Nutritional therapy for hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the most frequent primary liver cancer and presents together with cirrhosis in most cases. In addition to commonly recognized risk factors for HCC development, such as hepatitis B virus/hepatitis C virus infection, age and alcohol/tobacco consumption, there are nutritional risk factors also related to HCC development including high intake of saturated fats derived from red meat, type of cooking (generation of heterocyclic amines) and contamination of foods with aflatoxins. On the contrary, protective nutritional factors include diets rich in fiber, fruits and vegetables, n-3 polyunsaturated fatty acids and coffee. While the patient is being evaluated for staging and treatment of HCC, special attention should be paid to nutritional support, including proper nutritional assessment and therapy by a multidisciplinary team. It must be considered that these patients usually develop HCC on top of long-lasting cirrhosis, and therefore they could present with severe malnutrition. Cirrhosis-related complications should be properly addressed and considered for nutritional care. In addition to traditional methods, functional testing, phase angle and computed tomography scan derived skeletal muscle index-L3 are among the most useful tools for nutritional assessment. Nutritional therapy should be centered on providing enough energy and protein to manage the increased requirements of both cirrhosis and cancer. Supplementation with branched-chain amino acids is also recommended as it improves response to treatment, nutritional status and survival, and finally physical exercise must be encouraged and adapted to individual needs.     

