Importance of NK Cells in Cellular and Humoral Responses Triggered by Pneumococcus Vaccination.

INTRODUCTION: Despite the success of vaccination in reducing overall rate of pneumococcal pneumonia, Streptococcus pneumoniae is still held responsible for high mortality and modality rates worldwide. Our study aimed to investigate the potential role played by NK cells in immune response generated by pneumococcal vaccination, which could contribute to the development of more effective vaccines. METHODS: The study included mice with and without NK cell depletion which were immunized with pneumococcus polysaccharide-conjugated vaccine followed by pneumococcus polysaccharide vaccine (PPV). Serum samples and splenocytes were collected from mice sacrificed 4 weeks after the last PPV dose. Serum samples were used for antibody level quantification by ELISA assay, while splenocytes were treated with PPV in vitro before monitoring CD4+ T-cell subsets (TH1, TH2, and TH17) and cytokine (IFN-γ, IL-4, and IL-17) secretion levels by flow cytometry and ELISA analysis, respectively. RESULTS: Results demonstrated reduced pneumococcal IgG and TH1 cell levels due to NK cell depletion. Nevertheless, in contrast to these observations, IFN-γ secretion levels after in vitro PPV-23 treatment of splenocytes did not exhibit any statistically significant difference between the two mice groups. CONCLUSIONS: The data indicate a positive contribution of NK cells to both T-cell and B-cell responses triggered against pneumococcal vaccination. Further studies are required to confirm our data and investigate the potential benefit of NK cell targeting in promoting vaccine efficacy, especially in the elderly population who continues to be affected significantly by pneumococcal pneumonia.

Prevalence of Blastocystis and Dientamoeba fragilis in diarrheal patients in Corum, Türkiye.

To investigate the prevalence of Blastocystis and Dientamoeba fragilis in diarrhea patients and healthy individuals in Corum, Türkiye, fecal samples from 92 diarrhea patients and 50 healthy individuals were collected and evaluated using direct microscopy and molecular methods to screen for bacteria, protozoa, and viruses. The prevalence of Blastocystis was 24.6% in total and more frequent in the healthy group (30.0%). The commonly detected STs (subtypes) were ST3 (40.0%) and ST2 (34.2%). The distribution of Blastocystis STs in the healthy and diarrheal groups did not show any difference in sex and age, but ST3 was detected more frequently in patients aged from 40 to 59 years (p < 0.05). Alleles 4 (8/12) and 2 (4/12) were present in ST1; 9 (3/5) and 12 (2/5) in ST2; 34 (9/14), 36 (3/14), and 38 (2/14) in ST3; and only allele 42 (2/2) in ST4. D. fragilis was present in 8.4% of the population. However, there was no statistically significant difference between the healthy and diarrheic groups (12.0% and 6.5%, respectively), neither with respect to age nor sex. Co-infection was 58.3% and was more frequent in healthy individuals (33.3%) than in diarrhea patients (25.0%). Blastocystis ST3 was the most common subtype detected, with D. fragilis at 33.3%. Salmonella, Shigella, or helminth eggs were not observed in all groups, but Entamoeba histolytica, Giardia intestinalis, Cryptosporidium, Rotavirus, Adenovirus, and Clostridium difficile toxin were found only in diarrhea patients. These findings support the hypothesis that Blastocystis and D. fragilis may be part of the healthy human gut microbiome.

[Outbreaks Due to Parasites: Examples from the World and Türkiye]. / Parazitlerden Kaynaklanan Salginlar: Dünyadan ve Türkiye'den Örnekler.

