The impact of bias of underlying literature in guidelines on its recommendations: assessment of the German fluoride guideline.

PURPOSE: The significance of the underlying literature in clinical guidelines can be weakened by the risk of bias, which could negatively affect the recommendations. Especially in controversial matters, such as fluoride use for caries prevention in children, biased results may be not reliable and lead to incorrect conclusions. This study was performed to detect bias in underlying literature of the German guideline for caries prevention using fluoride in children, where no consensus was reached between paediatricians and paediatric dentists. METHODS: Three tools used for risk of bias assessments of different study designs were RoB 2 for RCTs, ROBINS-I for non-randomized studies, and ROBIS for systematic reviews. For each study cited in the guideline two independent risk of bias assessments were performed. Disagreements were resolved by consensus. RESULTS: Out of 58 papers, 48.3% (n = 28) showed high risk of bias, with the majority in sections regarding fluoride tablets, fluoridated toothpaste, and paediatricians' recommendations. 9 out of 20 recommendations and statements were based on studies with high risk of bias, all of which were in these three controversial sections. 13 out of 29 RCTs showed high risk of bias (44.8%), as all 13 non-randomized trials did, while only 2 of 16 (12.5%) systematic reviews had high risk of bias. CONCLUSION: Considering risk of bias of cited studies in clinical guidelines may result in substantial changes in its recommendations and aid in reaching consensus. Efforts should be made to assess risk of bias of underlying literature in future clinical guidelines.

Author Correction: Impact of environmental pollutants Particulate Matter PM2.5, carbon monoxide, nitrogen dioxide and ozone on the incidence of Monkeypox cases in New York City.

Correction to: European Review for Medical and Pharmacological Sciences 2022; 26 (21): 8197-8203. DOI: 10.26355/eurrev_202211_30173-PMID: 36394769-published online on November 15, 2022. After publication, the authors applied a correction to the title: Impact of environmental pollutants Particulate Matter PM2.5, carbon monoxide, nitrogen dioxide and ozone on the incidence of Monkeypox cases There are amendments to this paper. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/30173.

Public health in Global South: effect of environmental pollutant PM2.5 on the incidence and mortality of SARS-CoV-2 in Karachi, Lahore, and Islamabad.

OBJECTIVE: Environmental pollution has undoubtedly been established as a planetary, intergenerational, and existential threat to global human health and safety. Environmental pollution is adversely affecting the world, mainly the countries where human health is not a priority aspect, and this has been exacerbated due to the Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), and pandemic is known as "COVID pandemic". This study investigates the association of environmental pollutants, particulate matter (PM2.5), with SARS-CoV-2 daily cases and deaths in Karachi, Lahore, and Islamabad, Pakistan, presenting the perspectives from the Global South. MATERIALS AND METHODS: The day-to-day PM2.5 levels were recorded from the metrological website, Real-Time Air Quality Index-AQI. The corresponding data on the COVID cases and deaths in Karachi, Lahore, and Islamabad were obtained from August 1, 2020, to September 30, 2021, from the Health Ministry and National Command Operations Centre Pakistan. RESULTS: The mean values for PM2.5 in Karachi were 110.4±46.2; in Lahore 174.0±83.2; and in Islamabad 107.1±40.0. The COVID-19 mean daily cases in Karachi were 538.9±446.6; Lahore 398.3±403.1; and Islamabad 212.2±187.6; and mean daily deaths in Karachi were 9.2±8.3; Lahore 9.3±9.7; and Islamabad 1.8±1.8. The results further depicted that the SARS-CoV-2 cases were 2.86 times higher in Karachi and 1.4 times higher in Lahore than in Islamabad. Similarly, the SARS-CoV-2 deaths were 3.6 and 2.8 times higher in Karachi and Lahore, respectively, compared to Islamabad. CONCLUSIONS: The findings claim that cases and deaths augmented significantly along with PM2.5 levels. These empirical estimates demonstrate an association between PM2.5 and SARS-CoV-2 daily cases and deaths in the cities of the Global South. These findings can contribute to policy-making decisions about addressing air pollutants and climate concerns in developing countries and create an urgency to develop a strategy for minimizing environmental pollution. This study can also steer the actions needed to address the environmental problems in developing countries to improve public health and safety.

