The effects of age, genotype and diet on hippocampal subfield iron dysregulation and Alzheimer's disease biomarkers in an ApoE mouse model.

Current theories regarding accumulation of Alzheimer's disease-related deposits of abnormal intra- and extracellular proteins include reactions to inflammation and mitochondrial dysfunction. In this study, we explored whether age, genotype and inflammation via diet have a greater effect on dysregulatory protein accumulation in any particular subfield of the hippocampus. We stained for ferritin, ferroportin, hyperphosphorylated tau and ß-amyloid proteins in the hippocampal region of Apolipoprotein E2 (ApoE2), ApoE3 or ApoE4 mice fed a control diet or a hypothesized inflammation-inducing methionine diet and euthanized at 3, 6, 9 or 12 months. We analysed stains based on hippocampal subfield and compared the protein accumulation levels within each group. We found significantly decreased ferritin expression in ApoE4 mice in the CA1 and Hi regions and decreased ferroportin expression in ApoE4 mice in the Hi region. There was also a significant effect on hyperphosphorylated tau protein levels based upon a given mouse genotype and diet interaction. Additionally, there were nonsignificant trends in each hippocampal subfield of increasing ferroportin and hyperphosphorylated tau after 6 months of age and decreasing ß-amyloid and ferritin with age. This study identified that there are changes in iron regulatory molecules based on genotype in the Hi and CA1 regions. Our findings also suggest a diet-genotype interaction, which affects levels of specific Alzheimer's disease biomarkers in the hippocampus. Additionally, we identified a trend toward increased ability to clear ß-amyloid and decreased ability to clear hyperphosphorylated tau with age in all subfields, in addition to evidence of increasing iron load with time.

Beta-ketothiolase deficiency in a Malaysian infant.

Methylacetoacetyl-coenzyme A thiolase (MAT) deficiency is an autosomal recessive disease caused by a defect of mitochondrial acetoacetyl-CoA thiolase (T2). There is an error of isoleucine catabolism and ketone body utilization due to mutations in the acetyl-Coenzyme A acetyltransferase 1 (ACAT1) gene. We report a case of a 14 months old Sabahan boy with beta deficiency who presented with severe sepsis and ketoacidosis who subsequently recovered.

Identification and functional characterization of antimicrobial peptide from the marine crab Dromia dehaani.

Many naturally occurring biologically active compounds are derived from marine organisms. Brachyuran crab Dromia dehaani is also known as sponge crab. Hence, this study was undertaken to isolate and characterize the antimicrobial peptide from the marine crab hemolymph. Therefore the objectives of this research work was to high light the functions of biologically active peptide Dromidin. Dromidin was identified and confirmed as a new antimicrobial peptide in the hemolymph of the crab D. dehaani. Antimicrobial peptide was characterized and confirmed by using various analytical techniques. ESI-MS analysis of the active antimicrobial hemolymph fraction resulted in the value 513.0 Da. In addition, current findings strongly suggest that this peptide is functionally important against the pathogens.

Surgical and functional outcomes of cochlear implantation in post-lingual and cross-over patients: First 5-year review of the National Ministry of Health Malaysia cochlear implant programme.

INTRODUCTION: There has been a paradigm shift in the management of acquired sensory neural deafness in the past 30years. This is due to the emergence of implantable hearing devices such as the cochlear implant. The objective of this study is to identify surgical and functional outcomes of post-lingual and cross-over patients implanted with a cochlear implant under the National Ministry of Heath Cochlear Implant (CI) Program between 2009-2013. MATERIALS AND METHODS: We retrospectively reviewed all postlingual and cross-over recipients of cochlear implants under the National Ministry of Heath CI Programme from 2009 to 2013. The outcomes measured were surgical complications and functional outcome. Surgical complications were divided into major and minor complications. Functional outcomes were measured using Categorical Auditory Performances (CAP) scale. RESULTS: A total of 41 post-lingual and 15 cross-over patients were implanted between 2009 and 2013. The age of implantees ranged from 3.6 years to 63.2 years old. There were two major complications (3.6%), one is a case of electrode migration at three months post implantation, and six months post second implantation. Another was a case of device failure at about one-year post implantation. Both patients were reimplanted in the same ear. There was no minor complication. The CAP score for both groups (overall) showed significant improvement with 96.4% achieved CAP score of five and above at 24 months after implantation (p<0.001). The CAP score showed marked improvement at the first 6 months post implantation and continued to improve with time in both groups. CONCLUSION: The Malaysian National Ministry of Health Cochlear implant (CI) Program between 2009-2013 has been a successful programme with good surgical and functional outcomes among the post lingual and cross-over patients.