Outbreaks due to parasites can occur in various parts of the world and in different periods. These outbreaks can be caused by water and food, as well as by human-to-human or vector-borne transmission. Cryptosporidium spp. and Giardia intestinalis were among the pathogens that affected most people in water-borne outbreaks occurred in the world between 2010-2014. The chlorine resistance of both Cryptosporidium spp. and Giardia spp. leads to the widespread detection of these parasites in waterborne outbreaks. These two protozoans cause self-limiting watery diarrhea in immunocompetent individuals, but they can also cause chronic disease in certain situations. Apart from this, parasites such as Cyclospora spp., Cryptosporidium spp., Giardia intestinalis, Trichinella spp. and Toxoplasma gondii can also cause foodborne outbreaks. In Türkiye, outbreaks related to these parasites have emerged with the neglect of the notification. Some parasites transmitted from person to person can also pose a threat to public health in certain periods. Head lice, the most common examples of such parasites, can cause outbreaks in certain periods. Another example for human-induced parasitic outbreaks is scabies. There has been an increase in scabies rates in the world and in Türkiye, especially due to the Coronavirus disease-2019 (COVID-19) pandemic. In the first period of the pandemic, it was thought that due to the curfew restrictions, family members spending time at home might have led to an increase in the rate of scabies. On the other hand, as a result of the disruption of services due to COVID-19, the cases of malaria, a vector-borne disease, and the resulting deaths increased in 2020 compared to 2019 in the world. Although only imported malaria cases are detected in Türkiye today, there is a potential for an outbreak to occur at any time due to the presence of malaria vectors. An outbreak of imported malaria occurred in Mardin in 2012 due to a lorry driver entering the country from an endemic region. Immigrants that reside in Türkiye pose a risk for some infectious diseases due to the circumstances during migration or the conditions in their living areas. Leishmaniasis, which maintains its importance in the Mediterranean region, is another vector-borne disease and can be detected in Türkiye, especially in regions where immigrants reside. Bed bug infestations, which have increased recently, also closely affect the provision of health services. It is important to implement regular inspections in regions with outbreak potential, and to ensure the continuity of hygiene conditions and health services to prevent a possible outbreak. In case of an outbreak, different centers should cooperate, health authorities and academics should act together, patients and their contacts should be identified quickly and necessary precautions should be taken, the society should be informed and the outbreak should be taken under control in a short time. In this review article, outbreaks caused by parasites were examined under four headings as water, food, human and vector/arthropod-borne and examples from the world and Türkiye were given for these outbreaks.

Immune mechanisms in human Sarcoptes scabiei (Acari: Sarcoptidae) infestations.

Scabies is a parasitic infestation of human and animal skin caused by different strains of the itch mite, Sarcoptes scabiei. The World Health Organization (WHO) has declared scabies in human as a neglected tropical disease, and today over 200 million people worldwide are affected. The two most commonly reported clinical manifestation of the condition are ordinary (OS) and crusted scabies (CS). CS, which can lead to fatal consequences due to secondary bacterial infections, is mostly observed in immunocompromised subjects but can also, although rarely, be detected in immunocompetent individuals. Innate and adaptive immune system components are involved in protection and pathogenesis of scabies, although with some differences between OS and CS. While the cutaneous immune response is dominated by CD4+ T-cells in OS, it is mainly mediated by CD8+ T-cells in CS. The two clinical conditions also differ in CD4+ T-cell-mediated immune responses with mixed TH 1/TH 2 (protective) and TH 2/TH 17 (non-protective) immunoprofiles in OS and CS, respectively. Moreover, the development of CS is associated with early immunosuppression that is followed by deleterious immune response to uncontrolled mite proliferation. However, the immune response to scabies still needs further attention due to inconsistent results in the literature. The aim of this study is to attract more attention to this area by summarizing the current literature on innate and adaptive immune responses triggered against S. scabiei mites.

[Factors Influencing Pneumococcal Vaccination-Induced Total IgG, IgG2, and IgA Levels in Vaccinated Splenectomized Patients]. / Asili Splenektomili Hastalarda Pnömokok Asisi ile Indüklenmis Total IgG, IgG2 ve IgA Düzeylerini Etkileyen Faktörler.