Impact of environmental pollutants Particulate Matter PM2.5, carbon monoxide, nitrogen dioxide and ozone on the incidence of Monkeypox cases. [corrected].

OBJECTIVE: The human monkeypox disease (MPXD), is an emerging zoonotic disease caused by the monkeypox virus. The rapid spread of human monkeypox cases has developed an alarming situation worldwide. This study evaluated the impact of day-to-day air pollutants, particulate matter PM2.5, Carbon monoxide (CO), Nitrogen dioxide (NO2), and Ozone (O3) on the daily incidence of monkeypox cases in New York City, United States of America. MATERIALS AND METHODS: The daily data on air pollutants and monkeypox cases were recorded from May 1, 2022, to August 16, 2022. The everyday concentrations of "PM2.5, CO, NO2, and O3 were recorded from the metrological website "Real-Time Air Quality Index-AQI" and human monkeypox cases were documented from the official website of "NVC Health". The mean values along with correlations were performed to investigate the impact of environmental pollutants on the occurrence of monkeypox cases in New York, city USA. RESULTS: The mean value for the concentration of CO in the air was 25.61 ppm, NO2 38.16 ppm, O3 9.46 µg/m3 and PM2.5 was 1.82 ppm. The air pollutants, CO, and NO2 have a positive association (p=0.001) with daily monkeypox cases in New York, USA. The correlation analysis showed significant relationships between CO and NO2 and the number of monkeypox cases (r=0.298, p<0.002), (r=0.513, p<0.001), respectively. The linear regression analysis also showed that CO has a positive impact on monkeypox cases (ß=0.298, p<0.001). With one unit increase in the CO levels in the air, the number of monkeypox cases increased by 0.298 units, and adjusted R-square shows a 0.08 or 8% variation in the number of monkeypox cases due to an increase in CO in the environment. Moreover, NO2 has a significant positive impact on monkeypox cases (ß=0.513, p<0.001), with a one-unit increase in NO2 concentration in the air, the monkeypox cases increased by 0.513. The adjusted R-square shows that NO2 causes a 25.7% variation in the increase in monkeypox cases. However, Ozone (ß=0.018. p>0.05) and PM2.5 (ß=-0.122, p>0.05) does not have a significant correlation with monkeypox cases in the city of New York. CONCLUSIONS: Environmental pollutants NO2 and CO have a positive relationship with the number of daily monkeypox cases in New York City, USA. The air pollutants which have a high concentration in the environment have a strong relationship with the occurrence of monkeypox cases. Environmental pollution may be a risk factor for the increasing occurrence of monkeypox cases. Health officials must take priority preventive measures to curtail environmental pollution to combat the monkeypox disease.

Progranulin inhibits platelet aggregation and prolongs bleeding time in rats.

OBJECTIVE: Several adipokines secreted by adipose tissue have an anti-thrombotic and anti-atherosclerotic function. Recently identified adipokine progranulin was found to play a protective role in atherosclerosis. Bearing in mind the central role of platelets in inflammation and atherosclerosis, we aimed, in this study, to examine the effect of progranulin on platelet function and coagulation profile in rats. MATERIALS AND METHODS: Healthy male albino Wistar rats weighing (250-300 g) were divided into 4 groups. Three groups were given increasing doses of progranulin (0.001 µg, 0.01 µg, and 0.1 µg) intraperitoneally, while the control group received phosphate-buffered saline (PBS). Bleeding time, prothrombin time, activated partial thromboplastin time and platelet aggregation responses to adenosine diphosphate and arachidonic acid were assessed. RESULTS: Administration of progranulin resulted in a significant inhibition of platelet aggregation in response to both adenosine diphosphate, and arachidonic acid. Bleeding time, prothrombin time and activated partial thromboplastin time were significantly prolonged in all groups that received progranulin, in particular, the 0.1 µg dose, in comparison to the control group. CONCLUSIONS: This preliminary data is first suggesting that the antiplatelet and anticoagulant action of progranulin could have a physiological protective function against thrombotic disorders associated with obesity and atherosclerosis. However, these results merit further exploration.