Genetic Variation of Echinococcus canadensis (G7) in Mexico

We evaluated the genetic variation of Echinococcus G7 strain in larval and adult stages using a fragment of the mitochondrial cox1 gen. Viscera of pigs, bovines, and sheep and fecal samples of dogs were inspected for cystic and canine echinococcosis, respectively; only pigs had hydatid cysts. Bayesian inferences grouped the sequences in an E. canadensis G7 cluster, suggesting that, in Mexico, this strain might be mainly present. Additionally, the population genetic and network analysis showed that E. canadensis in Mexico is very diverse and has probably been introduced several times from different sources. Finally, a scarce genetic differentiation between G6 (camel strain) and G7 (pig strain) populations was identified.Echinococcus granulosus sensu lato (s.l.) includes species that cause cystic echinococcosis (CE), one of the most important and widespread parasitic zoonoses. Recent phylogenetic studies based on both mitochondrial and nuclear DNA genes show that E. granulosus s.l. consists of at least four valid species: E. granulosus sensu stricto (s.s.; genotypes G1–G3), E. equinus (G4), E. ortleppi (G5), and E. canadensis (G6–G10). Genotypes G6/G7 are closely related and referred to as camel and pig strains, respectively.^1–3 The pig–dog cycle is mainly present in Mexico and maintains the G7 strain.^4,5 Although there are isolated reports of E. oligarthrus in a wild cat,⁶ E. ortleppi (E. granulosus s.l.; G5) in a patient,⁷ and E. granulosus s.s. (G1) in a rural pig, there is no evidence that these species are maintained in Mexico.⁸ No data of CE caused by G7 have been documented in Mexican patients, although there is a high number of E. canadensis G7-infected patients in central Europe, pointing to the importance of this strain as a cause of human CE.^9,10 There are only two genetic studies performed in samples from Mexico. Cruz-Reyes and others⁵ documented that G7 parasites of Mexican and Polish pig isolates showed similar patterns by restriction fragment length polymorphism (RFLP) of ribosomal DNA (rDNA) internal transcribed spacer 1 (ITS1) and random amplified polymorphic DNA (RAPD) techniques, and although polymerase chain reaction (PCR) -sequencing analysis of mitochondrial cox1 gen fragment was performed, no polymorphism data were reported. Sharma and others¹¹ identified two variants (A and B) inside of the G6/G7 group consisting of samples from Mexico and Argentina using five nuclear markers (elongation factor 1α, transforming growth factor-β receptor kinase, thioredoxin peroxidase, calreticulin, and ezrin-radixin-moesin-like protein). Because some local slaughter records from northern Mexico indicate the presence of Echinococcus spp. in livestock animals,⁵ the objective of this study was to investigate if parasites in pigs and dogs correspond to G7 and if so, describe its genetic variation.Infected animals were identified in the municipal slaughterhouse of Calera, Zacatecas (north central Mexico), where farm and backyard livestock animals coming from the whole state and other surrounding states were included. For this purpose, viscera from 387 pigs, 243 bovines, and 32 sheep were inspected for the larval stage of Echinococcus. Nine pigs (six pigs from Zacatecas, two pigs from Aguascalientes, and one pig from Morelos) were found infected, and hydatid cysts were obtained under aseptic conditions. After cyst contents were aspirated and centrifuged, aliquots were examined under microscopy to confirm the presence of protoscolices, and pellets were kept in 70% ethanol at −20°C until DNA extraction. Each cyst from each animal was considered as an isolate.Based on the presence of the parasites previously identified in Calera''s slaughterhouse, a rural community located in the central area of Zacatecas at 22°55′ N, 102°48′ W was selected to look for the adult stage of this parasite. For this search, all dogs (60) present in the community were sampled one time for feces after obtaining verbal consent from the owner; samples were used to identify taeniid eggs by the Faust technique, antigens in stool samples (copro-antigens) by enzyme-linked immunosorbent assay (ELISA; CpAg ELISA), and DNA by Copro-PCR. The CpAg ELISA was performed as described by Allan and others¹² and Moro and others.¹³ For Copro-PCR, only positive samples by CpAg ELISA were analyzed using JB3 and JB4 primers to amplify a cox1 gen fragment.¹⁴ Coprological analysis of dogs showed that 11 samples were positive by CpAg ELISA (18.3%); only 2 of these samples had taeniid tapeworms (3.4%), and 3 of 11 samples yielded products of approximately 450 bp. All amplicons obtained of hydatid cysts and fecal samples were purified, sequenced on both strands, submitted to GenBank (accession numbers {"type":"entrez-nucleotide-range","attrs":{"text":"KF734649-KF734660","start_term":"KF734649","end_term":"KF734660","start_term_id":"570963772","end_term_id":"570963794"}}KF734649-KF734660), and compared with several mitochondrial DNA sequences of cox1. Dogs positive for taeniid eggs or antigens were purged and treated with praziquantel at 30 mg/kg and arecoline bromide at 2 mg/kg. The protocol was previously approved by the Ethics and Research Committees of the General Hospital “Dr. Manuel Gea Gonzalez”; government and health authorities of the municipality and community also authorized our study.All sequences were subjected to the Basic Local Alignment Search Tool (BLAST) search in the GenBank database; multiple alignments were performed with the CLUSTAL W and MUSCLE programs,^15,16 with manual adjusted in MEGA program v5¹⁷ to determine the appropriate model of molecular evolution in the Modeltest 3.7 program.¹⁸ The phylogenetic reconstruction using Bayesian inference was performed with Mr Bayes 3.2.1 program.¹⁹ Unrooted haplotype networks were created using NETWORK 4.611 software and nested according to the rules in median-joining networks.²⁰ An analysis of genetic diversity within and between populations was performed using DnaSPv4²¹ and included nucleotide diversity (π), haplotype polymorphism (θ), genetic differentiation index (F_ST), and Tajima''s D test. Analysis of molecular variance (AMOVA) was used to examine the population genetic structure between populations by Φ_ST as the genetic fixation index (analogous to F_ST) obtained by ARLEQUIN software.²²After multiple alignments, all sequences of larval and adult stages showed 98% or higher identity with E. canadensis, whereas the Bayesian phylogenetic tree and the haplotype network inference grouped these sequences in the E. canadensis G7 cluster. Sequences for cox1 of E. canadensis from Africa, Asia, Europe, Latin America, and North America deposited in the GenBank databases (N = 58) as well as our sequences (accession numbers {"type":"entrez-nucleotide-range","attrs":{"text":"KF734649-KF734660","start_term":"KF734649","end_term":"KF734660","start_term_id":"570963772","end_term_id":"570963794"}}KF734649-KF734660) were analyzed. The results for π and θ were 0.0118 and 0.718, and the result of Tajima''s D test was −2.1885 (P < 0.01). Genetic differentiation indexes between different paired sequences of E. canadensis genotypes are shown in

The Adult Macaque Spinal Cord Central Canal Zone Contains Proliferative Cells And Closely Resembles The Human