Splenectomy is closely associated with a lifetime risk of pneumococcal and other encapsulated bacterial infections. In this study, it was aimed to investigate the change of antibody levels after vaccination against Streptococcus pneumoniae according to age, gender, years after splenectomy and the possible effect of splenectomy on IgG avidity. In addition the education and awareness levels of the participants about post-splenectomy vaccination and infectious diseases were also analyzed. In the first of the three phases of this study, 32 individuals with splenectomy were enrolled. The awareness of the patients about the possible risks after splenectomy was investigated with a simple questionnaire. Routine laboratory test results were obtained and clinical examinations were performed. In the second stage, total Ig values of 29 splenectomy patients were determined. In the third phase, 14 splenectomy and 5 healthy volunteers were vaccinated according to the Vaccination Practices Advisory Committee (ACIP) guidelines. Pneumococcal-specific antibody levels and IgG avidity were detected by enzyme linked immunosorbent assay (ELISA). It was determined that 68.8% of the splenectomized patients were unaware of their vaccination status and 78.2% of them were unaware of the increased risk of infectious diseases in asplenic conditions. . According to the hospital information management system, all 31 (96.87%) patients, except one, were vaccinated with PPV23. As expected, vaccinated patients exhibited high levels of vaccine-specific antibody production with IgG, IgG2, and IgA antibody concentrations of 321 ± 76.68 mg/l, 73.07 ± 8.273 mg/l, and 117.8 ± 14.94 mg/l, respectively, but unvaccinated patients had very low antibody (IgG, IgG2 and IgA antibody concentrations were 11.5 mg/l, 1.3 mg/l and 1.2 mg/l, respectively) levels. Although there was no correlation between antibody titers and gender, age groups or presence of fever history, the decrease in total IgG, IgG2 and IgA titers were strongly correlated with the time since splenectomy. Antibody titers were found to be significantly lower in splenectomized patients vaccinated more than 10 years ago. Routine laboratory results were at normal levels except for low platelet count. On the other hand, both splenectomized and healthy control subjects displayed similar IgG avidity index values (%61.8 ve %64.4% inhibition in control and splenectomized subjects, respectively) after the vaccination schedule. It was shown that post-splenectomy vaccination with PPV23 induced high levels of pneumococcus-specific antibody production that can last for more than five years. It was determined that more efforts should be made to increase the level of knowledge about pneumococcal and other overwhelming post-splenectomy infections (OPSI) as the awareness of the patients about the risks of infection after splenectomy was poor. In particular, patients with splenectomy operation more than 10 years ago should be very careful about being asplenic as they were determined to have significantly lower level of vaccine-specific antibody production. Our study was also the first to show that splenectomy does not alter IgG avidity induced by pneumococcal vaccination.

Serological screening of West Nile virus among blood donors in northern Cyprus.

BACKGROUND: West Nile virus (WNV) is a neurotropic arbovirus that can also be transmitted through blood transfusion. Even though its geographic distribution has been expanding, there has not yet been any epidemiological data on WNV in northern Cyprus. The aim of our study is to fill this gap by using donated blood samples. METHODS: Samples collected from the main government hospital blood bank in Nicosia were analyzed by anti-WNV enzyme-linked immunosorbent assay (ELISA) (immunoglobulin M [IgM] and immunoglobulin G [IgG]). Seropositive samples were subjected to plaque reduction neutralization test (PRNT) for confirmation and analyzed by ELISA IgG avidity test and reverse transcription real-time polymerase chain reaction (rRT-PCR). RESULTS: Of the 760 sera samples, 2 (0.3%) were IgM+ and 31 (4.1%) were IgG+. Neutralization activity was detected in none (0.0%) of the IgM+ and 26 (83.9%) of IgG+ donor specimens. ELISA IgG avidity test reported high avidity in 21 (67.7%) and low avidity in one (3.2%) IgG+ sample. PRNT-confirmed anti-WNV IgG+ samples exhibited only borderline (19.2%) or high avidity (80.8%) values. rRT-PCR results were negative for both IgM+ and IgG+ samples. CONCLUSION: Anti-WNV antibodies were detected in northern Cyprus among blood donors. The establishment of preventive measures and evaluation of the geographic extent of the WNV in northern Cyprus are highly recommended.

Thiol-disulfide homeostasis in children with celiac disease.

BACKGROUND: Toxic gliadin peptide damages enterocytes in celiac disease by causing oxidative stress. Thiols are organic compounds that defend against oxidative stress. This study aimed to investigate the changes in thiol-disulfide homeostasis in children with celiac disease. METHODS: The study included patients with celiac disease, children diagnosed with functional gastrointestinal disorders, and healthy children. Patients' serum native and total thiol-disulfide amounts, disulfide/total thiol percentage ratios, disulfide / native thiol percentage ratios, and native thiol/total thiol percentage ratios were measured. RESULTS: The study involved 172 children, of whom 90 (52.3%) were girls. The mean participant age was 8.6 ± 4.2 years. A total of 59 (34.3%) children had celiac disease, 56 (32.6%) had functional gastrointestinal disorders, and 57 (33.1%) were healthy. The total thiol and disulfide levels of patients with celiac disease (305 ± 87 µmol/L and 25 ± 15 µmol/L, respectively) were significantly lower than those of healthy children (349 ± 82 µmol/L and 40 ± 15 µmol/L, respectively) (P = 0.006 and P < 0.001, respectively). Native and total thiol levels (226 ± 85 µmol/L and 279 ± 99 µmol/L, respectively) in patients with celiac disease who consumed a gluten-containing diet were significantly lower than those of patients who consumed a gluten-free diet (278 ± 64 µmol/L and 327 ± 69 µmol/L, respectively) (P = 0.017 and P = 0.041, respectively). CONCLUSIONS: Thiol-disulfide homeostasis, an important antioxidant defense component of the gastrointestinal system, is disrupted in children with celiac disease. A gluten-free diet helped partially ameliorate this decline.