The effect of superoxide dismutase enzyme inhibition on renal microcirculation of spontaneously hypertensive-stroke prone and Wistar rats.

A significant factor in the development of hypertension may be excessive vasoconstriction within the renal medulla. This study therefore investigated the role of superoxide dismutase (SOD) in the regulation of renal medullary and cortical blood perfusion (MBP and CBP, respectively) in both stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar rats. CBP and MBP were measured before and after intra-renal infusion of the SOD inhibitor, diethyldithio-carbamic acid (DETC). Under basal conditions, mean arterial pressure was significantly greater in SHRSP than Wistar rats, but both MBP and heart rate (HR) were significantly lower in SHRSP relative to Wistar rats (P<0.05, n=7 in both groups). Infusion of DETC (2 mg/kg/min) into the cortico-medullary border area of the kidney significantly decreased MBP in the SHRSPs (by 28+/-3 %, n=7, P<0.05), indicating a greater vasoconstriction within this vascular bed. However, DETC also significantly decreased MBP in Wistar rats to a similar extent (24+/-4 %, n=7, P<0.05). These results suggest that superoxide anions play a significant role in reducing renal vascular compliance within the renal medulla in both normotensive and hypertensive animals, although the responses are not greater in the hypertensive relative to the control animals.

SCH 79797, a selective PAR1 antagonist, protects against ischemia/reperfusion-induced arrhythmias in the rat hearts.

OBJECTIVE: Thrombin is implicated in the genesis of arrhythmias and activation of thrombin receptors exacerbated ventricular arrhythmias following coronary artery ligation. The present study was designed to investigate the possible protective effect of the protease-activated receptor-1 antagonist, SCH79797 against ischemia and reperfusion arrhythmias in the rat heart. MATERIALS AND METHODS: Healthy male Wistar rats (250-350 g) were anesthetized with urethane. Coronary artery ligation was performed for 5 minutes followed by 10 minutes reperfusion. Rhythm disturbances were monitored throughout the ischemia and reperfusion periods. Drugs injected were SCH79797 dihydrochloride (6.25, 12.5, 25 and 100 µg/kg), glibenclamide (5 mg/kg) and N-nitro L-arginine methyl-ester hydrochloride (25 mg/kg). The control group was injected with dimethylsulfoxide (0.1 ml). RESULTS: SCH79797 dihydrochloride reduced the number of premature contraction, prevalence and duration of ventricular tachycardia, prevalence and duration of ventricular fibrillation during both the ischemic and reperfusion periods in a dose-dependent manner. There is a trend for N-nitro L-arginine methyl-ester hydrochloride to increase all parameters of arrhythmias in SCH79797 dihydrochloride (25 µg/kg) treated rats, but glibenclamide (5 mg/kg) significantly (p < 0.05) increased these parameters. CONCLUSIONS: SCH79797 dihydrochloride induced an antiarrhythmic effect in the anesthetized rat. This protective effect could possibly be mediated by activation of nitric oxide synthase and/or of ATP-sensitive potassium channels.

Peroxiredoxin isoforms are associated with cardiovascular risk factors in type 2 diabetes mellitus

The production of oxygen free radicals in type 2 diabetes mellitus contributes to the development of complications, especially the cardiovascular-related ones. Peroxiredoxins (PRDXs) are antioxidant enzymes that combat oxidative stress. The aim of this study was to investigate the associations between the levels of PRDX isoforms (1, 2, 4, and 6) and cardiovascular risk factors in type 2 diabetes mellitus. Fifty-three patients with type 2 diabetes mellitus (28F/25M) and 25 healthy control subjects (7F/18M) were enrolled. We measured the plasma levels of each PRDX isoform and analyzed their correlations with cardiovascular risk factors. The plasma PRDX1, -2, -4, and -6 levels were higher in the diabetic patients than in the healthy control subjects. PRDX2 and -6 levels were negatively correlated with diastolic blood pressure, fasting blood sugar, and hemoglobin A1c. In contrast, PRDX1 levels were positively correlated with low-density lipoprotein and C-reactive protein levels. PRDX4 levels were negatively correlated with triglycerides. In conclusion, PRDX1, -2, -4, and -6 showed differential correlations with a variety of traditional cardiovascular risk factors. These results should encourage further research into the crosstalk between PRDX isoforms and cardiovascular risk factors.

