Immunologic burden links periodontitis to acute coronary syndrome: levels of CD4 + and CD8 + T cells in gingival granulation tissue.

OBJECTIVE: To investigate the proportional variation of macrophage and T-lymphocytes subpopulations in acute coronary syndrome (ACS) patients, its association with periodontitis (P), and to compare with control individuals. SUBJECTS AND METHODS: Three groups of subjects participated: one group consisted of 17 ACS patients with P (ACS + P), another group consisted of 22 no ACS + P patients, and a control group consisted of 23 participants with gingivitis (no ACS + G). Macrophage, CD4 + , and CD8 + T-lymphocytes and CD4 + /CD8 + ratio values in gingival tissue were determined histometrically. RESULTS: Significant differences were found among three groups regarding the mean number of macrophage (no ACS + P > ACS + P > no ACS + G; p < 0.05) and CD8 + T-lymphocytes (no ACS + P > ACS + P > no ACS + G; p < 0.05). Significant variations were observed between the groups both CD4 + T-lymphocytes densities (ACS + P > no ACS + P and ACS + P > no ACS + G; p < 0.05) and CD4 + / CD8 + ratio (no ACS + P < no ACS + G and ACS + P < no ACS + G; p < 0.05). CONCLUSIONS: The increased number of CD8 + T-lymphocytes in both group ACS + P and group no ACS + P resulted in a reduction of the CD4 + /CD8 + ratio in gingival tissue when compared with no ACS + G group. CLINICAL RELEVANCE: The decrease of CD4 + /CD8 + ratio in gingival tissue reflects periodontitis and may be associated with severe adverse outcomes in people with ACS.

Comparison of a novel antigen detection test with reverse transcription polymerase chain reaction assay for laboratory diagnosis of SARS-CoV-2 infection.

Molecular diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by real-time reverse transcription polymerase chain reaction (RT-PCR) in respiratory specimens is considered the gold standard method. This method is highly sensitive and specific but it has some limitations such as being expensive and requiring special laboratory equipment and skilled personnel. RapidFor™ Antigen Rapid Test Kit is a commercially available Ag-RDT which is produced in Turkey and designed to detect the nucleocapsid antigen of SARS-CoV-2 in nasopharyngeal swab samples. The aim of this study was to evaluate the performance of this novel SARS-CoV-2 antigen detection considering the RT-PCR method as the gold standard. Four hundred forty-four nasopharyngeal swab samples which were collected from the patients who met clinical criteria of COVID-19 from ten centers in Turkey between September 2020 and February 2021 were included in the study. All the nasopharyngeal swab samples were tested for SARS-CoV-2 RNA using commercial RT-PCR kits (Bioeksen and A1 Lifesciences, Istanbul, Turkey) according to the manufacturer's instructions. Viral loads were assessed according to the cycle threshold (Ct) values. RapidFor™ SARS-CoV-2 antigen test (Vitrosens Biotechnology, Istanbul, Turkey) was used to investigate the presence of SARS-CoV-2 antigen in all samples following the manufacturer's instructions. Out of 444 nasopharyngeal swab samples tested, 346 (77.9%) were positive and 98 (22.1%) were negative for SARS-CoV-2 RNA by RTPCR. Overall sensitivity of the RapidFor™. Antigen Rapid Test Kit was 80.3% whereas specificity was found to be 87.8%. Positivity rate of rapid antigen test in samples with Ct values over 25 and below 30 was 82.7%, while it increased to 95.7% in samples 20 ≤ Ct < 25 and reached 100% in samples with Ct values below 20. RapidFor™ SARS-CoV-2 Ag test might be a good choice in the screening of symptomatic and asymptomatic patients and their contacts for taking isolation measures early, with advantages over RT-PCR as being rapid, easy and being applicable in every laboratory and even at point of care.

Efficacy and safety of an inactivated whole-virion SARS-CoV-2 vaccine (CoronaVac): interim results of a double-blind, randomised, placebo-controlled, phase 3 trial in Turkey.