Immune mechanisms in human and canine demodicosis: A review.

Demodex mites are saprophytic parasites of the mammalian skin, mostly found in or near pilosebaceous units of hairy regions. While they can be found in healthy humans and animals without causing any clinical manifestations, they were suggested to create pathogenic symptoms when they appear in high densities under favourable conditions (ie, demodicosis). Nevertheless, their role as the primary causative agent of the pathogenic conditions in humans is debated today. Canine demodicosis, which is highly prevalent in certain dog breeds, provides a valuable tool for studying the pathogenesis of human demodicosis. Canine and human demodicosis are caused by different Demodex species, and the clinical manifestations in former could be life-threatening. Nevertheless, current literature suggests similar immune responses and immune evasion mechanisms in human and canine demodicosis; cellular immunity appeared to have a central role in protection against demodicosis, and Demodex mites were shown to influence both innate and adaptive immune response to escape immune attack. The aim of this review is to summarize the relevant literature on demodicosis obtained from studies conducted on both organisms, and draw the attention to the effect of mite-associated factors (eg, microbiota) on the different clinical manifestations displayed during human and canine demodicosis.

Skin-homing T-cell responses associated with Demodex infestation and rosacea.

AIMS: Our aim was to investigate the skin-homing T-cell immune responses triggered in patients with Demodex infestation and/or rosacea. METHODS: Collected whole blood samples were divided into four groups: control subjects; nonrosacea patients with Demodex infestation (Demodex group); papulopustular rosacea (PPR) patients without Demodex infestation (Rosacea group); and PPR patients with Demodex infestation (Rosacea/Demodex group). Following ex vivo activation, skin-homing CLA+CD4+ T-cell subset levels were monitored by flow cytometry. RESULTS: When compared with control subjects, among skin-homing CD4+ T-cell subsets analysed, Demodex patients had higher TH 9 and Treg cell levels; Rosacea subjects displayed elevated TH 1 cell levels; and Rosacea/Demodex patients exhibited increased frequencies of TH 9 and TH 22 cells. In contrast to Rosacea subjects, Rosacea/Demodex group members displayed higher TH 2 cell levels; and when compared with Demodex groups, they had higher TH 1 and TH 2 but lower Treg cell levels. Demodex group members also exhibited higher Treg but lower TH 1 and TH 22 levels than Rosacea/Demodex group subjects. CONCLUSIONS: The skin-homing T-cell responses associated with Demodex infestation and rosacea formation seem to influence each other. The present as well as future studies could contribute to the development of effective treatment strategies for demodicosis and rosacea.

Molecular identification of sulfadoxine-pyrimethamine resistance in malaria infected women who received intermittent preventive treatment in the Democratic Republic of Congo.

BACKGROUND: Point mutations in Plasmodium falciparum dihydrofolate reductase (Pfdhfr) and dihydropteroate synthase (Pfdhps) genes which confer resistance to sulfadoxine-pyrimethamine (SP) occur at increasing rates. The present study aimed to identify Pfdhfr and Pfdhps mutations in P. falciparum isolates recovered from women who received two doses of SP during pregnancy in Bandundu, the Democratic Republic of Congo (DRC). METHODS: A total of 48 women with confirmed P. falciparum infection were enrolled in the study. Finger-prick blood samples that were collected on filter paper at the time of delivery were used for DNA isolation. Pfdhfr and Pfdhps genes were amplified by a nested PCR protocol. DNA sequencing was performed on both strands, and the point mutations were analysed. RESULTS: All of the 48 (100.0%) P. falciparum isolates carried at least one polymorphism in both genes. The wild-type haplotypes of Pfdhfr (CNCSI [C50, N51, C59, S108, I164]) and Pfdhps (SAKAA [S436, A437, K540, A581, A613]) were not observed in the study. In Pfdhfr, N51I (85.4%), C59R (60.4%), and S108N (100.0%) polymorphisms were detected. Triple mutation (CIRNI) (mutant amino acids are underlined) was the most prevalent (47.9%) Pfdhfr haplotype. In the study, all P. falciparum isolates (100.0%) harboured the A437G allele in Pfdhps gene. Also, K540E and A581G polymorphisms were observed in one (2.1%) isolate. Single mutant haplotype (SGKAA) was detected in 97.9% of the isolates. Mutant Pfdhfr and Pfdhps allele combinations revealed quintuple (CICNI-SGEGA; 2.1%), quadruple (CIRNI-SGKAA; 47.9%), triple (CICNI-SGKAA; 35.4%, CNRNI-SGKAA; 12.5%), and double (CNCNI-SGKAA; 2.1%) haplotypes. CONCLUSIONS: In the study, the rate of SGEGA haplotype was low (2.1%). Although K540E and A581G alleles are more common in Eastern Africa, a distinct lineage of SGEGA is also present in the DRC, which is located in Central Africa. This haplotype is associated with decreased efficacy of SP in pregnant women and infants, therefore, it should be carefully considered in the DRC and SP resistance should be routinely monitored.

