Volume 28 (2020): Issue 2 (April 2020)

Multiplex Ligation-dependent Probe Amplification is a technique proposed for the detection of deletions or duplications that may lead to copy number variations in genomic DNA, mainly due to its higher resolution, and shorter overall diagnosis time, when compared with techniques traditionally used, namely karyotyping, fluorescence in situ hybridization, and array comparative genomic hybridization. Multiplex Ligation-dependent Probe Amplification is a fast (about 2 days), useful and cost-effective technique, being suitable for the diagnosis of hereditary conditions caused by complete or partial gene deletions or duplications, as these conditions are either more difficult or impossible to be diagnosed by other techniques, such as PCR, Real-Time PCR, or sequencing (Sanger or Next Generation). Due to its numerous advantages over conventional cytogenetic analysis techniques, Multiplex Ligation-dependent Probe Amplification could be used in the near future as the main technique for the molecular investigation of genetic conditions caused by copy number variations, in both rare and complex genetic disorders.

Acute lymphoblastic leukemia is the most common hematological malignancy at pediatric age. Cardiotoxicity holds the first place among the causes of morbidity and mortality in these patients. Anthracyclines are cytostatic drugs frequently associated with cardiotoxicity. Early diagnosis of cardiac impairment during the treatment of pediatric patients is extremely important, both for modulating future chemotherapy and for administering cardioprotective agents. Long term monitoring after chemotherapy helps to identify the risk of late cardiotoxicity among cancer survivors. There are several biomarkers, already in use or still under study, which may represent an operator-independent alternative for echocardiography in the diagnosis of cardiotoxicity. In case of cardiac damage, the clinician has options for treating or limiting the progression, either with the use of already approved agents, such as Dexrazoxane, or by administrating other cardioprotective drugs. International experts are still attempting to establish the best algorithm for early detection of cardiotoxicity, as well as the most efficient treatment plan in case of already existing myocardial damage in these patients. We present a review on treatment-related cardiotoxicity, including mechanisms of development, useful biomarkers and treatment options, after carefully analyzing specialty literature.

Aim: The aim of this study is to identify correlations between total antioxidant status values of mothers and their infants and compare these values in accordance to the presence or absence of intrauterine growth restriction.

Material and methods: This is a prospective, comparative study performed over a period of 3 years on a number of 52 infants and their mothers. Thirty-six of them had intrauterine growth restriction and 16 were appropriate for their gestational age and were used for comparative purposes. General information regarding the mother, infant and pregnancy were recorded. In addition, total antioxidant status was assessed from blood samples, taken right before delivery from mothers and from the cord blood in infants.

Results: We found significant differences between total antioxidant status both between mothers and neonates with IUGR (intrauterine growth restriction) versus without IUGR (p=0.018, and p<0.001, respectively). In addition, in both groups, there was a significant direct correlation between maternal and neonatal values of serum total antioxidant status (TAS) (p<0.001).

In conclusion, we can say that TAS values, as an important marker of the oxidative status of patients, are correlated with the presence of IUGR and values recorded from blood samples of the mother may be predictive for the oxidative status of the infant, thus of IUGR.

Background: Despite the fact that keratoconus has been tipically defined as a noninflammatory condition, recent research has promoted the role of inflammatory factors and protein changes of tear film in disease progression.

Aims: to determine the level of serum albumin, lactoferrin and lyzozyme in tears of keratoconic patients and their correlations with corneal biomechanical properties.

Subjects and methods: 16 eyes of keratoconus patients and 14 eyes of control cases were enrolled in an observational prospective study. We performed a complete ophthalmological examination on all participants. In order to determine the concentration of tear film proteins, a minimum of 20 microlitres of tears from the lower conjunctival fornix were collected from each subject and measured by enzyme-linked immunosorbent assay (ELISA) analysis.

Results: The level of lactoferrin measured in the tear film was significantly decreased in the keratoconus group compared to the normal subjects in all cases (p<0.05). We also found an increased level of lyzozyme and albumin in the keratoconus patients when compared to the controls, only the lyzozyme beeing statistically significant. In the keratoconus group, the correlations between proteins and important parameters such as keratometry, pachymetry and corneal biomechanics were statistically relevant in our study.

Conclusions: We can state that the protein composition of tears is modified in keratoconus by increased levels of protein with inflammatory properties such as albumin or by decreased levels of protein with anti-inflammatory properties such as lactoferrin.

Background: Conflictual results regarding the relationship between plasmatic level of visfatin and obesity could be explained by the influence of the gene variants involved in the synthesis or action of these hormones.