BACKGROUND: CoronaVac, an inactivated whole-virion SARS-CoV-2 vaccine, has been shown to be well tolerated with a good safety profile in individuals aged 18 years and older in phase 1/2 trials, and provided a good humoral response against SARS-CoV-2. We present the interim efficacy and safety results of a phase 3 clinical trial of CoronaVac in Turkey. METHODS: This was a double-blind, randomised, placebo-controlled phase 3 trial. Volunteers aged 18-59 years with no history of COVID-19 and with negative PCR and antibody test results for SARS-CoV-2 were enrolled at 24 centres in Turkey. Exclusion criteria included (but were not limited to) immunosuppressive therapy (including steroids) within the past 6 months, bleeding disorders, asplenia, and receipt of any blood products or immunoglobulins within the past 3 months. The K1 cohort consisted of health-care workers (randomised in a 1:1 ratio), and individuals other than health-care workers were also recruited into the K2 cohort (randomised in a 2:1 ratio) using an interactive web response system. The study vaccine was 3 µg inactivated SARS-CoV-2 virion adsorbed to aluminium hydroxide in a 0·5 mL aqueous suspension. Participants received either vaccine or placebo (consisting of all vaccine components except inactivated virus) intramuscularly on days 0 and 14. The primary efficacy outcome was the prevention of PCR-confirmed symptomatic COVID-19 at least 14 days after the second dose in the per protocol population. Safety analyses were done in the intention-to-treat population. This study is registered with ClinicalTrials.gov (NCT04582344) and is active but no longer recruiting. FINDINGS: Among 11 303 volunteers screened between Sept 14, 2020, and Jan 5, 2021, 10 218 were randomly allocated. After exclusion of four participants from the vaccine group because of protocol deviations, the intention-to-treat group consisted of 10 214 participants (6646 [65·1%] in the vaccine group and 3568 [34·9%] in the placebo group) and the per protocol group consisted of 10 029 participants (6559 [65·4%] and 3470 [34·6%]) who received two doses of vaccine or placebo. During a median follow-up period of 43 days (IQR 36-48), nine cases of PCR-confirmed symptomatic COVID-19 were reported in the vaccine group (31·7 cases [14·6-59·3] per 1000 person-years) and 32 cases were reported in the placebo group (192·3 cases [135·7-261·1] per 1000 person-years) 14 days or more after the second dose, yielding a vaccine efficacy of 83·5% (95% CI 65·4-92·1; p<0·0001). The frequencies of any adverse events were 1259 (18·9%) in the vaccine group and 603 (16·9%) in the placebo group (p=0·0108) with no fatalities or grade 4 adverse events. The most common systemic adverse event was fatigue (546 [8·2%] participants in the vaccine group and 248 [7·0%] the placebo group, p=0·0228). Injection-site pain was the most frequent local adverse event (157 [2·4%] in the vaccine group and 40 [1·1%] in the placebo group, p<0·0001). INTERPRETATION: CoronaVac has high efficacy against PCR-confirmed symptomatic COVID-19 with a good safety and tolerability profile. FUNDING: Turkish Health Institutes Association.

Vaccination status among patients with the need for emergency hospitalizations related to COVID-19.

BACKGROUND: It is thought vaccines allowed for emergency use significantly reduce hospitalizations and emergency room visits. It is a matter of curiosity how many of the patients who come to the emergency department (ED) are vaccinated. We aimed to examine the characteristics of ED patients needing hospitalizations related to moderate and severe COVID-19 by vaccination status. METHODS: A retrospective study of 559 rRT-PCR-confirmed SARS-CoV-2 infection cases with moderate or severe COVID-19 needing hospitalization was performed in August 2021. Univariate and multivariate logistic regression analyses were performed for factors associated with mortality. RESULTS: The mean age of the patients was 60.8 ± 18.1 years old, and 54.2% (n = 303) of the patients were women. The most common comorbidities were hypertension (37.2%), diabetes mellitus (31.1%) and chronic obstructive pulmonary disease (13.8%), respectively. The number of patients with alpha variant was 399 (71.4%), and delta variant was 83 (14.8%). Fifty point 6% (n = 283) of the patients were fully vaccinated. The total number of patients who died in the study was 114 (20.4%), and the number of patients hospitalized in the intensive care unit was 168 (30.1%). The day between the last dose of vaccine and hospitalization was 117 ± 45.9 days. In multivariate logistic regression analysis: age (odds ratio (OR), 1.05; 95% confidence intervals (95% CI) 1.03-1.08- year increase), male gender (OR, 1.8; 95% CI, 1.1-2.9), presence of at least one comorbid disease (OR, 2; 95% CI, 1.1-3.7) and partial (OR, 0.24;95% CI, 0.09-0.6) and fully vaccinated status (OR, 0.1; 95% CI, 0.05-0.18) were associated with mortality among COVID-19 patients. CONCLUSIONS: In this study, age older than 65, unvaccinated, and comorbidities had significantly higher mortality. In multivariate regression analyses, age, vaccination status, comorbidities and the male gender were associated with mortality. Our study did not evaluate the vaccine efficacy but, a lower mortality rate was observed in those fully vaccinated with CoronaVac and Pfizer-BioNTech. Additionally, Alpha, Delta and other variants had the same mortality rates.