An alternative DNA extraction method for detection of Blastocystis spp. in human fecal samples.

Polymerase chain reaction (PCR) is an effective technique for diagnosis of Blastocystis infection. Notably, DNA isolation procedure is extremely critical for the PCR step. In the present study, a recently described extraction procedure, named as the "sand method" was modified and adapted for isolation of Blastocystis DNA. To evaluate its efficacy, the current method and QIAamp DNA Stool Mini Kit (Qiagen) were applied to fresh human stool samples. Our results indicated that, the mean DNA concentrations obtained by the sand method and the commercial kit were 48 and 55 ng/µl, respectively. Also, no DNA inhibitors were detected in two methods. The sand method was capable of detecting 16 parasites per 50 mg feces. DNA samples extracted by both methods were subjected to PCR. Blastocystis spp. were detected in 11 (31.4%) of 35 samples, and perfect agreement (κ: 1.000) was found between the PCR-sand method and PCR-commercial kit method. The samples that were detected positive by PCR-sand method were successfully sequenced, and Blastocystis subtypes (STs) were identified as ST3, ST2 and ST1. In conclusion, the present study indicates that the sand method provides a simple, rapid and inexpensive procedure for reliable extraction of Blastocystis DNA from stool samples.

The effect of splenectomy on the levels of PCV-13-induced memory B- and T cells.

AIM: Splenectomised patients are associated with lifelong risk of fatal overwhelming post-splenectomy infection (OPSI), which is mostly caused by Streptococcus pneumoniae. Today OPSI cases can still be reported even in patients with appropriate vaccination. In our study, the levels of vaccine-specific memory B- and T cells were compared between control and splenectomised patients to enlighten the underlying reason. MATERIALS AND METHODS: Five healthy and 14 post-traumatic splenectomised individuals were vaccinated with 13-valent pneumococcal conjugate vaccine (PCV-13) followed by 23-valent pneumococcal polysaccharide vaccine (PPV-23). The levels of memory B- and T cells were compared by ELISPOT analysis. RESULTS: Splenectomised patients generated reduced levels of memory IgG B cells in response to PCV-13 vaccination, while the memory IFN-γ T-cell levels were undetectable in asplenic patients. This was despite the detection of vaccine-induced memory T-cell levels in control patients, which were analysed simultaneously following the same experimental protocol. CONCLUSION: Our results suggest that spleen is important, but not essential, for survival and/or generation of memory IgG B cells. In contrast, it seems to be indispensable for PCV-13-specific memory TH 1-cell levels. Studies enhancing the levels of vaccine-induced memory cells and further enlightening the immune responses in asplenic individuals are required to develop more effective vaccination strategies against OPSI.

Leishmaniasis in northern Cyprus: Human cases and their association with risk factors.

BACKGROUND & OBJECTIVES: Cyprus is located in the eastern part of the Mediterranean Region where leishmaniasis is endemic. The primary objective of this study was to investigate human visceral leishmaniasis (VL) in the northern region of Cyprus where presence of canine leishmaniasis (CanL) and sandflies has been documented in earlier studies. The secondary objective was to assess the association of leishmaniasis with demographic and epidemiological variables. METHODS: Intravenous blood samples were collected from 249 volunteers in Kyrenia district (located in the northern coastal region of Cyprus). Whole blood samples were tested for DNA of Leishmania spp by polymerase chain reaction (PCR), while serum samples were analyzed using direct agglutination test (DAT) and rK39 test. For evaluation of possible risk factors, a questionnaire was applied to the participants. RESULTS: Only three (1.2%) of 249 participants were found seropositive by DAT (n = 2) or rK39 test (n = 1). The remaining samples were negative in serology, and no PCR positivity was detected in any of the 249 participants. Seven individuals, including the seropositive cases, had a history of cutaneous leishmaniasis (CL). Seropositivity and CL were not significantly related with gender (M/F: 40.2/59.8%), age [Mean: 42.85 ± 17.45, Median: 40 (7-86)], occupation (Indoor/Outdoor: 84.7/12.9%), dog ownership (52.6%), and CanL history (5.3%). However, a statistical association was found between seropositivity and past CL infection. Also, a significant relation was observed between participants living in peripheral area (63.1%) and CL infection. Furthermore, leishmaniasis awareness (28.1%) among the study population was statistically correlated with past CL infection and dog ownership. INTERPRETATION & CONCLUSION: This study demonstrates the presence of leishmaniasis and highlight the need for implementation of efficient control measures on the northern coast of Cyprus.

