Mitochondrial Fraction of Circulating Cell-Free DNA as an Indicator of Human Pathology.

Circulating cell-free DNA (ccfDNA) of mitochondrial origin (ccf-mtDNA) consists of a minor fraction of total ccfDNA in blood or in other biological fluids. Aberrant levels of ccf-mtDNA have been observed in many pathologies. Here, we introduce a simple and effective standardized Taqman probe-based dual-qPCR assay for the simultaneous detection and relative quantification of nuclear and mitochondrial fragments of ccfDNA. Three pathologies of major burden, one malignancy (Breast Cancer, BrCa), one inflammatory (Osteoarthritis, OA) and one metabolic (Type 2 Diabetes, T2D), were studied. Higher levels of ccf-mtDNA were detected both in BrCa and T2D in relation to health, but not in OA. In BrCa, hormonal receptor status was associated with ccf-mtDNA levels. Machine learning analysis of ccf-mtDNA datasets was used to build biosignatures of clinical relevance. (A) a three-feature biosignature discriminating between health and BrCa (AUC: 0.887) and a five-feature biosignature for predicting the overall survival of BrCa patients (Concordance Index: 0.756). (B) a five-feature biosignature stratifying among T2D, prediabetes and health (AUC: 0.772); a five-feature biosignature discriminating between T2D and health (AUC: 0.797); and a four-feature biosignature identifying prediabetes from health (AUC: 0.795). (C) a biosignature including total plasma ccfDNA with very high performance in discriminating OA from health (AUC: 0.934). Aberrant ccf-mtDNA levels could have diagnostic/prognostic potential in BrCa and Diabetes, while the developed multiparameter biosignatures can add value to their clinical management.

Total IgE, eosinophils, and interleukins 16, 17A, and 23 correlations in severe bullous pemphigoid and treatment implications.

Bullous pemphigoid (BP) patients are predominantly above 70 years of age, with limited tolerance to the side effects of the immunosuppressive drugs. Advancements in our understanding of the pathophysiology of BP have led to the development of molecules which target specific pathways involved in induction and perpetuation of disease. Patients with BP Disease Area Index above 60 and less than 100 were split into two groups-one with high and the other with normal levels of IgE. The tested parameters included eosinophils' count, total IgE serum level, and interleukins (IL) 16, 17A, and 23 counts in the peripheral blood and skin bulla serum, before any therapeutic intervention. Thirty individuals fulfilled the criteria for enrollment. Patients with high IgE blood serum levels had significantly higher levels of IL17A and normal IL23 levels in blood and bulla serum. Patients with normal serum IgE levels had slightly higher IL23 levels in blood and bulla serum. The eosinophil count was positively related to IL17 blood serum level and negatively related to IL23. IL16 did not differ in the two groups. BP patients may represent a group of patients benefiting most substantially from the introduction of nonimmunosuppressive therapeutics into the treatment regimens for their disease. Clinical criteria and immune biomarkers are needed for making the best therapeutic choice.

Relative Adrenal Insufficiency is Associated with the Clinical Outcome in Patients with Stable Decompensated Cirrhosis.

BACKGROUND: The clinical impact of relative adrenal insufficiency (AI) on patients with stable decompensated cirrhosis (DeCi) has not been yet elucidated. AIM: Explore the association between AI and outcome [death or liver transplantation (LT)] in patients with DeCi. MATERIAL AND METHODS: Patients with DeCi presenting no active complication have been included. Clinical and laboratory data, including serum levels of corticosteroid-binding globulin (CBG), interleukin (IL)-1b, IL-6 and tumor necrosis factor (TNFα) were recorded in each participant. Salivary cortisol (SC) and serum total cortisol (STC) were assessed at (T0) and 1 h (T60) after intravenous injection of 250 µg corticotropin. RESULTS: 113 consecutive patients were totally tested. Median SC was 3.9 ng/mL and 15.5 ng/mL and median STC was 10.7 µg/dL and 22.7 µg/dL at T0 and T60 respectively. The patients with AI [group 1, n = 34 (30%)] had significantly lower systolic blood pressure (106 ± 12 vs. 113 ± 13 mmHg, p = 0.05), serum sodium (133 ± 7 vs. 137 ± 12 mEq/ L, p = 0.04), HDL (29.9 ± 14 vs. 38.6 ± 18 mg/dL, p = 0.034) and albumin (2.7 ± 0.5 vs. 3.1 ± 0.5 g/dL, p = 0.002), but higher direct bilirubin (median: 1.6 vs. 0.8 mg/dL, p = 0.029) compared to those without AI [group 2, n = 79 (70%)]. Moreover, group 1 patients presented more frequently past history of spontaneous bacterial peritonitis (SBP) [10/34 (29.4%) vs. 6/79 (7.5%), p = 0.002]. AI was significantly associated with death [HR = 2.65, 95% C.I.: 1.55 - 4.52, p = 0.003 over a follow up period of 12 (6-48) months.] Conclusions. The presence of AI in patients with stable DeCi predispose to obvious clinical implications since it is associated with circulatory dysfunction, previous history of SBP and worse survival.