Severe COVID-19 and Retina: Are There Any Retinal Manifestations?

PURPOSE: To investigate whether there are retinal lesions associated with severe COVID-19. METHODS: We studied 232 symptomatic subjects aged 18 - 65 years who had severe COVID-19 and had received treatment. The evaluations included ophthalmological examinations, optical coherence tomography (OCT), imaging modalities with near infrared reflectance (NIR), fundus autofluorescence (FAF), and fundus photography (FP). RESULTS: The mean age of the patients was 49 years, and 67.6% of them were men. There were no findings of microhemorrhage, cotton wool spots (CWS), vitritis, or retinitis in the examination and imaging. CONCLUSIONS: This study indicates that retinal involvement as a complication associated with COVID-19 is questionable, although some reports have demonstrated a relationship that may occur secondary to existing systemic diseases.

Antiphospholipid antibodies in COVID-19-associated pneumonia patients in intensive care unit.

OBJECTIVES: Antiphospholipid antibodies (APAs) increase the risk of excessive blood clotting, but their role in COVID-19 remains unclear. We aimed to investigate the presence of conventional APAs used in the classification of antiphospholipid antibody syndrome in patients with severe lung infection with SARS-CoV-2 and to compare these results with non-COVID-19 critically ill patients. METHODS: Thirty-one COVID-19 patients (COVID group) and 28 non-COVID-19 critically ill patients (non-COVID group), were included in the study. Anti-cardiolipin (ACA) (IgG, IgM), anti-ß2-glycoprotein 1 (Anti-ß2GPI) (IgG, IgM, and IgA), and if the patient had not received any anti thrombotic agent before blood collection, lupus anticoagulant (LAC) tests were studied from the plasma of the patients. For testing ACA and Anti-ß2GPI, ELISA method was used, while fully automated coagulometer device was used for LAC test. RESULTS: APAs were positive in 25.81% in the COVID group (8/31) and 25% in the non-COVID group (7/28). LAC was the most common APA present in 23.08% of the COVID-19 group, who underwent measurement (6/26), while 3.57% of the non-COVID group was LAC positive (1/28) (p = .047). In the COVID group, ACA IgM, and IgG were positive in 6.45% and 0%, respectively (2/31 vs 0/31). In the non-COVID group, ACA IgM was not positive in any patient, while ACA IgG was positive in 7.14% (2/28). Anti-ß2GPI IgG and IgM tests were not positive in any patient in either the COVID or the non-COVID group. Anti-ß2GPI IgA were positive in 6.45% and 14.29%, respectively (2/31 vs 4/28). CONCLUSION: In this study, APAs were equally positive in critically ill patients among COVID-19 or non-COVID-19 patients. Only LAC was more observed in COVID-19 patients.

Remdesivir treatment for patients with moderate to severe COVID-19.

BACKGROUND: Remdesivir, which was first developed for the treatment of Ebola disease but failed to meet expectations, has become hope in the fight against the COVID-19 pandemic. This study aimed to evaluate risk factors for mortality and prognosis of adult moderate/severe COVID-19 patients treated with remdesivir, and safety and tolerability of 5 days of remdesivir treatment. METHODS: This multicenter prospective observational study was conducted in 14 centers in Turkey. Pregnancy or breastfeeding, multiorgan failure, or usage of vasopressors for septic shock, ALT > 5 × the upper limit of the normal range, or eGRF <30 mL/min or dialysis and receiving favipiravir were the exclusion criteria of the study. RESULTS: Among 500 patients, 494 patients were included in the study. On admission, 392 (79.3%) patients had moderate and 102 (20.6%) patients had severe COVID-19. The 28-day mortality was 10.1%. The median of the scores of the seven-category ordinal scale assessed on days 0, 3, 5, 7 were 4 and 3 on day 14. When the survival status of the patients was evaluated according to the time between the remdesivir start date and the end date of the symptoms, no statistically significant difference was found between the medians of the groups (p = 0.404). In multivariable analysis, age (OR, 1.05; 95%CI, 1.02-1.08; p = 0.003), SpO2 level on admission (OR, 3.03; 95%CI, 1.35-6.81; p = 0.007), heart rate (OR, 2.48; 95%CI, 1.01-6.07; p = 0.047), follow-up site at the hospital (clinic/ICU) (OR, 26.4; 95%CI, 11.6-60.17; p < 0.001) were independently associated with increased mortality. Grade 3 adverse event (AE) was observed in 4 (0.8%) patients. None of the patients experienced grade 4 or 5 AEs. DISCUSSION: Remdesivir is a safe and well-tolerated drug and older age, low SpO2 level on admission, tachycardia, and ICU admission are independently associated with increased mortality among patients with moderate/severe COVID-19 receiving remdesivir treatment.