Cryptosporidium species and Cryptosporidium parvum subtypes in dairy calves and goat kids reared under traditional farming systems in Turkey.

Molecular characterizations of Cryptosporidium spp. in ruminants reared under traditional animal management systems are scarce and studies conducted thus far have revealed largely an absence of the pathogenic and zoonotic species Cryptosporidium parvum in pre-weaned animals. In this study, we examined Cryptosporidium species and subtype distribution in free-range pre-weaned dairy calves and goat kids with diarrhea. Cryptosporidium-positive specimens from pre-weaned calves on 10 farms and goat kids on 4 farms in Ankara, Balikesir, Corum, Kirikkale, and Kirsehir Provinces, Turkey were genotyped by PCR-restriction length polymorphism analysis of the small subunit rRNA gene, which identified C. parvum in 27 calves and 9 goat kids and Cryptosporidium ryanae in 1 calf. Among the C. parvum isolates successfully subtyped by DNA sequence analysis of the 60 kDa glycoprotein gene, three subtypes were detected in calves, including IIaA13G2R1 (20/23), IIdA18G1 (2/23), and IIdA20G1b (1/23), and four subtypes were detected in goat kids, including IIaA13G2R1 (3/8), IIaA15G1R1 (2/8), IIdA22G1 (2/8), and IIdA18G1 (1/8). Data of the study suggest that dairy calves reared in a traditional cow-calf system in Turkey are mainly infected with a C. parvum subtype rarely seen elsewhere, whereas goat kids are infected with diverse subtypes. As all five C. parvum subtypes found in this study are known human pathogens, pre-weaned farm animals could play a potential role in the transmission of human cryptosporidiosis.

Is "dried stool spots on filter paper method (DSSFP)" more sensitive and effective for detecting Blastocystis spp. and their subtypes by PCR and sequencing?

PCR and DNA sequencing are currently the diagnostic methods of choice for detection of Blastocystis spp. and their suptypes. Fresh or frozen stool samples have disadvantages in terms of several aspects such as transportation, storage, and existence of PCR inhibitors. Filter paper technology may provide a solution to these issues. The aim of the present study was to detect Blastocystis spp. and their subtypes by employing two different preservation methods: conventional frozen stool (FS) and dried stool spots on filter paper (DSSFP). Concentration and purity of DNA, sensitivity of PCR, and DNA sequencing results obtained from the two methods were also compared. A total of 230 fecal samples were included and separated into two parts: one part of the fecal samples were directly frozen and stored at -20 °C. The remaining portion of the specimens were homogenized with saline and spread onto the filter papers as thin layer with a diameter of approximately 3 cm. After air-dried, the filter papers were stored at room temperature. DSSFP samples were collected by scraping from the filter papers. DNA were extracted by EURx Stool DNA Extraction Kit from both samples. Concentration and purity were measured with Nano-Drop, then PCR and sequencing were conducted for detection of Blastocystis spp. and its genotypes. Pure DNA was obtained with a A260/A280 ratio of 1.7-2.2 in both methods. DNA yield from FS was 25-405 ng/µl and average DNA concentration was 151 ng/µl, while these were 7-339 and 122 ng/µl for DSSFP, respectively. No PCR inhibition was observed in two methods. DNA from DSSFP were found to be stable and PCR were reproducible for at least 1 year. FS-PCR- and DSSFP-PCR-positive samples were 49 (21.3 %) and 58 (25.3 %), respectively (p = 0.078). The 43 specimens were concordantly positive by both FS-PCR and DSSFP-PCR. When the microscopy was taken as the gold standard, sensitivity of DSSFP-PCR and FS-PCR was 95.5 and 86.4 %, while specificity of both tests was 99.4 and 98.3 %, respectively. DNA sequencing results of 19 microscopically confirmed cases were strictly identical (concordance 100 %) in both methods, and ST2:6, ST3:8, ST4:3, and ST6:2 were the detected subtypes. Among the 230 fecal samples, the most predominant subtypes were ST3, ST2, ST4, and ST1 by both FS and DSSFP methods. Concordance of DNA sequencing results obtained from the two methods was noted to be 90.7 %. To our knowledge, this is the first study that demonstrates DNA extraction from DSSFP is more sensitive and effective than the FS method for diagnosis of Blastocystis spp. and their subtypes by PCR and DNA sequencing.