Objectives: The present study examined the potential implication of single nucleotide polymorphisms (SNPs) of nicotinamide phosphoribosyltransferase (NAMPT) gene that encodes visfatin, in obesity, in a Romanian pediatric population.

Method: A case-control study was conducted on a group of 213 children, divided into two: the case group - 130 overweight and obese children with BMI >1 SD, and the control group - 83 children with normal BMI. The variables analyzed were age, sex, anthropometric parameters, body composition based on bioimpedance analysis, lipid profile, visfatin and insulin plasmatic levels, rs4730153 and rs2302559 visfatin SNPs.

Results: Significant associations were not found between rs4730153 and rs2302559 visfatin SNPs and obesity. Regarding lipid metabolism, there are statistically significant differences between triglyceride levels according to NAMPT rs2302559 genotypes (p=0.045), with heterozygous genotype having the highest level of triglycerides, and also between cholesterol levels according to NAMPT rs4730153 genotypes (p=0.030), with carriers of heterozygote genotype having the highest level of cholesterol. There is a statistically significant difference between the studied parameters in the investigated groups, regarding cholesterol, in carrier of wild-type genotype of NAMPT rs2302559 (p=0.040) and carrier of wild-type genotype of NAMPT rs4730153 (p=0.036). We observed no association of NAMPT rs4730153 and rs2302559 with visfatin levels in the studied groups. Visfatin level was lower in the case group and was correlated with weight (p=0.042), abdominal circumference (p=0.010), waist to height ratio (p=0.040), but not with the elements of the metabolic profile.

Conclusion: NAMPT rs2302559 and rs4730153 polymorphisms do not seem to have a major impact in the development of obesity in children, however there may be an association with lipid profile, but further studies are needed..

To date it is unknown if there is a predisposition to sepsis. In this respect, genetic studies have been conducted with the aim to find gene variants which can point out a higher predisposition to developing sepsis. The primary objective of this study is to highlight whether the genetic polymorphism of Angiopoietin-2 gene (ANG2-35G>C) is present mainly in septic patients. As secondary objectives we aimed to evaluate if there are any associations between ANG2-35G>C polymorphism and the severity scores Acute Physiology and Chronic Health Evaluation II (APACHE II) and Simplified Acute Physiology Score (SAPS) as well as routine tests in septic patients such as C reactive protein (CRP), procalcitonin (PCT). We enrolled adult patients admitted to the Intensive Care Unit (ICU). After admission to the ICU and the diagnosis of sepsis, blood samples were collected and the severity scores: APACHE II, SAPS were calculated on the first day of ICU admission. We recorded the following from the blood samples: CRP, PCT, angiopoietine2 (Ang-2). We performed several one-way ANOVA tests to determine any significant mean difference of the analyzed variables. We observed that variant genotypes of ANG2-35G>C gene polymorphism are significantly related to CRP, aspect which increases this biomarker credibility compared with others (i.e., PCT), in septic patients. ANG2-35G>C gene polymorphism is associated with severity scores, APACHE II, and SAPS in sepsis.

Introduction: Our study investigated the importance of inflammation markers – ratio of platelets and lymphocytes (PLR), ratio of neutrophils and lymphocytes (NLR) and ratio of lymphocytes and monocytes (LMR) – as predictive markers in the occurrence of fistula or stenosis in patients diagnosed with gastric adenocarcinoma who underwent gastric resections.

Materials and Methods: We conducted a retrospective study of 178 patients diagnosed with gastric adenocarcinoma. The included patients were divided into 3 groups: group 1 (77 patients, who underwent lower gastrectomy), group 2 (27 patients, who had upper polar gastrectomy otherwise known as proximal gastrectomy), group 3 (74 patients, who underwent total gastrectomy). Ratios of PLR, NLR, respectively LMR were calculated for all patients.

Results: Out of 178 patients 52 (29.2%) developed postoperative stenosis and 16 patients (9.0%) had postoperative fistulae. The occurrence of anastomotic stenosis was associated with significantly higher preoperative platelet counts (p=0.043) and PLR values (p=0.023). ROC curve analysis indicated that the optimal PLR value for the prediction of gastric stenosis was 198.4 (AUC= 0.609, sensitivity: 59.6%, specificity: 61.9%). For the prediction of fistulization PRL also displayed the highest performance among the analyzed hematological parameters (AUC=0.561, sensitivity: 43.7%, specificity: 81.5%, cut-off value 116.6.

Conclusion: Our study indicates the importance of PLR as e predictive factor in the occurrence of anastomotic complications (fistulae or stenosis) immediately following surgery in patients with gastric adenocarcinoma that undergo gastric resections. Further prospective studies on larger groups of patients are required, considering that PLR, NLR and LMR will be key markers in the clinical management of patients with gastric cancer.