Efficacy of therapeutic plasma exchange in patients with Crimean-Congo hemorrhagic fever.

OBJECTIVE: To examine the efficacy of therapeutic plasma exchange (TPE) in patients critically ill with Crimean-Congo hemorrhagic fever (CCHF). METHODS: Patients with CCHF received supportive treatment (ST) or TPE. After laboratory and clinical evaluations, the patients were divided into mild, moderate, and severe CCHF groups according to the severity score index (SSI). To assess the efficacy of TPE, the incubation period, time of admission to hospital, hospitalization duration, mortality rate and times to recovery of the platelet count and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were compared between patients receiving ST and TPE. RESULT: A total of 119 confirmed CCHF cases was analyzed. The median SSIs were 7 in the TPE group and 5 in the ST group. The SSI stages, median incubation times and admission times were similar in the two groups. However, the duration of hospitalization was longer in the TPE group. The overall mortality rates were 9% (3 of 33 patients) in the TPE group and 16% (5 of 31 patients) in the ST group; the difference was significant. The platelet count recovered after a median of 6 (4-7) days in the ST group. CONCLUSION: The mortality rate was lower in the TPE group than in the ST group, but the duration of hospitalization and the time to platelet recovery were longer in the TPE group than in the ST group. TPE did not contribute significantly to the prognosis of patients with intermediate-severity CCHF. However, TPE might be efficacious in patients with severe CCHF.

Low-dose tocilizumab is associated with improved outcome and a low risk of secondary infection in severe COVID-19 pneumonia.

BACKGROUND: Respiratory failure and death are the leading causes of severe Coronavirus disease 2019 (COVID-19). Hyper-inflammation and cytokine storm cause lung damage. This study aimed to compare the low-dose and high-dose effects of tocilizumab, an IL-6 receptor antagonist. METHOD: Patients with severe pneumonia and hyper-inflammation signs because of COVID-19 were included in this retrospective study. Patients receiving tocilizumab <200 mg intravenously were classified as the low-dose group, and receiving ≥200 mg as the high-dose group, and those not treated with tocilizumab as the control group. Demographic and clinical data of patients who died and survived in both low-high dose and control patients were compared. According to symptom day and radiological infiltration, patients with tocilizumab were also evaluated in two groups as early and late periods at tocilizumab administration time. RESULTS: A total of 160 patients were included in the study; 70 were treated with a low dose and 50 with high-dose tocilizumab. Forty patients were in the control group. Age, comorbidity and clinical features were similar in the control, low-dose tocilizumab and high-dose tocilizumab groups. The mortality rate (12.9%, 30.0%, 37.5, P = .008) was less in the low-dose tocilizumab group. The secondary infection rate was higher in the high-dose group than in the low-dose tocilizumab and control groups (44.0%, 10.0%, 10.0%, P < .001). Distinguishing between those patients who died and survived, age (OR: 1.1589, P < .001), higher APACHE II scores (OR: 1.225, P = .001) and needs for non-invasive mechanical ventilation (OR: 14.469, P < .001) were the most critical risk factors. Low-dose tocilizumab was associated with a lower mortality rate (OR: 0.244, P = .012). CONCLUSION: The use of tocilizumab at a low dose is associated with lower secondary infections and mortality.