Reverse-ChIP Techniques for Identifying Locus-Specific Proteomes: A Key Tool in Unlocking the Cancer Regulome.

A phenotypic hallmark of cancer is aberrant transcriptional regulation. Transcriptional regulation is controlled by a complicated array of molecular factors, including the presence of transcription factors, the deposition of histone post-translational modifications, and long-range DNA interactions. Determining the molecular identity and function of these various factors is necessary to understand specific aspects of cancer biology and reveal potential therapeutic targets. Regulation of the genome by specific factors is typically studied using chromatin immunoprecipitation followed by sequencing (ChIP-Seq) that identifies genome-wide binding interactions through the use of factor-specific antibodies. A long-standing goal in many laboratories has been the development of a 'reverse-ChIP' approach to identify unknown binding partners at loci of interest. A variety of strategies have been employed to enable the selective biochemical purification of sequence-defined chromatin regions, including single-copy loci, and the subsequent analytical detection of associated proteins. This review covers mass spectrometry techniques that enable quantitative proteomics before providing a survey of approaches toward the development of strategies for the purification of sequence-specific chromatin as a 'reverse-ChIP' technique. A fully realized reverse-ChIP technique holds great potential for identifying cancer-specific targets and the development of personalized therapeutic regimens.

Bicarbonate boosts flash response amplitude to augment absolute sensitivity and extend dynamic range in murine retinal rods.

Rod photoreceptors in the retina adjust their responsiveness and sensitivity so that they can continue to provide meaningful information over a wide range of light intensities. By stimulating membrane guanylate cyclases in the outer segment to synthesize cGMP at a faster rate in a Ca2+-dependent fashion, bicarbonate increases the circulating "dark" current and accelerates flash response kinetics in amphibian rods. Compared to amphibian rods, mammalian rods are smaller in size, operate at a higher temperature, and express visual cascade proteins with somewhat different biochemical properties. Here, we evaluated the role of bicarbonate in rods of cpfl3 mice. These mice are deficient in their expression of functional cone transducin, Gnat2, making cones very insensitive to light, so the rod response to light could be observed in isolation in electroretinogram recordings. Bicarbonate increased the dark current and absolute sensitivity and quickened flash response recovery in mouse rods to a greater extent than in amphibian rods. In addition, bicarbonate enabled mouse rods to respond over a range that extended to dimmer flashes. Larger flash responses may have resulted in part from a bicarbonate-induced elevation in intracellular pH. However, high pH alone had little effect on flash response recovery kinetics and even suppressed the accelerating effect of bicarbonate, consistent with a direct, modulatory action of bicarbonate on Ca2+- dependent, membrane guanylate cyclase activity.

Comparison of the Efficacy of Pregabalin and Gabapentin for Preemptive Analgesia in Laparoscopic Cholecystectomy Patients: A Randomised Double-Blind Study.

Introduction Preemptive analgesia is now an essential step of perioperative pain management. Pregabalin and gabapentin, which are drugs primarily used in the treatment of neuropathic pain, are now being contemplated for use as preemptive analgesics. This study aimed to assess the effectiveness of gabapentin and pregabalin as preemptive analgesics. The primary objective of the study was to compare pregabalin and gabapentin versus placebo with regard to a visual analogue scale (VAS) score for postoperative pain for 24 hours, time to first rescue analgesia, and total analgesic consumption over 24 hours. The level of sedation with the help of a modified Ramsay sedation score was also compared. Methods This randomised, double-blind study was conducted on 90 patients aged 18-60 years of the American Society of Anesthesiologists (ASA) physical status I and II undergoing elective laparoscopic cholecystectomy under general anaesthesia at a tertiary health care institute. The patients were randomly divided into three groups of 30 each, namely, Group A (gabapentin - oral two capsules of 300 mg gabapentin), Group B (pregabalin - oral two capsules of 150 mg pregabalin), and Group C (placebo - oral two capsules). The various parameters that were recorded in both groups included a VAS score for pain, total dose of tramadol consumed in 24 hours, modified Ramsay sedation scores in the immediate postoperative period, and adverse effects related to the study drugs (at zero and one hour and two, four, six, 12, and 24 hours). The data were analysed using the Statistical Package for the Social Sciences (SPSS) (version 25; IBM SPSS Statistics for Windows, Armonk, NY) software. Results VAS scores were significantly lower in groups A and B when compared to Group C. However, the scores were comparable in Group A (gabapentin) and Group B (pregabalin). The difference in the mean time of rescue analgesia was statistically highly significant when Group A (gabapentin) was compared with Group C (placebo) (P value<0.001) and when Group B (pregabalin) was compared with Group C (placebo) (P value<0.001). Thus, gabapentin and pregabalin provide a longer postoperative pain-free period (382.6 min and 502.3 min, respectively) when compared to the placebo group (137.8 min). Moreover, the mean dose of tramadol consumption in 24 hours was significantly lower in pregabalin (170 mg) and gabapentin groups (176.7 mg) when compared to the placebo group (286.7 mg). However, there was no significant difference in the total tramadol consumption between the gabapentin and pregabalin groups. The level of sedation up to six hours postoperatively was higher in Group B (pregabalin) and Group A (gabapentin) patients compared to Group C (placebo). On comparing the mean Ramsay sedation scores of Group A (gabapentin) versus Group C (placebo) and Group B (pregabalin) versus Group C (placebo), it was found that there was a highly significant difference at zero and one-hour time intervals (P value<0.001 in both comparisons). Conclusion A single preoperative dose of pregabalin 300 mg or gabapentin 600 mg can be used for effective preemptive analgesia in patients undergoing laparoscopic cholecystectomy.

