Oral cavity protozoa relevant in the practice of dentistry

Introduction: Oral cavity is the colonization site of most diverse microorganisms. The homeostasis of oral microbioma is affected by numerous factors. Periodontal diseases occur as a consequence of disturbed oral microbioma homeostasis, when an inflammatory reaction occurs in the periodontal tissue. The impact of parasites on periodontal pathophysiology has not been sufficiently studied, and present and future research should hopefully answer quite a few questions concerning the issue. Aim of the paper: In the light of the present knowledge of the pathogenesis, diagnosis and epidemiology of oral cavity infections caused by Entamoeba gingivalis and Trichomonas tenax, the aim of the paper was review of literature which could to point to the importance of protozoa in the practice of dentistry and to possible oral cavity manifestations of parasitic infections relevant for public health. Conclusion: Dentists have an essential role in the diagnosis of oral diseases caused by oral cavity protozoa, and protozoa relevant for public health that produce systemic infections, the pathological changes of which may manifest in the oral cavity. Their identification represents a challenge and requires multidisciplinary approach for a timely diagnosis and adequate management

Soil and sand contamination with canine intestinal parasite eggs as a risk factor for human health in public parks in Niš (Serbia)

Regarding geographical distribution and clinical relevance, the most common canine geohelminths are Toxocara canis, ancylostomatids, and Trichuris vulpis. Canine intestinal parasites from the soil and sand present an important potential serious human health hazard, especially for the children preschool and school – age. This paper aimed to establish the degree of contamination of soil and sand with zoonotic parasites from the canine feces and the degree of risk they could pose for human health in public places and playgrounds in the city of Niš. Our parasitological study involved 200 soil samples and 50 sand samples from the public parks in the city of Niš in southeastern Serbia (43°19′15″N, 21°53′45″ E). From several locations, about 100 g of soil and sand was collected based on the bioclimatic indices. Parasitological diagnosis was performed using conventional qualitative and quantitative coprological methods, abiding by the recommendations about the diagnosis of parasitic diseases. In 38 – 46 % of soil samples and 40 % of sand samples seven species of endoparasites were diagnosed. In the samples of soil, a medium and high degree of contamination with the ascarid T. canis (14 – 22 %) was detected, as well as a low and medium degree of contamination with ancylostomatids (4 – 12 %), and in the samples of sand, a variable degree of contamination with the helminths T. canis (26 %) and A. alata (16 %) was found. A statistically significant difference was found in the contamination with A. alata eggs between the samples of sand and samples of soil. The studied public surfaces represent the reservoir of zoonotic parasites, which is a public health problem requiring a synergistic action of several factors to be successfully resolved, i.e. the implementation of prevention, surveillance, and control measures

Poster presentation: 1H-NMR-based serum metabolomics of bipolar disorder patients

Bipolar disorder (BD) is a mental disorder that causes alteration of mood states including mania, depression, and euthymia and it is ranked as one of the leading causes of disability and premature mortality, with a prevalence of 60 million people worldwide. BD is a heterogenous illness including diverse genetic, environmental, and biochemical factors and its pathophysiology is still largely unknown. Diagnosis of BD exclusively depends on the subjective recognition of symptoms without any objective methods such as a clinical test of biomarker identification, instigating misdiagnosis, inadequate treatments and deficient clinical outcomes. 1H-NMR-based serum metabolomics of Serbian patients with BD (33) and healthy controls (39) contributed to identification of 22 metabolites for this disease. Threonine, aspartate, gamma-aminobutyric acid, 2-hydroxybutyric acid, serine, and mannose make a unique biomarker set, and were confirmed for the first time in BD Serbian serum samples. Additional six identified metabolites (3-hydroxybutyric acid, arginine, lysine, tyrosine, phenylalanine, and glycerol) are in accordance with the previously determined NMR-based sets of serum BD biomarkers in Brazilian and/or Chinese patient samples, while nine identified metabolites (lactate, alanine, valine, leucine, isoleucine, glutamine, glutamate, glucose, and choline) are the same established biomarkers in three different ethnic and geographic origins (Serbia, Brazil, and China). The same confirmed metabolites are an indicator of the right path in discovery of the universal set of BD biomarkers by NMR.Poster presented at 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK, September 4-6, 2023, Belgrade, SerbiaAbstract:[https://cer.ihtm.bg.ac.rs/handle/123456789/7390