From asymptomatic to critical illness: decoding various clinical stages of COVID-19

The clinical course of COVID-19 is variable, with clinical manifestation ranging from 81% mild course to 14% severe course along with 5% critical course in patients. The asymptomatic course is reported to potentially range between 20% and 70% (avg. 33%). A more severe course is seen in the elderly, those with various chronic diseases, and the immunosuppressed, where the case fatality rate is higher in these risk groups. The disease progresses with various symptoms, such as fever, cough, shortness of breath, malaise, myalgia, taste and smell disorders, diarrhea, sore throat, headache, and conjunctivitis. The disease begins with shortness of breath, indicative of lung damage, after an average of 7 to 10 days, and progresses in ARDS, sepsis, and septic shock. Some patients quickly enter shortness of breath, while others gradually develop shortness of breath and chest tightness and burning. The risk factors for a poor prognosis are age, comorbidities, and changes in laboratory tests. Secondary bacterial and fungal infections frequently develop with steroids and immunosuppressants, especially in the intensive care unit. Frequent complications in hospitalized patients include pneumonia (75%), ARDS (15%), acute renal failure (9%), and acute liver injury (19%). An increased incidence of heart damage is observed, including acute heart failure, arrhythmias, and myocarditis. Of the patients hospitalized due to COVID-19, 10%­25% present with prothrombotic coagulopathy, resulting in venous and arterial thromboembolic events. The most common extrapulmonary symptom is neuropsychiatric involvement, frequently accompanied by insomnia, an impediment to remembering, and an altered state of consciousness. During the course of COVID-19, patients undergo some pathological changes (severe lymphopenia, high levels of C-reactive protein, D-dimer, ferritin, etc.) depending on the condition and exposure level of the affected systems as shown by various laboratory tests. The relevant tests are the guiding elements of risk assessment, clinical monitoring, disease severity, and prognosis setting and therapy decision-making processes.

Emerging and reemerging respiratory viral infections up to Covid-19

Infectious diseases remain as the significant causes of human and animal morbidity and mortality, leading to extensive outbreaks and epidemics. Acute respiratory viral diseases claim over 4 million deaths and cause millions of hospitalizations in developing countries every year. Emerging viruses, especially the RNA viruses, are more pathogenic since most people have no herd immunity. The RNA viruses can adapt to the rapidly changing global and local environment due to the high error rate of their polymerases that replicate their genomes. Currently, coronavirus disease 2019 (COVID-19) is determined as an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which was first identified in 2019 in Wuhan. Herein we discuss emerging and reemerging respiratory viral infections till to SARS-CoV-2.

Investigation of anti-inflammatory effects of bee venom in experimentally induced adjuvant arthritis.

OBJECTIVES: Rheumatoid arthritis is a multisystemic inflammatory disease characterized by destruction of the joints. An effective treatment method of the disease has not been developed yet. The aim of the present study is to evaluate the effects of bee (Apis mellifera anatoliaca) venom (BV) on serum inflammatory parameters, serum antioxidant load and clinical parameters of experimentally induced adjuvant arthritis in rats. MATERIAL AND METHODS: A total of 35 Wistar albino male rats were used. The animals were divided into 5 groups. First group animals served as negative controls. The second, third, fourth and fifth groups were used for experimental arthritis induction. Following clinical development of arthritis, the first group was subcutaneously administered 0.2 ml of physiological saline, and the second, third and fourth groups were treated subcutaneously with 2 µg/kg, 4 µg/kg and 20 µg/kg once a week three times. Physiological saline injected fifth group animals were used as a sham-treatment group. Clinical observations and evaluation of arthritis were made at the 15th day, and at the end of the experiment. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, paraoxonase, serum aryl esterase, high-sensitivity C reactive protein, interleukin 1ß (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) were determined in cardiac blood samples taken at the end of the 29th day. RESULTS: From the data, total oxidant level (TOL) and oxidative stress index (OSI) were calculated. Significant improvements were observed in the clinical signs of arthritis and inflammatory markers such as in IL-1ß, TNF-α, IL-6 and TOL and OSI in the 20.0 µg/kg BV-administered group. Bee venom administration did not cause any significant increase in ALT and AST values or signs of liver toxicity. CONCLUSIONS: Bee venom treatment was effective in alleviation of symptoms of the experimental rat adjuvant arthritis by means of clinical observation and serum inflammatory markers.

Antibacterial and antibiotic-modifying activities of fractions and compounds from Albizia adianthifolia against MDR Gram-negative enteric bacteria.