The persistence of proliferative cells, which could correspond to progenitor populations or potential cells of origin for tumors, has been extensively studied in the adult mammalian forebrain, including human and nonhuman primates. Proliferating cells have been found along the entire ventricular system, including around the central canal, of rodents, but little is known about the primate spinal cord. Here we describe the central canal cellular composition of the Old World primate Macaca fascicularis via scanning and transmission electron microscopy and immunohistochemistry and identify central canal proliferating cells with Ki67 and newly generated cells with bromodeoxyuridine incorporation 3 months after the injection. The central canal is composed of uniciliated, biciliated, and multiciliated ependymal cells, astrocytes, and neurons. Multiciliated ependymal cells show morphological characteristics similar to multiciliated ependymal cells from the lateral ventricles, and uniciliated and biciliated ependymal cells display cilia with large, star‐shaped basal bodies, similar to the Ecc cells described for the rodent central canal. Here we show that ependymal cells with one or two cilia, but not multiciliated ependymal cells, proliferate and give rise to new ependymal cells that presumably remain in the macaque central canal. We found that the infant and adult human spinal cord contains ependymal cell types that resemble those present in the macaque. Interestingly, a wide hypocellular layer formed by bundles of intermediate filaments surrounded the central canal both in the monkey and in the human, being more prominent in the stenosed adult human central canal. J. Comp. Neurol. 522:1800–1817, 2014. © 2013 Wiley Periodicals, Inc.     

Laparoscopic aneurysm resection and splenectomy for splenic artery aneurysm in the third trimester of pregnancy

Background

Splenic artery aneurysms (SAA) are a rare entity most commonly diagnosed incidentally. Their association with pregnancy increases the risk of rupture resulting in a disproportionately high maternal and fetal mortality. Accordingly, elective surgical treatment is recommended in asymptomatic patients with aneurysms less than 2 cm. In this case, we present a patient during her third trimester of pregnancy with a SAA who was treated by laparoscopic aneurysm resection and splenectomy.

Methods

The patient is a 38-year-old multiparous woman, with an incidental diagnosis of a SAA in 2010. Subsequently, the patient became pregnant and at 27 weeks started to develop abdominal pain. Failed embolization was attempted with worsening of the patient’s symptoms. A CT angiogram revealed a 1.6 cm distal third SAA without any evidence of rupture. Due to the localization of the lesion, the patient was offered a laparoscopic aneurysm resection and splenectomy.

Results

Operating time was 90 min and estimated blood loss was 5 cc. Postoperative fetal monitoring was normal. No perioperative complications were observed. The patient was discharged on postoperative day 3. Two months after laparoscopic splenectomy, the patient delivered a male infant in perfect health.

Conclusions

Although this is a rare disease, the risk of aneurysmal rupture is increased during pregnancy. As a result of high maternal and fetal mortality, elective surgery should be performed. Laparoscopic surgery is the technique of choice.     

Arsenic methylation capacity is associated with breast cancer in northern Mexico

Exposure to environmental contaminants, dietary factors and lifestyles may explain worldwide different breast cancer (BC) incidence. Inorganic arsenic (iAs) in the drinking water is a concern in many regions, such as northern Mexico. Studies in several countries have associated the proportion of urinary monomethylarsenic (%MMA) with increased risks for many As-related diseases, including cancer. To investigate the potential relationships between the risk of BC and the capacity to methylate iAs, a hospital-based case–control study (1016 cases/1028 controls) was performed in northern Mexico. Women were directly interviewed about their reproductive histories. The profile of As metabolites in urine was determined by HPLC-ICP-MS and methylation capacity was assessed by metabolite percentages and indexes. Total urinary As, excluding arsenobetaine (TAs-AsB), ranged from 0.26 to 303.29 μg/L. Most women (86%) had TAs-AsB levels below As biological exposure index (35 μg/L). Women with higher %MMA and/or primary methylation index (PMI) had an increased BC risk (%MMA OR_Q5vs.Q1 = 2.63; 95%CI 1.89,3.66; p for trend < 0.001; PMI OR_Q5vs.Q1 = 1.90; 95%CI 1.39,2.59, p for trend < 0.001). In contrast, women with higher proportion of urinary dimethylarsenic (%DMA) and/or secondary methylation index (SMI) had a reduced BC risk (%DMA OR_Q5vs.Q1 = 0.63; 95%CI 0.45,0.87, p for trend 0.006; SMI OR_Q5vsQ1 = 0.42, 95%CI 0.31,0.59, p for trend < 0.001). Neither %iAs nor total methylation index was associated to BC risk. Inter-individual variations in iAs metabolism may play a role in BC carcinogenesis. Women with higher capacity to methylate iAs to MMA and/or a lower capacity to further methylate MMA to DMA were at higher BC risk.     