An investigation to determine the normal 1st to 5th metatarsal peak pressure and PTI ratio of the forefoot.

BACKGROUND: The foot support has been described as that of a tripod. Biomechanical studies often report on peak pressures and pressure time integrals under specific areas of the foot. Reference needs to be made to the normal tripod distribution especially in the forefoot. In the forefoot the ratio between the 1st metatarsal and the 5th metatarsal on the medial and lateral columns of the foot respectively provide an excellent reference especially if the normal ratio is known. This study provides conclusive evidence of the 1st to 5th metatarsal ratio for peak pressures and pressure time integrals in the normal foot to be used as a reference. METHODS: A group of normal healthy volunteers (n = 12) and a group of patients with unilateral end stage hallux rigidus (n = 17) were recruited. Repeated measures of 1st and 5th metatarsal peak pressures and Pressure time integrals were measured to determine reliability of measurement and to provide a reference normal ratio. RESULTS: In the healthy volunteer group, the 1st/5th metatarsal ratio for PTI was very close to 1 while it was just over 1.5 for peak pressure. In patients with unilateral end stage hallux rigidus, in the normal foot the 1st/5th metatarsal ratio for PP and PTI was very close to 1. These ratios were reproduced in the operated foot following 1st MTPJ replacement. CONCLUSION: This study using normal healthy volunteers and patients undergoing 1st MTPJ replacement has demonstrated that the normal 1st/5th metatarsal peak pressure and pressure time integrals should be close to 1.

Phenotypic characteristics of peripheral immune cells of Myalgic encephalomyelitis/chronic fatigue syndrome via transmission electron microscopy: A pilot study.

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a complex chronic multi-systemic disease characterized by extreme fatigue that is not improved by rest, and worsens after exertion, whether physical or mental. Previous studies have shown ME/CFS-associated alterations in the immune system and mitochondria. We used transmission electron microscopy (TEM) to investigate the morphology and ultrastructure of unstimulated and stimulated ME/CFS immune cells and their intracellular organelles, including mitochondria. PBMCs from four participants were studied: a pair of identical twins discordant for moderate ME/CFS, as well as two age- and gender- matched unrelated subjects-one with an extremely severe form of ME/CFS and the other healthy. TEM analysis of CD3/CD28-stimulated T cells suggested a significant increase in the levels of apoptotic and necrotic cell death in T cells from ME/CFS patients (over 2-fold). Stimulated Tcells of ME/CFS patients also had higher numbers of swollen mitochondria. We also found a large increase in intracellular giant lipid droplet-like organelles in the stimulated PBMCs from the extremely severe ME/CFS patient potentially indicative of a lipid storage disorder. Lastly, we observed a slight increase in platelet aggregation in stimulated cells, suggestive of a possible role of platelet activity in ME/CFS pathophysiology and disease severity. These results indicate extensive morphological alterations in the cellular and mitochondrial phenotypes of ME/CFS patients' immune cells and suggest new insights into ME/CFS biology.

Clinical utility of ultrasound versus magnetic resonance imaging for deciding to proceed with uterine artery embolization for presumed symptomatic fibroids.