1H-NMR-based serum metabolomics of bipolar disorder patients

Bipolar disorder (BD) is a mental disorder that causes alteration of mood states including mania, depression, and euthymia and it is ranked as one of the leading causes of disability and premature mortality, with a prevalence of 60 million people worldwide. BD is a heterogenous illness including diverse genetic, environmental, and biochemical factors and its pathophysiology is still largely unknown. Diagnosis of BD exclusively depends on the subjective recognition of symptoms without any objective methods such as a clinical test of biomarker identification, instigating misdiagnosis, inadequate treatments and deficient clinical outcomes. 1H-NMR-based serum metabolomics of Serbian patients with BD (33) and healthy controls (39) contributed to identification of 22 metabolites for this disease. Threonine, aspartate, gamma-aminobutyric acid, 2-hydroxybutyric acid, serine, and mannose make a unique biomarker set, and were confirmed for the first time in BD Serbian serum samples. Additional six identified metabolites (3-hydroxybutyric acid, arginine, lysine, tyrosine, phenylalanine, and glycerol) are in accordance with the previously determined NMR-based sets of serum BD biomarkers in Brazilian and/or Chinese patient samples, while nine identified metabolites (lactate, alanine, valine, leucine, isoleucine, glutamine, glutamate, glucose, and choline) are the same established biomarkers in three different ethnic and geographic origins (Serbia, Brazil, and China). The same confirmed metabolites are an indicator of the right path in discovery of the universal set of BD biomarkers by NMR

Poster presentation: 1H-NMR metabonomic view on schizophrenia

Schizophrenia (SCZ) is a brain disease leading to significant functional impairments and premature death, and it affects 20 million people worldwide. Due to the complexity of this disease including different genetic and environmental factors, there is a lack in understanding pathophysiology and diagnosis of schizophrenia. In order to overcome existing gaps, the establishment of a universal set of SCZ biomarkers has a crucial role. Metabonomic study of serum samples of Serbian patients with schizophrenia (51) and healthy controls (39) by 1H-NMR analyses associated with chemometrics, provided the identification of 26 metabolites/biomarkers for this disorder. The biomarker set including aspartate/aspartic acid, lysine, 2-hydroxybutyric acid, and acylglycerols was established for the first-time in SCZ serum samples of Serbian patients by 1H-NMR experiments. The other 22 identified metabolites are in agreement with the previously confirmed NMR-based serum biomarker sets of Brazilian and/or Chinese patient samples. The same 13 metabolites (lactate/lactic acid, threonine, leucine, isoleucine, valine, glutamine, asparagine, alanine, gamma-aminobutyric acid, choline, glucose, glycine and tyrosine) were established in all SCZ samples from three countries of different ethnicity and geographical origins (Serbia, Brazil and China). These results emphasize the crucial role in the possibility of their application as biomarkers for diagnosis of SCZ, reliable monitoring of treatment response and clinical outcomes.Poster presented at 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK, September 4-6, 2023, Belgrade, SerbiaAbstract: [https://cer.ihtm.bg.ac.rs/handle/123456789/7392

1H-NMR metabonomic view on schizophrenia

Schizophrenia (SCZ) is a brain disease leading to significant functional impairments and premature death, and it affects 20 million people worldwide. Due to the complexity of this disease including different genetic and environmental factors, there is a lack in understanding pathophysiology and diagnosis of schizophrenia. In order to overcome existing gaps, the establishment of a universal set of SCZ biomarkers has a crucial role. Metabonomic study of serum samples of Serbian patients with schizophrenia (51) and healthy controls (39) by 1H-NMR analyses associated with chemometrics, provided the identification of 26 metabolites/biomarkers for this disorder. The biomarker set including aspartate/aspartic acid, lysine, 2-hydroxybutyric acid, and acylglycerols was established for the first-time in SCZ serum samples of Serbian patients by 1H-NMR experiments. The other 22 identified metabolites are in agreement with the previously confirmed NMR-based serum biomarker sets of Brazilian and/or Chinese patient samples. The same 13 metabolites (lactate/lactic acid, threonine, leucine, isoleucine, valine, glutamine, asparagine, alanine, gamma-aminobutyric acid, choline, glucose, glycine and tyrosine) were established in all SCZ samples from three countries of different ethnicity and geographical origins (Serbia, Brazil and China). These results emphasize the crucial role in the possibility of their application as biomarkers for diagnosis of SCZ, reliable monitoring of treatment response and clinical outcomes