Body adiposity but not insulin resistance is associated with -675 4G/5G polymorphism in the PAI-1 gene in a sample of Mexican children

ObjectiveTo assess whether the -675 4G/5G polymorphism in the plasminogen activator inhibitor-1 gene is associated with obesity and insulin resistance in Mexican children.MethodsA cross-sectional study was performed in 174 children, 89 with normal-weight and 85 with obesity, aged from 6 to 13 years. All children were from state of Guerrero, and recruited from three primary schools in the city of Chilpancingo, state of Guerrero, Mexico. Insulin levels were determined by immunoenzymatic assay. The homeostasis model assessment was used to determine insulin resistance. The -675 4G/5G polymorphism in PAI-1 gene was analyzed by polymerase chain reaction-restriction fragment length polymorphism.ResultsThe prevalence of insulin resistance in the obese group was higher (49.41%) than in the normal-weight group (16.85%). The 4G/5G PAI-1 polymorphism was found in Hardy Weinberg equilibrium. The 4G/5G genotype contributed to a significant increase in waist-hip ratio (β = 0.02, p = 0.006), waist circumference (β = 4.42, p = 0.009), and subscapular skinfold thickness (β = 1.79, p = 0.04); however, it was not related with insulin resistance.ConclusionThe -675 4G/5G genotype of PAI-1 gene was associated with increase of body adiposity in Mexican children.     

Differential Distribution of Bradykinin B(2) Receptors in the Rat and Human Cardiovascular System

-Bradykinin, a major vasodilator peptide, plays an important role in the local regulation of blood pressure, blood flow, and vascular permeability; however, the cellular distribution of the major bradykinin B(2) receptor in the cardiovascular system is not precisely known. Immunoblot analysis with an anti-peptide antibody to the bradykinin B(2) receptor or chemical cross-linkage with [(125)I]Tyr(0)-bradykinin revealed a band of 69+/-3 kDa at varying intensity in the homogenates of the endothelium and tunica media of the rat aorta and endocardium. Immunostaining showed that the B(2) receptor is abundant in the endothelial linings of the aorta, other elastic arteries, muscular arteries, capillaries, venules, and large veins, where it localizes preferentially to the luminal face of the endothelial cells. In marked contrast, small arterioles (ie, the principal blood-pressure regulating vessels) of the mesenterium, heart, urinary bladder, brain, salivary gland, and kidney had a different staining pattern in which B(2) receptor was prominent in the perivascular smooth muscle cells of the tunica media. A similar distribution pattern was found in mouse as well as in human tissues, indicating that the particular distribution pattern of the B(2) receptor in arterioles is not a species-specific phenomenon. During development, the distribution of B(2) receptor in the heart changes; for example, in the heart of newborn rats, the B(2) receptor was abundant in the myocardium, whereas in the adult heart, the receptor was present in the endocardium of atria, atrioventricular valves, and ventricles but not in the myocardium. Thus, B(2) receptors are localized differentially in different parts of the cardiovascular system: the arterioles have smooth muscle-localized B(2) receptors, and large elastic vessels have endothelium-localized receptors.     

Comparison of 15 different stents in superficial femoral arteries by high resolution MRI ex vivo and in vivo

PURPOSE: To evaluate the MRI compatibility of 15 different commercially available, new generation, U.S. Food and Drug Administration (FDA)-approved stents suitable for deployment in superficial femoral arteries (SFAs), and to identify the ones that permit MRI to visualize the wall and lumen of stented arteries with sufficient spatial and contrast resolution to quantify restenosis after stent placement. MATERIALS AND METHODS: A total of 13 nitinol stents and two stainless-steel stents were placed in excised cadaveric SFAs and imaged by MRI at 1.5 T ex vivo. The images were evaluated qualitatively for the presence of artifacts and for the effects of the stent on image contrast, and quantitatively for the effect on signal-to-noise ratio (SNR) of the lumen of the artery inside the stent compared to the SNR of the fluid outside the artery. A nitinol stent was placed in the SFA of a 60-year-old man and imaged at 1.5 T in vivo. RESULTS: Both the vessel wall and the lumen could be visualized in cadaveric SFAs containing either the Absolute nitinol stent, the Dynalink nitinol stent, or the aSpire nitinol-covered stent. Their inside stent/outside stent SNR was 0.7, 0.8, and 0.8, respectively. The other 10 nitinol stents tested obscured the lumen but did not cause major image shape artifacts. Both stainless-steel stents tested, the WallGraft and WallStent, completely obscured the lumen and caused significant distortion of the image shapes. When the Dynalink stent was inserted into a highly stenosed SFA in vivo, the image showed a dark expanded eccentric lumen, circumscribed by a medium intensity band containing the stent. CONCLUSION: MRI can be used to visualize both the lumen and wall of SFAs containing selected nitinol stents ex vivo and in vivo. These results suggest that MRI can be used to monitor restenosis in stents placed in the femoral arterial bed.