Oral Health Assessment Index (GOHAI) questionnaire were also administered. The changes of salivary cortisol levels were analyzed using 2-level multilevel linear regression, with adjustment for age, sex, and time. Wilcoxon signed-rank test was used to compare scores of the food intake questionnaire, UW-QOL questionnaire, and GOHAI questionnaire. ResultsSalivary cortisol levels decreased significantly after carrying out the dento-maxillary prosthesis adjustment procedure. During the third adjustment, the salivary cortisol levels were significantly low. In addition, salivary cortisol levels of participants aged 70 years and over were significantly higher than other aged groups. The total scores for grade III–V of the food intake questionnaire increased significantly. Other questionnaires had a trend toward increasing scores, yet the differences were not significant. ConclusionsWithin the limitations of this study, the results suggest that a reduction in symptoms of discomfort may have an influence on the decrease of salivary cortisol levels in dento-maxillary prosthesis wearers.

Salivary proteins and peptides in the aetiology of caries in children: Systematic literature review.

To assess the relationship of chosen salivary proteins and peptides levels with the occurrence of caries in children. PubMed, MEDLINE and EMBASE databases from 2000 to 2018 were researched for original observational studies published in English. The risk of bias and quality of the included papers were assessed regarding the guidelines by Fowkes and Fulton. Twenty-two studies were included in the review, from which the issue of glycoproteins (including immunoglobulins), AMPs and salivary enzymes was discussed. The research involved primary dentition (13 papers), as well as mixed (7) and permanent dentition (5). Caries assessment included visual inspection, dmft/s and DMFT/S indexed; quantity of Streptococcus mutans and Lactobacillus spp. bacteria; and caries risk assessment. The results of studies regarding the connection between salivary peptides and proteins and caries development in children are promising; however, further investigations should be undertaken. The majority of studies included are case-control and cross-sectional; however, it is necessary to conduct more cohort studies with adequate follow-up prior to considering this as markers for caries risk assessment.

Salivary lactate dehydrogenase (LDH) as a tool for early diagnosis of oral cancer in individuals with Fanconi anemia

Currently one of the greater challenges is the diagnosis and treatment of cancer. Many studies address the genetic and metabolic aspects to support in early diagnosis and increase the survival of individuals at high risk. Individuals with Fanconi anemia can be included in this high risk group because they have a predisposition to develop head and neck cancer. The use of salivary enzymes as biomarkers to detect the changes in oral tissue at the initial phase seems viable, because saliva is easy to obtain, it moisture oral mucosa and cells metabolic compounds can be found on it. Due to the metabolic characteristics of the cancer cell, an increase in Lactate Dehydrogenase (LDH) may indicate a carcinogenesis process. The hypothesis of this study is to use of salivary LDH as a tool in the early diagnosis of oral cancer on a high risk group such as Fanconi anemia’s patients.

Emulsion Microgel Particles as High-Performance Bio-Lubricants.

Starch-based emulsion microgel particles with different starch (15 and 20 wt %) and oil contents (0–15 wt %) were synthesized, and their lubrication performance under physiological conditions was investigated. Emulsion microgels were subjected to skin mimicking or oral cavity mimicking conditions, i.e., smooth hydrophobic polydimethylsiloxane ball-on-disc tribological tests, in the absence or presence of salivary enzyme (α-amylase). In the absence of enzyme, emulsion microgel particles (30–60 vol % particle content) conserved the lubricating properties of emulsion droplets, providing considerably lower friction coefficients (μ ≤ 0.1) in the mixed lubrication regime compared to plain microgel particles (0 wt % oil). Upon addition of enzyme, the lubrication performance of emulsion microgel particles became strongly dependent on the particles’ oil content. Microgel particles encapsulating 5–10 wt % oil showed a double plateau mixed lubrication regime having a lowest friction coefficient μ ∼ 0.03 and highest μ ∼ 0.1, the latter higher than with plain microgel particles. An oil content of 15 wt % was necessary for the microgel particles to lubricate similarly to the emulsion droplets, where both systems showed a normal mixed lubrication regime with μ ≤ 0.03. The observed trends in tribology, theoretical considerations, and the combined results of rheology, light scattering, and confocal fluorescence microscopy suggested that the mechanism behind the low friction coefficients was a synergistic enzyme- and shear-triggered release of the emulsion droplets, improving lubrication. The present work thus demonstrates experimentally and theoretically a novel biolubricant additive with stimuli-responsive properties capable of providing efficient boundary lubrication between soft polymeric surfaces. At the same time, the additive should provide an effective delivery vehicle for oil soluble ingredients in aqueous media. These findings demonstrate that emulsion microgel particles can be developed into multifunctional biolubricant additives for future use in numerous soft matter applications where both lubrication and controlled release of bioactives are essential.