NMR Metabolomics in Serum Fingerprinting of Schizophrenia Patients in a Serbian Cohort

Schizophrenia is a widespread mental disorder that leads to significant functional impairments and premature death. The state of the art indicates gaps in the understanding and diagnosis of this disease, but also the need for personalized and precise approaches to patients through customized medical treatment and reliable monitoring of treatment response. In order to fulfill existing gaps, the establishment of a universal set of disorder biomarkers is a necessary step. Metabolomic investigations of serum samples of Serbian patients with schizophrenia (51) and healthy controls (39), based on NMR analyses associated with chemometrics, led to the identification of 26 metabolites/biomarkers for this disorder. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) models with prediction accuracies of 0.9718 and higher were accomplished during chemometric analysis. The established biomarker set includes aspartate/aspartic acid, lysine, 2-hydroxybutyric acid, and acylglycerols, which are identified for the first time in schizophrenia serum samples by NMR experiments. The other 22 identified metabolites in the Serbian samples are in accordance with the previously established NMR-based serum biomarker sets of Brazilian and/or Chinese patient samples. Thirteen metabolites (lactate/lactic acid, threonine, leucine, isoleucine, valine, glutamine, asparagine, alanine, gamma-aminobutyric acid, choline, glucose, glycine and tyrosine) that are common for three different ethnic and geographic origins (Serbia, Brazil and China) could be a good start point for the setup of a universal NMR serum biomarker set for schizophrenia

Evaluation of the universality of NMR metabolic fingerprints of schizophrenia

Schizophrenia (SCZ) is a very disabling mental disorder whose molecular basis is a combination of many factors still not completely understood, with a diagnosis based on observed behavior, the person's reported experiences and reports of others that are familiar with the person, with no objective test. Also, up to date, there are no reliable markers for monitoring the SCZ. NMR-metabolomics [1] reported in 2017 bring some of the possible markers from blood serum of SCZ individuals linked strongly with known dopamine, glutamate and GABA dysfunction in SCZ. As to verify if these findings are universal, we have compared the SCZ patients from geographically different environments and cited interesting SCZ characteristics. The first set of samples was collected in Belgrade, Serbia. 14 mental health patients (50% male) with 52.86 ± 7.27 years of age had a confirmed diagnosis of SCZ. The control group of 13 healthy individuals (69% male) had none of psychotic disorders, and individuals were 23.07 ± 2.79 years of age. Blood serum samples were collected and prepared for the analysis following the published methodology [1, 2]. NMR spectra were measured on a Bruker AVANCE III spectrometer (500.26 MHz for 1H). The spectra were acquired at 298 K with 128 scans and 32 k. The serum samples were prepared and measured as triplicates. On the other side, the group of individuals from Brazil that was matched in number, age, gender and history of mental illness with individuals from Serbia was previously described [1]. 1H NMR spectra were phase and baseline corrected using MestreNova and the lactate doublet was used as the chemical shift reference. The data were binned (0.005 ppm) in a spectral range 0.50 - 9.00 ppm, while the residual HDO peak (4.50-5.00 ppm) was excluded. Then, the data were normalized by the sum equal to 1000, the variables were mean centered and PCA and PLS-DA were performed using MATLAB. It was shown that the mental health patients have clearly different blood serum metabolites when compared to the healthy ones independently from where the samples were obtained with almost identical marker set. Also, it was shown that the samples are different metabolically when Brazilian and Serbian samples were compared. 1] L. Tasic et al., Schizophrenia Research 2017, 185, 182. [2] J. Pontes et al., Analytical Methods 2017, 9, 1078