Syncope without prodromes is associated with excessive plasma release of adenosine at the time of syncope during head-up tilt table test.

BACKGROUND: In syncopal patients without underlying structural disease, we sought to investigate the association of Adenosine Plasma Levels (ADP) with the clinical presentation of neurally mediated syncope (NMS) and the outcomes of Head-Up Tilt Table Test (HUTT) and Adenosine test (ADT). METHODS: We studied 124 patients with different clinical types of NMS, i.e., Vasovagal (VVS, n=58), non-prodromes (NPS, n=18), or situational syncope (SS, n=48), using a standard protocol including HUTT and ADT. During HUTT, ADP was measured in the supine position, at table tilting and in syncope. RESULTS: Baseline ADP did not differ among groups. ADP at syncope were higher in NPS (n=5) compared to VVS (n=20): 0.23 vs. 0.12 µΜ, p=0.03, and SS (n=22): 0.04 µΜ, p=0.02. In NPS, ADP increased from supine to syncope (n=5): 0.15 vs. 0.23 µΜ, p=0.04. In VVS, ADP increased only from supine to tilt position: 0.11 vs. 0.14 µΜ, p=0.02. In SS, ADP did not change during HUTT. In positive vasodepressor HUTT, ADP increased from supine to tilt position (p=0.002) and at syncope (p=0.01). In SS, 20.0% exhibited cardioinhibitory HUTT vs. 6.8% in other forms of syncope (p=0.04). In SS, 22.9% manifested positive ADT vs 6.6% in other types of syncope (p=0.012). CONCLUSION: The subset of NPS patients with positive HUTT, show excessive ADP release at the time of syncope. This may explain the lack of prodromes in this form of syncope. Such observations contribute to the understanding of distinct profiles of clinical forms of syncope and may differentiate the management approach accordingly.

Total antioxidant status in patients with obstructive sleep apnea without comorbidities: the role of the severity of the disease.

OBJECTIVE: Several studies suggest that obstructive sleep apnea syndrome (OSAS) is associated with oxidative stress. However, there are also contrary observations and the role of antioxidant mechanisms has not been fully explored. PATIENTS AND METHODS: The present study evaluated serum total antioxidant status (TAS) in 32 OSAS patients without comorbidities, before and after a diagnostic sleep study and at a second sleep study after continuous positive airway pressure (CPAP) application. RESULTS: Lower TAS values were found in the morning, immediately after the first sleep study, compared with those before sleep (1.68 ± 0.11 vs. 1.61 ± 0.10 mmol/l, p < 0.01); this was evident in patients with less severe syndrome [apnea-hypopnea index (AHI) <30; 1.73 ± 0.08 vs. 1.65 ± 0.09 mmol/l, p = 0.01] but not in severe syndrome (AHI ≥30; 1.64 ± 0.12 vs. 1.58 ± 0.10 mmol/l, p = 0.07). After CPAP application, a statistically significant decrease in TAS values was observed in patients with less severe syndrome (1.72 ± 0.05 vs. 1.63 ± 0.04 mmol/l, p = 0.003). On the contrary, no statistically significant changes in TAS were observed in patients with severe syndrome. CONCLUSIONS: The present study supports a reduction in antioxidant capacity during sleep in otherwise healthy patients with OSAS. This reduction was evident in less severe syndrome. However, a single night of CPAP application seems to ameliorate this antioxidant disturbance only in less severe syndrome.

Cerebral folate deficiency in two siblings caused by biallelic variants including a novel mutation of FOLR1 gene: Intrafamilial heterogeneity following early treatment and the role of ketogenic diet.