Peroxiredoxin isoforms are associated with cardiovascular risk factors in type 2 diabetes mellitus.

The production of oxygen free radicals in type 2 diabetes mellitus contributes to the development of complications, especially the cardiovascular-related ones. Peroxiredoxins (PRDXs) are antioxidant enzymes that combat oxidative stress. The aim of this study was to investigate the associations between the levels of PRDX isoforms (1, 2, 4, and 6) and cardiovascular risk factors in type 2 diabetes mellitus. Fifty-three patients with type 2 diabetes mellitus (28F/25M) and 25 healthy control subjects (7F/18M) were enrolled. We measured the plasma levels of each PRDX isoform and analyzed their correlations with cardiovascular risk factors. The plasma PRDX1, -2, -4, and -6 levels were higher in the diabetic patients than in the healthy control subjects. PRDX2 and -6 levels were negatively correlated with diastolic blood pressure, fasting blood sugar, and hemoglobin A1c. In contrast, PRDX1 levels were positively correlated with low-density lipoprotein and C-reactive protein levels. PRDX4 levels were negatively correlated with triglycerides. In conclusion, PRDX1, -2, -4, and -6 showed differential correlations with a variety of traditional cardiovascular risk factors. These results should encourage further research into the crosstalk between PRDX isoforms and cardiovascular risk factors.

Adrenomedullin mediates early phase angiogenesis induced diabetic nephropathy in STZ diabetic rats.

OBJECTIVE: The implication of pro-angiogenic factors including vascular endothelial growth factor (VEGF) and its receptor flk-1 has been reported in diabetic nephropathy as early event. Adrenomedullin (AM), a potent vasodilator peptide, enhances angiogenesis and high levels were seen in diabetic animals and humans. However, its exact role in diabetic nephropathy is unclear. The present study investigated the effects of adrenomedullin receptor antagonist (ADM-52-22) on the early phase angiogenesis-induced diabetic nephropathy. MATERIALS AND METHODS: 28 male Wistar rats were divided into: 1) Control non-diabetic, 2) Streptozotocin (STZ)-induced diabetic rats (55 mg/kg, i.p.), 3) Control non-diabetic+ADM-52-22, and 4) STZ-diabetes+ADM-52-22 (7 per group). ADM-52-22 was infused for two weeks (250 µg/rat/day, i.p.). RESULTS: Diabetes caused an increase in kidney weight, renal VEGF levels, 24 hr urinary protein and nitric oxide excretion and hyperfiltration indicated by creatinine clearance (CrCl). ADM-22-52 reduced the rise in CrCl, total urinary protein and renal hypertrophy in diabetic rats, and attenuated early angiogenic response to diabetes: CD31 staining, flk1 protein and VEGF renal levels. CONCLUSIONS: These results show that AM through its receptor mediates early angiogenesis-induced diabetic nephropathy which attributes to the early changes as hyperfiltration and hypertrophy.

Differential associations of circulating peroxiredoxins levels with indicators of glycemic control in type 2 diabetes mellitus.