BACKGROUND: Albizia adianthifolia (Schum.) is medicinally used in Cameroon to manage bronchitis and skin diseases. Our previous study documented the antibacterial potential of its roots' methanol extract. In this study, methanol roots extract was subjected to chromatography techniques and fractions (AARa and AARb), sub-fractions (AARa1-4, AARb1-2 and AARb11-14) together with isolated phytochemicals were assessed for their antimicrobial as well as their antibiotic-potentiating effects towards Gram-negative multidrug resistant (MDR) bacteria. METHODS: The antibacterial activities of the samples (determination of Minimal Inhibitory « MIC ¼ and Minimal Bactericidal Concentration « MBC ¼) were determined by the modified rapid p-iodonitrotetrazolium chloride (INT) colorimetric assay, as well as those of antibiotics in association with the compounds. Column chromatography was applied to isolate phytochemicals from roots extract and their chemical structures were determined using spectroscopic techniques. RESULTS: The phytochemicals isolated were stearic acid (1), a mixture (1:1) of stigmasterol and ß-sitosterol (2 +  3), ß-sitosterol 3-O-ß-D-glucopyranoside (4), palmatin (5), homomangiferin (6) and mangiferin (7). Fraction AARa exhibited selective inhibitory effects whilst all tested bacteria were inhibited by AARb in MIC ranges of 8 to 1024 µg/mL. Sub-fractions AARb1-2 had MIC values between 8 µg/mL and 1024 µg/mL on all tested bacteria. Phytochemicals 4, 2 +  3 and 7 inhibited the growth of 54.54% (6/11), 45.45% (5/11) and 27.27% (3/11) tested bacterial strains, respectively. When tested with an efflux pumps inhibitor (Phenylalanine-Arginine-ß-Naphthylamide or PAßN), the inhibitory effects of compounds 2 + 3 and 4 increased towards all the tested bacteria. In association with erythromycin (ERY), streptomycin (STR) and tetracycline (TET), compounds 2 + 3 and 4 had the most significant synergistic activity on the seven selected bacteria. CONCLUSION: The present study provides information on the possible use of Albizia adianthifolia and its constituents in the control of Gram-negative infections including MDR phenotypes.

Serum prolidase level in patients with brucellosis and its possible relationship with pathogenesis of the disease: a prospective observational study

Background/aim: Changes in collagen metabolism and fibroblastic activity may play a role in the pathogenesis of brucellosis. The prolidase enzyme plays an important role in collagen synthesis. We aimed to investigate the association of prolidase levels with brucellosis. Materials and methods: Serum prolidase levels in 20 patients newly diagnosed with brucellosis were compared with levels in 30 healthy control subjects. Patients with brucellosis were reassessed 3 months later for prolidase, other laboratory measurements, and response to treatment. Results: The levels of serum prolidase were significantly higher in brucellosis patients compared with those of healthy controls. Prolidase, sedimentation, and C-reactive protein levels were significantly lower after antibrucellosistreatment than before treatment. Conclusion: The current study is the first to demonstrate significantly increased serum prolidase levels in patients with brucellosis compared with healthy controls. Prolidase levels also significantly decreased with antibrucellosis treatment. This finding provides a new experimental basis to understand the pathogenesis of brucellosis in relation to collagen metabolism. The increase in serum prolidase levels might be related to several factors such as tissue destruction, increased fibroblastic activity, and granuloma formation, all of which are involved in the natural history of brucellosis.

In vitro antibacterial and antibiotic modifying activity of crude extract, fractions and 3',4',7-trihydroxyflavone from Myristica fragrans Houtt against MDR Gram-negative enteric bacteria.

BACKGROUND: Nutmeg is the seed kernel inside the fruit of Myristica fragrans Houtt. (Myristicaceae). It possesses various pharmacological activities but is used in Cameroon only for its flavor in making cakes. The present study thus aimed to investigate the in vitro antibacterial activity and antibiotic modifying activities of crude seed kernel methanol extract (MFS), fractions (MFSa-e) as well as 3',4',7-trihydroxyflavone from Myristica fragrans against a panel of multi-drug resistant (MDR) Gram-negative bacteria. METHODS: The modified rapid p-iodonitrotetrazolium chloride (INT) colorimetric assay was used to determine the Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) on the tested bacteria, as well as those of antibiotics in association with the extract and/or isolated compound. Column chromatography was used for the fractionation and purification of the seed kernel extract whilst the chemical structures of compounds were determined using spectroscopic techniques. RESULTS: Phytochemical investigations lead to the isolation of 3',4',7-trihydroxyflavone from the fraction MFSb. The crude extract showed antibacterial activity with MICs ranging from 32 to 1024 µg/mL on the majority of the 29 tested Gram-negative bacterial strains. Fraction MFSb inhibited the growth of 100% (29/29) of the tested bacterial strains, as well as the compound 3',4',7-trihydroxyflavone (12/12) with a MIC values ranging from 32 to 1024 µg/mL, and 4 to 128 µg/mL respectively. The lowest MIC value (4 µg/mL) was recorded with 3',4',7-trihydroxyflavone against Providencia stuartii ATCC299645 as well as the best MBC value (16 µg/mL) against the same strain. In the presence of Phenylalanine-Arginine-ß-Naphthylamide (PAßN), an efflux pumps inhibitor, the activity of the extract increased on 73.33% (11/15) meanwhile that of 3',4',7-trihydroxyflavone increased on 100% tested bacteria. The compound 3',4',7-trihydroxyflavone potentiated the activity of antibiotics in the majority of the tested bacterial strains. CONCLUSION: The results of the present work provide additional information on the use of nutmeg and it major antibacterial component, 3',4',7-trihydroxyflavone, as a potential drug in the treatment of bacterial infections including multidrug resistant phenotypes.