Mutations in the FOLR1 gene, encoding for the folate alpha receptor (FRa), represent a rare recessive genetic cause of cerebral folate deficiency (CFD), a potentially reversible neurometabolic condition. Patients typically present with developmental delay, seizures, abnormal movements, and delayed myelination. We hereby expand the phenotypic and genotypic spectrum of the disease with the report of the first two Greek siblings that were found compound heterozygous for one known FOLR1 gene mutation (p.Cys65Trp) and a mutation (p.Trp143Arg) that has not yet been reported in the literature (class 3 variant according to ASHG classification). A distinguishing feature of the older sibling is the manifestation of drug-resistant epileptic spasms beyond infancy. These had a relatively good response to a ketogenic diet, as an additional treatment to topiramate and valproate. A further clinical improvement was observed when folinic acid was combined with the above treatment. While a response to folinic acid is well established in the disorder, the efficacy of its combination with the ketogenic diet needs further evaluation, but we suggest considering it early in the course of drug resistant epilepsy in the setting of CFD. The younger sibling was diagnosed and treated with folinic acid at an early-symptomatic stage. Both patients had moderately low age-related CSF 5-methyltetrahydrofolate levels at diagnosis with the older sibling (that was already treated at base line collection) averaging 19 nmol/L (normal range: 44-122 nmol/L) and the younger one 49 nmol/L (normal range 63-122 nmol/L). These levels were restored to normal limits after folinic supplementation.

Genetic study of the distribution of Greek goat encephalitis virus in Greece.

Tick-borne encephalitis virus (TBEV) and Louping ill virus (LIV) are viruses in the Mammalian tick-borne virus group/genus Flavivirus, causing central nervous system disease. Greek goat encephalitis virus (GGEV), which was isolated from the brain of a newborn goat with neurological symptoms, is currently classified in the TBEV group. The vector of GGEV has not yet been specifically identified but is considered likely to be Ixodes ricinus. A total of 3,144 Ixodidae ticks collected during 2003-2006 from goats and sheep in rural areas of Northern Greece were tested by reverse transcription-polymerase chain reaction (RT-PCR) for the presence of TBEV-specific RNA. Two pools of I. ricinus ticks were shown to be RNA-positive. Sequence analysis showed that the virus was GGEV. The RNA-positive ticks were detected in regions where high prevalence of TBE antibodies in humans was present. Prevalence in ticks varied according to year, season, and geographic region. TBEV is not endemic in Greece, and most probably the seroprevalence of TBE antibodies in humans is due to cross-reactivity to GGEV.

Ticks infesting domestic animals in northern Greece.

The tick species infesting grazing animals in the countryside of 11 prefectures in Northern Greece were investigated during April-July and September-December of consecutive years 2003-2006. A total of 3,249 (1,952 males, 1,297 females) adult ticks were collected from goats, sheep, cattle and dogs. Ticks were identified as Ixodes ricinus (44.57%), Ixodes gibbosus (4.09%), Rhipicephalus bursa (19.14%), Rhipicephalus sanguineus, Rhipicephalus turanicus (5.79%), Hyalomma marginatum marginatum (12.40%), Dermacentor marginatus (0.31%) and Boophilus annulatus (4.43%). Rhipicephalus spp. and Hyalomma spp. were abundant in all prefectures, Ixodes spp. were present in 9/11 prefectures, Boophilus spp. in 4/11, while Dermacentor spp. were found only in one. Results of this study give an insight into the ecology of ticks and their potential of tick-borne diseases in the country.

The epidemic West Nile virus strain in Greece was a recent introduction.

To estimate the prevalence of West Nile virus (WNV) immunoglobulin G (IgG) antibodies in residents of northern Greece before the large outbreak of 2010, stored serum samples collected during 2003-2004 from 626 apparently healthy residents of northern Greece were tested. A seroprevalence of 0.62% was detected, with no statistically significant differences among the various prefectures, gender, and age groups, suggesting that the lineage 2 WNV strain that caused the outbreaks for 3 consecutive years was introduced recently. Data from the present study can be used as a threshold in future comparisons with respective data after the outbreaks to estimate and assess the public health impact, as well as for WNV evolutionary studies.

Acute adrenal insufficiency following arthroplasty: a case report and review of the literature.

BACKGROUND: Acute adrenal insufficiency is a potentially lethal condition rarely caused by bilateral adrenal haemorrhage due to heparin use. Most of the times, it is difficult to establish the diagnosis, as symptoms are not specific. Few cases have been reported in the literature. CASE PRESENTATION: A 52-year-old Caucasian woman presented with abdominal pain, vomiting and weakness nine days after arthroplasty and heparin use. Hyperkalemia, low cortisol and high adrenocorticotropic hormone levels were found, indicating adrenal insufficiency. Magnetic resonance imaging of the upper abdomen was compatible with preceding adrenal haemorrhage. Hydrocortisone and fludrocortisone were administered. Review of the literature revealed 36 cases of postoperative adrenal haemorrhage which are presented briefly. CONCLUSION: Postoperative acute adrenal insufficiency due to haemorrhage is a rare condition. If patients are treated based on clinical suspicion, they have good chances to survive. Hydrocortisone is given permanently in the majority of the patients.