OBJECTIVES: Oxidative stress is an important patho-physiological mechanism in the development and complications of type 2 diabetes mellitus. To counteract oxidative stress the peroxiredoxin enzyme system exists in body cells. Whether diabetic state and/or glycemic control affects circulating levels of peroxiredoxins (PRDXs) needs to be elucidated. This study planned to assess PRDXs plasma levels of isoforms 1, 2, 4 and 6 in type 2 diabetes and their potential associations with glycemic control and insulin resistance. PATIENTS AND METHODS: Plasma/serum samples were obtained from type 2 diabetic patients (n=53, 28F/25M) and control non-diabetic subjects (n=25, 7F/18M). According to HbA1c diabetics were divided into well-controlled (HbA1c < 7, n=19) and poorly-controlled (HbA1c > 7, n=34). PRDXs isoforms and insulin were measured using ELISA. RESULTS: Compared to those of the control subjects, plasma levels of PRDXs1, 2, 4 and 6 were higher in diabetics. Poorly-controlled had lower levels of PRDXs2, 4 and 6 compared to well-controlled patients. PRDXs2 and 6 plasma levels negatively correlated with fasting blood sugar and HbA1c. No associations were detected with other isoforms and glycemic status or other parameters. CONCLUSIONS: The high levels of PRDXs in diabetes may suggest their chaperone function and their differential association with indicators of glycemic control may suggest their biomarker role and different mechanisms of action that warrants further investigations.

Interferon-induced Mx proteins in brain tissue of multiple sclerosis patients.

BACKGROUND AND PURPOSE: Recombinant interferon-beta is proven as an effective long-term treatment in patients with multiple sclerosis (MS). Unlike in other chronic inflammatory diseases, endogenous synthesis of type I interferons (IFN-alpha and IFN-beta) has not been studied extensively in MS. Mx proteins A and B (MxA and MxB) are intracellular proteins that are induced exclusively by type I IFNs. We investigated the expression of Mx proteins in post-mortem brain tissue of IFN-beta-naïve MS patients as a marker for endogenous synthesis of type I IFNs. METHODS: By employing monoclonal antibodies specific for MxA and MxB positive staining was detectable predominantly in reactive astrocytes within the MS plaques but also in endothelial and ependymal cells as well as in lymphocytic infiltrates. RESULTS: This is of interest in view of results previously published by our group and others that Mx protein concentrations measured by ELISA increase in blood samples from MS patients after IFN-beta therapy. CONCLUSIONS: In MS, Mx proteins are detectable in plaques suggesting endogenous synthesis of type I IFNs as part of the acute inflammatory process.

Functional Mx protein does not prevent experimental biliary atresia in Balb/c mice.

Biliary atresia (BA) is a rare cholestatic disease, which manifests itself in the form of inflammation of the liver and bile ducts in newborns, with an unknown etiology and a poor outcome. Mx proteins, which are mediators of innate, antiviral resistance induced by type I interferon, were recently detected in the livers of children with BA. Therefore, the aim of this study was to examine whether the expression of Mx protein could affect the course of experimental BA in mice. A total of 185 newborn Balb/c mice (expressing dysfunctional Mx protein) and Balb/c-Mx+-A2G mice (with functional Mx protein) were intraperitoneally infected with rhesus rotavirus (RRV) or injected with saline solution as controls. They were sacrificed if they showed signs of cholestasis or at three weeks after infection. The expression of hepatic Mx protein was detected by immunostaining (POX) and the hepatic virus load was determined. There was no significant difference in the incidence of cholestasis between wild-type Balb/c mice and Balb/c-Mx+-A2G mice (67 % vs. 65 %). However, Mx protein was highly expressed in Balb/c-Mx+-A2G mice with BA phenotype, but not in wild type Balb/c mice or disease-free Balb/c-Mx+-A2G mice despite RRV infection. The difference in the hepatic virus load was not statistically significant in mice with BA. In conclusion, Mx protein does not prevent newborn Balb/c mice from developing biliary atresia after RRV infection. However, the expression of Mx protein is independent of the hepatic virus load and could be used as a marker of BA in humans, as well as in the RRV model.

Mx proteins in blood leukocytes for monitoring interferon beta-1b therapy in patients with MS.