Antibacterial activities of the methanol extract, fractions and compounds from Elaeophorbia drupifera (Thonn.) Stapf. (Euphorbiaceae).

BACKGROUND: Elaeophorbia drupifera (Thonn.) Stapf. (Euphorbiaceae) is used in Cameroonian folk medicine to treat several ailments including bacterial-related diseases such as skin infections. In this study, the methanol extract from the leaves (EDL), fractions (EDLa-d), sub-fractions EDLc1-7 and EDLc31-35 as well as isolated compounds were tested for their antimicrobial activities against a panel of Gram-negative and Gram-positive bacteria including multidrug resistant (MDR) phenotypes. METHODS: The broth microdilution method was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the above samples; column chromatography was used for the fractionation and purification of the leaves extract whilst the chemical structures of compounds were determined using spectroscopic techniques. RESULTS: Phytochemical investigation lead to the isolation of a mixture (1:3) of stigmasterol and ß-sitosterol (1 + 2), euphol (3), sitosterol-O-ß- D -xylopyranoside (4), 3,3',4'-tri-O-methylellagic acid (5), a mixture (1:1) of afzelin and quercetin-3-O-ß-D-xylopyranoside (6 + 7), 3,3',4'-tri-O-methylellagic acid 4-O-ß-D-glucopyranoside (8), ellagic acid-4-O-ß-xylopyranoside-3,3',4'-trimethyl ether (9) from EDLc. Crude extract and fractions displayed selective activities with MIC values ranged from 32 to 1024 µg/mL for EDL against 84.9% of the 33 tested bacteria, 93.9% for EDLc, 69.7% for EDLb, 33.4% for EDLa and 0.03% for EDLd. MIC values ranged from 16 to 1024 µg/mL were obtained with EDLc3 and EDLc4 on all tested bacteria meanwhile other sub-fractions displayed selective activities. MIC value of 32 µg/mL was obtained with fractions EDLa against Escherichia coli AG100, EDLc against Enterobacer aerogenes ATCC13048 and EA298. For sub-fractions obtained from EDLc, the lowest MIC value of 16 µg/mL was recorded with EDLc3 against Staphylococcus aureus MRSA11. A corresponding value of 8 µg/mL against Providencia stuartii NAE16 was recorded with EDLc33 obtained from further fractionation of EDLc3. EDLc3 had MIC values below 100 µg/mL against all tested bacteria. Compound 5 as well as the mixture (1:1) of 6 and 7 inhibited the growth of all the tested bacteria with MICs ranged from 64 to 256 µg/mL. CONCLUSION: Elaeophorbia drupifera is a potential source of phytomedicine to tackle MDR bacteria. Sub-fraction EDLc3 was more active than all isolated compounds and deserves further investigations to develop natural drug to combat Gram-negative, Gram-positive bacteria and otherwise MDR phenotypes.

Evaluation of major and minor lower extremity amputation in diabetic foot patients

Background/aim: We evaluated the existing risk factors with clinical results in patients who underwent major and minor amputation of the lower extremity as a result of diabetic foot ulcers (DFUs). Materials and methods: We retrospectively studied 107 patients who had undergone lower extremity amputation. The patients were divided into minor (Group 1, n = 75) and major (Group 2, n = 32) amputation groups. On clinical evaluation, the type of surgery performed, smoking history, comorbidities, duration of diabetes mellitus (DM) diagnosis, duration of DFU presence, peripheral neuropathy, peripheral arterial disease, results of deep tissue culture, length of hospitalization, and blood parameters were investigated. Results: In Group 2, mean hospitalization time was significantly longer than in Group 1 (P < 0.05). The proportion of patients with Wagner Grade 4 was significantly higher in Group 2 than in Group 1 (P < 0.05). The duration of DM and DFU was significantly longer in Group 2 (P < 0.05). The number of polymicrobial agents was significantly higher in Group 1 (P < 0.05). Conclusion: In our study, the most important risk factors that led to major amputation in patients with DFU were age, Wagner classification, duration of DM, duration of DFU, and C-reactive protein level.