[Evaluation of the radiological, biochemical and serological parameters of patients prediagnosed as cystic echinococcosis in Çorum, Turkey]. / Çorum'da kistik ekinokokkoz ön tanisi ile basvuran hastalarin radyolojik, biyokimyasal ve serolojik analizlerinin degerlendirilmesi.

Cystic echinococcosis (CE) is a zoonosis caused by Echinococcus granulosus. It is difficult to diagnose CE by clinical symptoms alone, therefore, radiological and serological examinations should be conducted as well. The aims of this retrospective study were to evaluate the biochemical, hemogram, serological and radiological findings of patients prediagnosed as CE, and to survey epidemiological data to detect the status of the disease in our region. A total of 253 patients (148 female, 105 male) who were admitted to Hitit University Training and Research Hospital in Corum province (located in the central Black Sea Region of Turkey), between October 2009 to July 2013, were included in the study. Serum samples collected from the patients were analyzed by indirect hemagglutination (IHA) test, in the Microbiology Reference Laboratories of the Turkish Public Health Institute, and 1/160 and higher titers were considered positive. Twenty-three (15.5%) of female patients and nine (8.6%) of male patients, with a total of 32 (12.7%) were found to be seropositive. The difference between the gender was not statistically significant (X2= 2.72). The age range of the 32 seropositive patients was between 16-90 years (mean: 51), and of them 24 (75%) being over 40 years old was found as statistically significant (X2= 22.45). All of the seropositive patients presented radiological findings diagnosed with ultrasonography and computed tomography. Additionally, it was noticed that two patients (one male, one female) who were seronegative by IHA test, have passed a CE operation and the diagnosis was confirmed with pathological findings. Of the patients 43.8% were admitted to general surgery, followed by infectious diseases (21.9%), gastroenterology (21.9%) and other (12.5%) clinics. Radiological diagnosis showed that 31 (96.9%) of seropositive patients had CE in the liver, of them two (6.3%) also had lung involvement, while one patient (3.1%) had intraperitoneal involvement alone, without liver infection. Although 50% (16/32) of patients resided in Çorum urban area, most of them were dealing with agriculture and animal breeding. Among the biochemical parameters, GGT were detected with highest level (28%), followed by ALT (16%), AST (16%) and ALP (13%), while the other parameters were normal. Elevated RDW level was the most frequently observed result (29%) among hemogram parameters, while decreased levels of hematocrit, hemoglobin and MCV were detected in 23%, 19% and 19% of the patients, respectively. Eosinophilia was detected in 19% of the patients. In conclusion, for the diagnosis of CE, which is still an important public health problem in our region, a comprehensive evaluation of clinical, radiological, serological and biochemical findings is needed, to avoid a confusion of other diseases with similar clinical symptoms.

Pediculicide regulations and usage trends in Türkiye in 2015-2022.

BACKGROUND: Over the years, the increasing prevalence of head lice infestation has led to an increase in treatment-related costs and the development of resistance to pediculicides. This study aimed to evaluate pediculicide sales over an 8-year period (2015-2022), including the COVID-19 pandemic. METHODS: Data on products registered in the Turkish Pharmaceutical Index Dataview, Intercontinental Medical Statistics (IMS) database, sold on prescription and approved by the Ministry of Health in Türkiye were formally requested from the research organization IQVIA for pharmacologically active pediculicides (PAP) such as permethrin, pyrethrin, and sumithrin. Data for physically active products containing dimethicone (DMT) were obtained from the relevant pharmaceutical companies. Sale revenues were determined based on sale prices in the aforementioned year. RESULTS: Total sales of pediculicides decreased from 2,452,846 units in 2015 to 2,028,840 units in 2022. While PAP consumption decreased over the years, from 1,832,006 to 518,460 units, DMT consumption increased from 620,840 to 1,510,380 units, from 2015 to 2022. The total price of pediculicides for the consumer increased more than threefold in Turkish currency. In terms of USD, an almost similar amount of money was spent over the years, with the exception of 2022 when the expenditure almost halved mainly due to the high inflation rate in the country. CONCLUSION: The decrease in PAP and the increase in DMT indicate that local lice have developed resistance to the former. Further studies are needed to determine the possible development of resistance in the country.

