Serum vascular endothelial growth factor as a tumor marker for hepatocellular carcinoma in hepatitis C virus-related cirrhotic patients

BACKGROUNDHepatocellular carcinoma (HCC) accounts for 8.2% of all cancer-related deaths worldwide. Being a vascular tumor, vascular endothelial growth factor (VEGF) plays a vital role in HCC pathogenesis, growth, and spread.AIMTo determine the accuracy of serum VEGF and VEGF/platelet (PLT) as tumor markers in the early detection of HCC cases in patients with hepatitis C virus (HCV)-related liver cirrhosis.METHODSWe conducted a case-control study with HCV patients from the outpatient and inpatient hepatology clinics. Patients were classified into three groups: (1) HCC group; (2) Cirrhosis group; and (3) HCV without cirrhosis (control group). Patients were clinically evaluated, and blood samples were drawn for the analysis; serum VEGF levels were measured by a specific VEGF human recombinant enzyme-linked immunosorbent assay kit. Data from the three study groups were compared by the one-way analysis of variance or Kruskal-Wallis test. Receivers operating characteristic curves were constructed to determine the optimal cut-off values of alpha fetoprotein (AFP), VEGF, and VEGF/PLT that provided the best diagnostic accuracy. The sensitivity and specificity at the optimal cut-off value of each biomarker were then calculated. RESULTSThis study included one hundred patients (HCC, cirrhosis, and control groups: n = 40, 30, 30, respectively). HCC patients had significantly higher serum VEGF and VEGF/PLT levels than the non-HCC groups (P = 0.001). Serum VEGF and VEGF/PLT showed significant positive correlations with and HCC tumor size, stage, vascular invasion, and Child-Pugh classification. Moreover, a VEGF cut-off the value of 250 pg/mL provided 80% sensitivity and 81.7% specificity for discriminating HCC patient from non-HCC patients. Similarly, the ratio of VEGF/PLT provided sensitivity and specificity of 77.5% and 80%, respectively which is higher than the accuracy provided by AFP. The combination of AFP, VEGF, and VEGF/PLT increases the accuracy of diagnosing HCC to > 95%.CONCLUSIONIn HCV patients, serum VEGF and VEGF/PLT separately or in combination with AFP are reliable biomarkers for early and accurate HCC diagnosis.     

Kidney injury molecule-1 (Kim-1): an early biomarker for nephropathy in type II diabetic patients

Renal damage is a serious major microvascular diabetic complication implicated in the death of diabetic patients, which would necessitate the need for new biomarkers to detect early stage of diabetic nephropathy (DN). Kidney injury molecule-1 (Kim-1), a type I transmembrane protein, is undetectable in normal kidneys but markedly induced in proximal tubules after ischemic and toxic injury. So, the present study was conducted to estimate and evaluate Kim-1 as a biomarker for DN. This cross-sectional study was carried out on 60 male and female type II diabetic patients (whose serum creatinine level was less than 2 mg/dL). Diabetic patients were classified as microalbuminuric with nephropathy (urinary albumin was 30–300 mg/dL) and normoalbuminuric without nephropathy (urinary albumin was <30 mg/dL). Twenty matched apparently healthy subjects were included as control group. Patients and controls were assessed for fasting blood glucose, glycosylated hemoglobin (HbA1c), serum creatinine, blood urea nitrogen (BUN), microalbuminuria, and urinary Kim-1. Urinary Kim-1 levels were elevated significantly tenfold in type II diabetic microalbuminuric patients as compared to the control group and normoalbuminuric diabetic patient. Urinary Kim-1 levels were positively correlated with microalbuminuria, serum creatinine, BUN, duration of diabetes, and BMI. Higher urinary Kim-1 level in T2D particularly in those with nephropathy and its correlation with urinary microalbumin, serum creatinine, blood urea, and BUN may reflect the role of Kim-1 as a biomarker for diagnosis and prognosis of diabetic nephropathy among T2D patients taking into account other risk factors.

     

Nucleic acid-mediated intracellular protein delivery by lipid-like nanoparticles

Intracellular protein delivery has potential biotechnological and therapeutic application, but remains technically challenging. In contrast, a plethora of nucleic acid carriers have been developed, with lipid-based nanoparticles (LNPs) among the most clinically advanced reagents for oligonucleotide delivery. Here, we validate the hypothesis that oligonucleotides can serve as packaging materials to facilitate protein entrapment within and intracellular delivery by LNPs. Using two distinct model proteins, horseradish peroxidase and NeutrAvidin, we demonstrate that LNPs can yield efficient intracellular protein delivery in vitro when one or more oligonucleotides have been conjugated to the protein cargo. Moreover, in experiments with NeutrAvidin in vivo, we show that oligonucleotide conjugation significantly enhances LNP-mediated protein uptake within various spleen cell populations, suggesting that this approach may be particularly suitable for improved delivery of protein-based vaccines to antigen-presenting cells.     