Crimean-Congo hemorrhagic fever with hyperbilirubinemia and ascites: An unusual presentation.

Crimean-Congo hemorrhagic fever (CCHF) is a fatal systemic viral infection which is an important health problem in Turkey. Many systemic symptoms have been reported including fever, hemorrhage, headache, fatigue, muscle ache, abdominal pain, nausea and vomiting. A 45-year-old male farmer with CCHF presented with massive peritoneal effusion and hyperbilirubinemia. To our knowledge, this is the first case of peritoneal effusion and hyperbilirubinemia in an adult patient with CCHF. His clinical symptoms successfully improved with supportive therapy. In patients who live in endemic areas with atypical presentation for the diagnosis of CCHF should be kept in mind.

Evaluation of the Abbott Real Time HCV genotype II assay for Hepatitis C virus genotyping.

OBJECTIVE: The determination of HCV genotypes and subtypes is very important for the selection of antiviral therapy and epidemiological studies. The aim of this study was to evaluate the performance of Abbott Real Time HCV Genotype II assay in HCV genotyping of HCV infected patients in Kayseri, Turkey. METHODS: One hundred patients with chronic hepatitis C admitted to our hospital were evaluated between June 2012 and December 2012, HCV RNA levels were determined by the COBAS® AmpliPrep/COBAS® TaqMan® 48 HCV test. HCV genotyping was investigated by the Abbott Real Time HCV Genotype II assay. With the exception of genotype 1, subtypes of HCV genotypes could not be determined by Abbott assay. Sequencing analysis was used as the reference method. RESULTS: Genotypes 1, 2, 3 and 4 were observed in 70, 4, 2 and 24 of the 100 patients, respectively, by two methods. The concordance between the two systems to determine HCV major genotypes was 100%. Of 70 patients with genotype 1, 66 showed infection with subtype 1b and 4 with subtype 1a by Abbott Real Time HCV Genotype II assay. Using sequence analysis, 61 showed infection with subtype 1b and 9 with subtype 1a. In determining of HCV genotype 1 subtypes, the difference between the two methods was not statistically significant (P>0.05). HCV genotype 4 and 3 samples were found to be subtype 4d and 3a, respectively, by sequence analysis. There were four patients with genotype 2. Sequence analysis revealed that two of these patients had type 2a and the other two had type 2b. CONCLUSION: The Abbott Real Time HCV Genotype II assay yielded results consistent with sequence analysis. However, further optimization of the Abbott Real Time HCV Genotype II assay for subtype identification of HCV is required.

Apoptosis: an underlying factor for accelerated periodontal disease associated with diabetes in rats.

OBJECTIVES: Diabetes mellitus (DM) is well-established risk factor for periodontal disease. DM can also lead to changes in the number of apoptotic cells in periodontal tissues. The goal of this study was to evaluate apoptosis, depending on DM, in healthy and diseased periodontal soft tissues. MATERIAL AND METHODS: A total of 43 adult male Sprague-Dawley rats were used in this study. Experimental periodontitis was created by placing silk ligatures around the cervices of the first mandibular molars. Experimental diabetes was induced by intraperitoneal injection of the diabetogenic agent streptozotocin (STZ). Following the induction of both experimental diseases, the animals were divided into four groups: (1) The healthy group (H) (n = 10); (2) The diabetes group (D) (n = 10); (3) The periodontitis group (P) (n = 11); and (4) The diabetes and periodontitis group (DP) (n = 12). Apoptotic cells were determined by immunohistochemistry, and the frequency of apoptotic cells was evaluated by apoptotic index score. RESULTS: It was observed that there was less apoptosis in both the epithelial and gingival connective tissue cells of healthy diabetic tissues than in healthy tissues without diabetes. When periodontal disease existed, apoptosis increased in both the epithelial and gingival connective tissues of diabetic and non-diabetic animals. CONCLUSIONS: There may be differences in the apoptotic mechanisms in the periodontal soft tissues of diabetic and non-diabetic animals. CLINICAL RELEVANCE: Apoptosis may be one of the underlying factors in increased risk for periodontal disease that is associated with diabetes.