Aim: We aimed to examine the association between several circulating bone turnover markers [ osteocalcin (OC), osteoprotegerin (OPG), beta-CrossLaps (β-CTx)], hip and spine bone mineral density (BMD) and abdominal aortic calcification (AAC) in patients with chondrocalcinosis (CC).

Methods: Thirty-six patients with CC and thirty-seven controls were consecutively enrolled in this pilot case-control, cross-sectional study. The following parameters were assessed: serum levels of OC, OPG and β-CTx by enzyme-linked immunosorbent assay (ELISA); hip and spine BMD by dual-energy X-ray absorptiometry and AAC score by lateral radiography.

Results: Patients with CC had higher levels of serum bone turnover markers and AAC score than the control group: OC [6.5 (3.5-9.9) vs 4.5 (2.6-7.2) ng/ml; p=0.05], OPG [(7.7 (6.2-9.4) vs 6.5 (5.5-8.12) pmol/ml; p=0.02], β-CTx [6078 (5870-6171) vs 5851 (5465-6109) pg/ml; p=0.02] and AAC score (3.6±6.2 vs 0.5±2; p=0.006). Conversely, even if statistical significance was not reached, hip and spine BMD was lower in patients with CC. Additionally, we found a positive correlation between OPG and AAC, but also between OPG and osteoporosis in patients with CC.

Conclusion: Patients with CC are characterized by higher circulating OC, OPG and β-CTx. The presence of AAC was more common in patients with CC, being only associated with serum OPG.

The increasing resistance against classical antibiotic treatment forces the researchers to develop novel non-toxic antimicrobial agents. The aim of this study was to determine the antimicrobial properties of seven different porphyrins having distinctive hydrophobicity/hydrophilicity: P1 meso-tetra(4-methoxy-phenyl)porphyrin, P2 Zn(II)-meso-5,10,15,20-tetrapyridylporphyrin, P3 meso-tetra(p-tolyl)porphyrin, P4 5,10,15,20-tetraphenylporphyrin; P5 (5,10,15,20-tetraphenylporphinato) dichlorophosphorus(V) chloride, P6 5,10,15,20-tetrakis-(N-methyl-4-pyridyl) porphyrin-Zn(II) tetrachloride, P7 Zn(II)-5,10,15,20-meso-tetrakis-(4-aminophenyl)porphyrin. The meso-porphyrin derivatives were screened for their antimicrobial activity against six reference strains: Streptococcus pyogenes ATCC 19615, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 700603, Pseudomonas aeruginosa ATCC27853 and Candida albicans ATCC 10231. The antimicrobial activity of these samples was evaluated by the agar disk diffusion method and dilution method, with the determination of the minimum inhibitory concentration (MIC), the minimum bactericidal concentration (MBC) and the minimum fungicidal concentration (MFC). The most significant result is provided by the water-soluble P5 manifesting an obvious antimicrobial activity against Streptococcus pyogenes. On the other hand, P6 is a moderately active derivative against Streptococcus pyogenes and Escherichia coli and P7 presents moderate activity against Streptococcus pyogenes and Staphylococcus aureus. All the tested porphyrin bases, presenting hydrophobic character, have no antimicrobial activity under the investigated conditions. The common characteristics of the porphyrins that act as promising antimicrobial agents in the non-irradiated methods are: the cationic nature, the increased hydrophilicity and the presence of both amino functional groups grafted on the porphyrin ring and the coordination with Zn or phosphorus in the inner core.

The development of factor VIII inhibitors (allo-antibodies) continues to be a major complication in the management of severe forms of hemophilia A, especially as far as treatment and treatment response monitoring is concerned. The need to implement a reliable laboratory assay is all the more obvious if major surgery occurs, when conventional tests (activated partial thromboplastin time APTT, prothrombin time PT, factor VIII level) are of no avail and there is a very fragile balance between bleeding and thrombosis.

We report the case of a 32 year-old patient diagnosed with severe Hemophilia A, referred to the Comprehensive Center for the Diagnosis and Treatment of Hemophilia of the Fundeni Clinical Institute for a multidisciplinary assessment in view of a total left hip arthroplasty due to aseptic necrosis of the femoral neck.

Workup showed a high inhibitor titer (>200 BU). Taking into consideration the interindividual variability of the response to bypassing agents, as well as the bleeding risk associated with a major orthopedic surgery, we used thromboelastography (TEG) to assess the patient’s response to aPCC (activated prothrombin complex concentrate) and rFVIIa (activated recombinant factor VII). The findings helped select the optimal replacement scheme to ensure perioperative hemostasis.