Investigation of the effects of Toxoplasma gondii on behavioral and molecular mechanism in bradyzoite stage.

Toxoplasma gondii is a single-celled parasite that causes a disease called toxoplasmosis. It can reach the central nervous system, but the mechanism of T. gondii disrupting the functioning of these brain regions occurs in bradyzoite stage of parasite, causing brain damage by forming tissue cysts in brain. In our study, the effects of T. gondii on locomotor activity, anxiety, learning and memory, and norepinephrine (NE), levodopa (L-DOPA), dopamine (DA) and 3,4-D-dihydroxyphenylacetic acid (DOPAC) catecholamines in amygdala, striatum, prefrontal cortex and hippocampus regions of the brain were investigated in bradyzoite stage. Twenty male Albino mice Mus musculus, 4-5 weeks old, weighing 20-25 g, were used. T. gondii inoculated to mice intraperitonealy with 48-50-hour passages of T. gondii RH Ankara strain. For intraperitoneal inoculation of mice 5x104 tachyzoites per mouse. No inoculation was made in control group (n: 20). Locomotor activity behavior in open field test (OFT), anxious behavior in elevated plus maze (EPM), and learning behavior in novel object recognition (NOR) tests were evaluated. NE, L-DOPA, DA and DOPAC were measured by HPLC in brain tissues of amygdala, striatum, prefrontal cortex and hippocampus. A decrease was observed in the locomotor activity, anxiety and learning values of the T. gondii group compared to the control group (p < 0.05). The heighten in NE and L-DOPA levels in amygdala tissue of T. gondii group compared to control group, an elevation in NE, L-DOPA, DA and DOPAC levels in striatum tissue, and an increase in levels of NE in prefrontal cortex tissue were detected in monoamine results. In hippocampus tissue, an increase was observed in DA levels, while a decrease was observed in NE, L-DOPA and DOPAC levels. In our study, it has been shown that T. gondii in bradyzoite stage reduces locomotor activity, causes learning and memory impairment, and has anxiogenic effects.

Clinical practice guidelines for the diagnosis and treatment of scabies.

Scabies, caused by the Sarcoptes scabiei var hominis mite burrowing into the skin, is a highly contagious disease characterized by intense nocturnal itching. Its global impact is considerable, affecting more than 200 million individuals annually and posing significant challenges to healthcare systems worldwide. Transmission occurs primarily through direct skin-to-skin contact, contributing to its widespread prevalence and emergence as a substantial public health concern affecting large populations. This review presents consensus-based clinical practice guidelines for diagnosing and managing scabies, developed through the fuzzy Delphi method by dermatology, parasitology, pediatrics, pharmacology, and public health experts. The presence of burrows containing adult female mites, their eggs, and excreta is the diagnostic hallmark of scabies. Definitive diagnosis typically involves direct microscopic examination of skin scrapings obtained from these burrows, although dermoscopy has become a diagnostic tool in clinical practice. Treatment modalities encompass topical agents, such as permethrin, balsam of Peru, precipitated sulfur, and benzyl benzoate. In cases where topical therapy proves inadequate or in instances of crusted scabies, oral ivermectin is recommended as a systemic treatment option. This comprehensive approach addresses the diagnostic and therapeutic challenges associated with scabies, optimizing patient care, and management outcomes.

Impact of wars and natural disasters on emerging and re-emerging infectious diseases.

Emerging Infectious Diseases (EIDs) and Re-Emerging Infectious Diseases (REIDs) constitute significant health problems and are becoming of major importance. Up to 75% of EIDs and REIDs have zoonotic origin. Several factors such as the destruction of natural habitats leading humans and animals to live in close proximity, ecological changes due to natural disasters, population migration resulting from war or conflict, interruption or decrease in disease prevention programs, and insufficient vector control applications and sanitation are involved in disease emergence and distribution. War and natural disasters have a great impact on the emergence/re-emergence of diseases in the population. According to a World Bank estimation, two billion people are living in poverty and fragility situations. Wars destroy health systems and infrastructure, curtail existing disease control programs, and cause population movement leading to an increase in exposure to health risks and favor the emergence of infectious diseases. A total of 432 catastrophic cases associated with natural disasters were recorded globally in 2021. Natural disasters increase the risk of EID and REID outbreaks by damaging infrastructure and leading to displacement of populations. A Generic National Action Plan covering risk assessment, mechanism for action, determination of roles and responsibilities of each sector, the establishment of a coordination mechanism, etc. should be developed.