OBJECTIVE: To correlate Mx protein (Mx) levels in lysed blood leukocytes with the clinical response to interferon (IFN) beta-1b (IFNbeta-1b) in relapsing-remitting MS (RR-MS) patients for monitoring treatment. BACKGROUND: Intracellular Mx expression is exclusively induced by the type I IFNs (IFN-alpha, -beta, and -omega) or by viruses and is strongly increased under IFN treatment. Quantitative determination of Mx allows objective assessment of biological effects of IFN. METHODS: Mx protein levels were measured in blood leukocyte lysates from IFNbeta-1b-treated RR-MS patients by ELISA and correlated to clinical parameters, including relapse rate and clinical deterioration. RESULTS: In stable IFNbeta-1b-treated MS patients, Mx levels were significantly increased compared to patients with or without immunosuppressive treatment. In IFN-1b-treated MS patients during relapse, Mx levels were significantly lower than during stable phases of the disease. Mean values of Mx (MVMx) over time of treatment in patients with a reduction of relapse rate were significantly higher than in patients without response. CONCLUSION: Mx levels in lysed blood cells may represent a useful surrogate marker for IFNbeta-1b activity corresponding to the clinical response during treatment of MS.

Expression of interferon-inducible Mx-proteins in patients with IgA nephropathy or Henoch-Schönlein purpura.

Both viral infections and dysregulated cytokine synthesis have been implicated in the pathogenesis of immunoglobulin A nephropathy (IgAN) and Henoch-Schönlein purpura (HSP). Mx proteins are specifically induced by type I interferons (IFN-alpha, -beta, -omega) and are very sensitive in detecting, for example, virus-induced, in vivo production of IFN-alpha/-beta, because the biological half-life of Mx (approximately 3 days) markedly exceeds that of IFN-alpha/-beta (20 to 90 minutes). Mx concentrations in leukocytes were measured by enzyme-linked immunosorbent assay (ELISA) in 79 blood samples of 35 patients with IgAN and five with HSP. No patient showed symptoms of infections at the time of the examination. Compared with normal leukocyte Mx concentrations (<2 mU/1,000 leukocytes), only 3 of 79 samples of IgAN/HSP patients showed mildly elevated Mx concentrations (range, 2.2 to 3 mU/1,000 leukocytes). By contrast, patients with increased endogenous IFN production (lupus erythematosus) or patients treated with IFN-alpha2 showed leukocyte Mx concentrations of up to 35 mU/1,000 leukocytes. In patients with IgAN and HSP, leukocyte Mx concentrations were not correlated with various clinical parameters. Immunohistochemically, no renal Mx expression could be detected in eight renal biopsy specimens of patients with various stages of IgAN, whereas control specimens (skin of patients treated with IFN-alpha2) showed abundant cellular Mx expression. Furthermore, human mesangial cells in vitro showed marked Mx production after exposure to IFN-alpha or IFN-beta. We conclude that, in patients with IgAN/HSP, no evidence of an activation or dysregulation of the type I interferon system can be detected.

Intracellular staining of Mx proteins in cells from peripheral blood, bone marrow and skin.

AIM/BACKGROUND: The Mx proteins are known to be specifically and dose dependently induced in mononuclear cells (MNC) by type I interferons (IFN). The aim of this study was to establish a staining method for the human intracellular Mx proteins, MxA and MxB, in leucocytes and bone marrow and skin cells. METHODS: Several monoclonal antibodies directed against the MxA and MxB proteins were generated. These antibodies were used to stain Mx proteins in both frozen and paraffin wax sections using the standard alkaline phosphatase anti-alkaline phosphatase (APAAP) method. RESULTS: Granulocytes, monocytes and lymphocytes extracted from freshly collected blood from 21 healthy subjects did not stain. After incubating MNC from these subjects with IFN alpha 2b for 48 hours, Mx proteins were detected in monocytes and lymphocytes. Within two days of starting treatment with subcutaneous IFN alpha 2b, granulocytes, monocytes and lymphocytes of 16 patients with cancer stained strongly for Mx proteins. The intensity of staining was correlated with the Mx content of whole blood measured using a specific ELISA. Prior to IFN treatment, cells from bone marrow and skin tissue specimens were negative for Mx proteins with the exception of endothelial cells. During treatment with IFN alpha 2b, nearly all cells from bone marrow and skin stained intensely. CONCLUSIONS: These new monoclonal antibodies facilitate the detection of Mx positive cells in peripheral blood and in frozen or paraffin wax specimens. The advantage of this staining method is that individual cells which have responded to viruses or biologically active IFN alpha, beta or omega can be identified.