Rapid Stain Biomarker : A Novel Tool for the Forensic Detection of Saliva (Effect of Time and Surface on its Reliability )

Saliva is one of the vital fluids secreted by the human body; it can be deposited on the human skin or other materials through biting, sucking, licking and kissing. Saliva stains encountered in forensic casework is an important source of DNA; so, it is considered an important evidence for personnel identification. The rapid Stain Identification of Human Saliva (RSIDTM-Saliva) bioassay is designed to detect specifically the presence of human salivary µ-amylase enzyme. Test development is completed within 10 minutes and can detect as little as 1ml of human saliva. The detection protocol can be completely integrated into the procedures for DNA extraction and analysis. Aim: Assess the efficacy of RSIDTM-Saliva strip test for the detection of human saliva under some different variables (different surfaces & different time intervals).Evaluate the quality of DNA extracted from unpreserved saliva and the possibility of its usage as evidence in forensics. Methodology:Fifty two saliva samples were collected from four volunteers. Forty eight of them were deposited over surfaces of different nature (plastic bottles, glass cup and cigarette butts) at the same time. Four saliva samples were collected directly from the floor of the mouth of each participant by a cotton swab (positive control group). All samples were left to dry for 10 min at room temperature then extracted and analyzed according to the protocol designed for (RSIDTM-Saliva). Collected samples were studied as three tested groups and as control groups. Saliva samples collected from each participant were categorized into four subgroups according to the time interval of saliva extraction [zero (on the spot), 10, 20, and 30 days].Positive samples for saliva identification were subjected to DNA extraction and typing; as three autosomal primers short tandem repeats (STRs) specific to human genomic DNA (D21S11, D18S51 and CSF1PO) were amplified by polymerase chain reaction (PCR); then subjected to separation and analysis of fragments size by agarose gel electrophoresis to determine their allelic size through comparison to the standard allelic 100 bp ladder. Results: Saliva could be identified in 100% of tested samples as well as for the positive control samples, which were subjected to successful DNA extraction followed by PCR amplification of the chosen three STRs. Results of agarose gel revealed that the sizes of these PCR products were in accordance with the 100 bp DNA ladder; validating the good quality and quantity of the extracted DNA Conclusion: the current study demonstrated that non-stored saliva deposited over different surfaces for up to 30 days presents an ideal source for DNA which may be used for forensic identification. The study also evidenced that the new RSID-saliva strip test is a fast, easy, sensitive and reliable method for saliva identification over different surfaces.

Immunity to LuloHya and Lundep, the salivary spreading factors from Lutzomyia longipalpis, protects against Leishmania major infection.

Salivary components from disease vectors help arthropods to acquire blood and have been shown to enhance pathogen transmission in different model systems. Here we show that two salivary enzymes from Lutzomyia longipalpis have a synergist effect that facilitates a more efficient blood meal intake and diffusion of other sialome components. We have previously shown that Lundep, a highly active endonuclease, enhances parasite infection and prevent blood clotting by inhibiting the intrinsic pathway of coagulation. To investigate the physiological role of a salivary hyaluronidase in blood feeding we cloned and expressed a recombinant hyaluronidase from Lu. longipalpis. Recombinant hyaluronidase (LuloHya) was expressed in mammalian cells and biochemically characterized in vitro. Our study showed that expression of neutrophil CXC chemokines and colony stimulating factors were upregulated in HMVEC cells after incubation with LuloHya and Lundep. These results were confirmed by the acute hemorrhage, edema and inflammation in a dermal necrosis (dermonecrotic) assay involving a massive infiltration of leukocytes, especially neutrophils, in mice co-injected with hemorrhagic factor and these two salivary proteins. Moreover, flow cytometry results showed that LuloHya and Lundep promote neutrophil recruitment to the bite site that may serve as a vehicle for establishment of Leishmania infection. A vaccination experiment demonstrated that LuloHya and Lundep confer protective immunity against cutaneous leishmaniasis using the Lu. longipalpis—Leishmania major combination as a model. Animals (C57BL/6) immunized with LuloHya or Lundep showed minimal skin damage while lesions in control animals remained ulcerated. This protective immunity was abrogated when B-cell-deficient mice were used indicating that antibodies against both proteins play a significant role for disease protection. Rabbit-raised anti-LuloHya antibodies completely abrogated hyaluronidase activity in vitro. Moreover, in vivo experiments demonstrated that blocking LuloHya with specific antibodies interferes with sand fly blood feeding. This work highlights the relevance of vector salivary components in blood feeding and parasite transmission and further suggests the inclusion of these salivary proteins as components for an anti-Leishmania vaccine.

Relation between salivary cortisol and alpha amylase levels and anxiety in Egyptian patients with minor recurrent aphthous stomatitis

Background/Aim: Recurrent aphthous stomatitis (RAS) is the most prevalent ulcerative disorder of the oral mucous membrane. The aim of the present contemplate is to assess salivary cortisol (SC) and salivary alpha amylase enzyme (SAA) levels in relation to anxiety in Egyptian patients with minor recurrent aphthous stomatitis. Subjects and Methods: patients with minor RAS as a study group (RAS group, n = 35) and healthy subjects as a control (control group, n = 35) who were matching regarding age, sex, and socioeconomic state were included in this study. SC and SAA were assessed in all participants using enzyme-linked immunosorbent assay (ELISA) and kinetic enzyme assay respectively. Hamilton’s anxiety scale (HAS) has been used for evaluation of anxiety level in both study groups.Results: A significantly high mean SC (1.7 ± 0.34 μg/dl), SAA (137.87 ± 47.16 U/ml) levels and anxiety score (26.7 ± 2.21) were recorded in RAS group (p = 0.00, 0.02, 0.00) respectively in comparison to control group.Conclusion: Anxiety may have a pivotal role in etiopathogensis of RAS hence; Psychological management should be considered in RAS treatment.