DIROFILARIA REPENS – CAUSE OF ZOONOSIS

Dirofilariasis is zoonosis caused by genus Dirofilaria. The common couse of this zoonosis is species Dirofolaria repens (D. repens). This species is natural parasites of dogs, but in the last years in our country, sporadic cases of human dirofilariasis coused by D. repens have been described. Pathological and clinical sings in infected dogs are non-characteristic. This species lives in subcutaneous tissue, the space between ligaments and blood vessels, in kidney and lymphatic nodes. In humans, the most common sites are conjuctiva and subcutaneous tissue. Diagnosis of infection in dogs is based on anamnestic data, clinical signs and results of specific diagnostic tests for detection of microfilariae in peripheral blood. The therapy is aimed at destruction of adult forms and microfilariae in the peripheral blood of dogs. Mechanical or chemical methods and drugs can be used as prophylactic measures against filariasis in dogs. With geographical spreading of D. repens infection in dogs, we can inspect more cases of human dirofilariasis. Therefore, we should put an accent on continuous control, follow-up of prevalence and distribution of this infection in dogs, which must be considered as a health problem

Seroprevalence of dirofilarioses in Serbia

In 2009 canine and human dirofilarioses were investigated in Serbia by means of home-made immunoassays to detect antibodies to D. repens and D. immitis somatic/metabolic polyproteins and, in dogs, to the recombinant Wolbachia surface protein (rWSP) (Marcos-Axutegi et al., 2005, Proc EMOP IX, 2: 297-302). A total of 122 dogs (56 males and 66 females; mean age = 4.15 years) were checked, coming from two northern areas of Serbia (Pančevo and Veliko Gradište). Considering their shelter, nutrition, care, training, working, health protection, sanitary controls, 38 of them were dogs bred in partially controlled life conditions and the remaining 84 subjects lived in uncontrolled life conditions. Human sera were taken from 298 people (109 males, 189 females) living in different areas of Serbia (Pančevo, Novi Sad, Zaječar, Leskovac, Vranje, Niš, Pirot). Antibodies to D. immitis were detected in 28 dogs, indicating an overall heartworm disease seroprevalence of 22.9%, whereas IgG antibodies to D.repens and to rWSP were evidenced in 42.6% and 5.7% of the subjects, respectively. As for the two different areas, antibodies to Dirofilaria were detected in 72.9% of dogs living in Pančevo, therefore more than living in Veliko Gradište (57.1%). No risk factors were evidenced, confirming data on the uselessness of prophylactic drugs at least against D. repens, and suggesting the presence, in these areas, of by day, sunrise or sunset biting mosquitoes as important vectors. As previously evidenced, the low antibody production to rWSP depends on the parasitic burden and on the larval/adult stage of the worms and to the cytokine synthesis patterns activated by this endosymbiont which is different from that activated by D. repens and D. immitis (Simon et al., 2009, Trends in Parasitology, 25 (9): 404-409). As for humans, 46 people (14 men and 32 women) had antibodies to dirofilarial nematodes, accounting for an overall seroprevalence of 15.4%. Particularly, antibodies to D. repens were found in 29 (9.7%) subjects, and that to D. immitis in 24 (8.1%) individuals; seven of them had specific antibodies to both species. No differences were evidenced by people sex (p=0.347), by dirofilarial species (p=0.551) and by area (p=0.056). However the highest seroprevalence were found in Pančevo (27,1%), one of area where we proved very high seroprevalence (72.9%) in dogs. In Serbia diriofilarioses represent an emerging disease of interest for both human and veterinary health. Infections are mainly due to D. repens, even if an active transmission level of both species has been evidenced. Our findings indicates that also D. immitis is transmitted to humans, although all the cases by now reported in the country have been attributed to D. repens. More widespread and detailed investigations are needed to better define the impact of this neglected disease on humans. Finally, entomological investigations are needed to identify the mosquito species actually involved in the parasites transmission and the vector efficiency of each of them, which are data essential to start appropriate control programs