Prospective blinded study of somatic mutation detection in cell-free DNA utilizing a targeted 54-gene next generation sequencing panel in metastatic solid tumor patients

Sequencing of the mutant allele fraction of circulating cell-free DNA (cfDNA) derived from tumors is increasingly utilized to detect actionable genomic alterations in cancer.We conducted a prospective blinded study of a comprehensive cfDNA sequencing panel with 54 cancer genes. To evaluate the concordance between cfDNA and tumor DNA (tDNA), sequencing results were compared between cfDNA from plasma and genomic tumor DNA (tDNA). Utilizing next generation digital sequencing technology (DST), we profiled approximately 78,000 bases encoding 512 complete exons in the targeted genes in cfDNA from plasma. Seventy-five patients were prospectively enrolled between February 2013 and March 2014, including 61 metastatic cancer patients and 14 clinical stage II CRC patients with matched plasma and tissue samples. Using the 54-gene panel, we detected at least one somatic mutation in 44 of 61 tDNA (72.1%) and 29 of 44 (65.9%) cfDNA. The overall concordance rate of cfDNA to tDNA was 85.9%, when all detected mutations were considered. We collected serial cfDNAs during cetuximab-based treatment in 2 metastatic KRAS wild-type CRC patients, one with acquired resistance and one with primary resistance. We demonstrate newly emerged KRAS mutation in cfDNA 1.5 months before radiologic progression. Another patient had a newly emerged PIK3CA H1047R mutation on cfDNA analysis at progression during cetuximab/irinotecan chemotherapy with gradual increase in allele frequency from 0.8 to 2.1%. This blinded, prospective study of a cfDNA sequencing showed high concordance to tDNA suggesting that the DST approach may be used as a non-invasive biopsy-free alternative to conventional sequencing using tumor biopsy.     

Effect of molecular weight of amine end-modified poly(β-amino ester)s on gene delivery efficiency and toxicity

Amine end-modified poly(β-amino ester)s (PBAEs) have generated interest as efficient, biodegradable polymeric carriers for plasmid DNA (pDNA). For cationic, non-degradable polymers, such as polyethylenimine (PEI), the polymer molecular weight (MW) and molecular weight distribution (MWD) significantly affect transfection activity and cytotoxicity. The effect of MW on DNA transfection activity for PBAEs has been less well studied. We applied two strategies to obtain amine end-modified PBAEs varying in MW. In one approach, we synthesized four amine end-modified PBAEs with each at 15 different monomer molar ratios, and observed that polymers of intermediate length mediated optimal DNA transfection in HeLa cells. Biophysical characterization of these feed ratio variants suggested that optimal performance was related to higher DNA complexation efficiency and smaller nanoparticle size, but not to nanoparticle charge. In a second approach, we used preparative size exclusion chromatography (SEC) to obtain well-defined, monodisperse polymer fractions. We observed that the transfection activities of size-fractionated PBAEs generally increased with MW, a trend that was weakly associated with an increase in DNA binding efficiency. Furthermore, this approach allowed for the isolation of polymer fractions with greater transfection potency than the starting material. For researchers working with gene delivery polymers synthesized by step-growth polymerization, our data highlight the potentially broad utility of preparative SEC to isolate monodisperse polymers with improved properties. Overall, these results help to elucidate the influence of polymer MWD on nucleic acid delivery and provide insight toward the rational design of next-generation materials for gene therapy.     

Implications of climate change (global warming) for the healthcare system

What is known and Objective: Temperature‐sensitive pathogenic species and their vectors and hosts are emerging in previously colder regions as a consequence of several factors, including global warming. As a result, an increasing number of people will be exposed to pathogens against which they have not previously needed defences. We illustrate this with a specific example of recent emergence of Cryptococcus gattii infections in more temperate climates. Comment: The outbreaks in more temperate climates of the highly virulent––but usually tropically restricted––C. gattii is illustrative of an anticipated growing challenge for the healthcare system. There is a need for preparedness by healthcare professionals in anticipation and for management of such outbreaks, including other infections whose recent increased prevalence in temperate climates can be at least partly associated with global warming. What is new and Conclusion: (Re)emergence of temperature‐sensitive pathogenic species in more temperate climates will present new challenges for healthcare systems. Preparation for outbreaks should precede their occurrence.     

A comparison of wavelet and curvelet for breast cancer diagnosis in digital mammogram

This paper presents a comparative study between wavelet and curvelet transform for breast cancer diagnosis in digital mammogram. Using multiresolution analysis, mammogram images are decomposed into different resolution levels, which are sensitive to different frequency bands. A set of the biggest coefficients from each decomposition level is extracted. Then a supervised classifier system based on Euclidian distance is constructed. The performance of the classifier is evaluated using a 2×5-fold cross validation followed by a statistical analysis. The experimental results suggest that curvelet transform outperforms wavelet transform and the difference is statistically significant.     

The functional evaluation of human peptide/histidine transporter 1 (hPHT1) in transiently transfected COS-7 cells

Recently, the expression of the human peptide/histidine transporter (hPHT1, SLC15A4) mRNA was observed in the GI tract and in Caco-2 cells, suggesting that it may participate in the intestinal absorption of peptide-based agents. This study aims to elucidate the: (i) protein expression pattern of hPHT1 (SLC15A4) in human small intestine; (ii) cloning of the hPHT1 full-length sequence; (iii) functional characterization of hPHT1 in transiently transfected COS-7 cells. The expression of hPHT1 was measured using Western blot and immunohistochemical analysis. The hPHT1 full-sequence was amplified from BeWo cells, inserted into the pcDNA3.1–V5/His TOPO^® plasmid and transiently transfected into COS-7 cells to investigate the uptake kinetics of [³H]histidine and [³H]carnosine. Time, pH and sodium-dependent uptake studies were performed in mock (empty vector) and hPHT1–COS-7 cells. Results demonstrated hPHT1 protein expression in different intestinal regions. Histidine and carnosine uptake was linear in hPHT1–COS-7 cells over 15 min and was found to be pH-dependent. These substrates and valacyclovir showed significantly higher uptake at pH 5.0 in the hPHT1 transients when contrasted to the mock COS-7 cells, whereas glycylsarcosine uptake was significantly lower and unaffected by pH. Other di- and tripeptides also showed affinity for hPHT1. This study presents the initial functional characterization, the protein expression of the hPHT1 transporter and provides insight into a potentially different route for increasing peptide and peptide-based drug transport.