AIM: To compare the diagnostic utility of pelvic ultrasound (US) and magnetic resonance imaging (MRI) on the clinical decision to proceed with uterine artery embolization (UAE). MATERIALS AND METHODS: Over 2 years, 180 consecutive women (mean age 43) sought consultation for UAE, 116 underwent pelvic US and MRI before possible UAE. US was performed prior to MRI. Imaging was analysed for leiomyoma quantity, size and location, uterine volume, and the presence of potential contraindications to UAE. Discrepancies between imaging methods and cases where discrepancies could have altered management, were recorded. RESULTS: For the 116 patients who completed imaging, the average uterine volume was 701 cm(3) using MRI versus 658 cm(3) using US (p=0.48). The average dominant leiomyoma volume was 292 cm(3) using MRI versus 253 cm(3) using US (p=0.16). In 14 (12.1%) patients US did not correctly quantify or localize leiomyomas compared with MRI (p=0.0005). Thirteen patients did not undergo UAE (patient preference n=9, pre-procedural imaging findings n=4). In the four cases where UAE was not performed due to imaging findings, relevant findings were all diagnosed by MRI compared with two by US (p=0.5). The two cases not detected by ultrasound were adenomyosis and a pedunculate subserosal leiomyoma. Of the 103 patients who underwent UAE, 14 were treated (without complication) despite the presence of a relative contraindication; all 14 relative contraindications were identified by MRI compared with 13 by US (p=1.0). CONCLUSION: MRI is more accurate than US for characterizing uterine leiomyomas. In a small but statistically insignificant number of cases, MRI identified findings that were missed by US, which changed management. For patients that are unsuitable to be assessed with MRI, ultrasound alone is sufficient for pre-UAE assessment.

Frozen Section Quality Assurance.

OBJECTIVES: Composite frozen section turnaround time has limited value, precluding assessment of certain processes: slide preparation (technical) and diagnosis (interpretation). We examined whether measuring these elements could identify delays, hypothesizing that longer times were related to (1) inefficient technical processes and (2) case-specific diagnostic challenges. METHODS: Technical and interpretive times were determined for 1,992 specimens submitted for frozen section in 2017; the data were sorted by surgical specialty. Mean and quartile times were determined for each category with all specimens assessed equally, including those with multiple frozen section blocks. RESULTS: Technical times were significantly longer than interpretive times. Specialty grouping facilitated trend identification and enabled assessment of technical and interpretation challenges. We identified technical issues with certain gross specimens involving overdissection and interpretation delays for specific neoplasms and margins. CONCLUSIONS: Measuring technical and interpretative times and subcategorizing by specialty has aided the assessment of frozen section processing in our laboratory, enabling case isolation for process improvement.

Constitutional haploinsufficiency of tumor suppressor genes in mentally retarded patients with microdeletions in 17p13.1.

Chromosome microdeletions or duplications are detected in 10-20% of patients with mental impairment and normal karyotypes. A few cases have been reported of mental impairment with microdeletions comprising tumor suppressor genes. By array-CGH we detected 4 mentally impaired individuals carrying de novo microdeletions sharing an overlapping segment of approximately 180 kb in 17p13.1. This segment encompasses 18 genes, including 3 involved in cancer, namely KCTD11/REN, DLG4/PSD95, and GPS2. Furthermore, in 2 of the patients, the deletions also included TP53, the most frequently inactivated gene in human cancers. The 3 tumor suppressor genes KCTD11, DLG4, and GPS2, in addition to the GABARAP gene, have a known or suspected function in neuronal development and are candidates for causing mental impairment in our patients. Among our 4 patients with deletions in 17p13.1, 3 were part of a Brazilian cohort of 300 mentally retarded individuals, suggesting that this segment may be particularly prone to rearrangements and appears to be an important cause (approximately 1%) of mental retardation. Further, the constitutive deletion of tumor suppressor genes in these patients, particularly TP53, probably confers a significantly increased lifetime risk for cancer and warrants careful oncological surveillance of these patients. Constitutional chromosome deletions containing tumor suppressor genes in patients with mental impairment or congenital abnormalities may represent an important mechanism linking abnormal phenotypes with increased risks of cancer.

Comparison of LVEF obtained with single-plane RAO ventriculography and echocardiography in patients with and without obstructive coronary artery disease.

The left ventricular ejection fraction (LVEF) determined by invasive ventriculography (routine cardiac cath; LV-gram) was compared with that determined by echocardiography in 100 patients scheduled for angiography (86% had LV-gram and 2DE during same hospital admission). Seventy percent of patients had at least single-vessel obstructive coronary artery disease, defined as more than 50% stenosis. By all estimates, the LVEF was higher in patients without coronary artery disease (CAD) compared to patients with CAD. There was an excellent correlation between the LVEF by cath and echo, but this correlation was noticeably less strong in patients with CAD, especially with involvement of the left circumflex artery.

Clinically-relevant postzygotic mosaicism in parents and children with developmental disorders in trio exome sequencing data.

Mosaic genetic variants can have major clinical impact. We systematically analyse trio exome sequence data from 4,293 probands from the DDD Study with severe developmental disorders for pathogenic postzygotic mosaicism (PZM) in the child or a clinically-unaffected parent, and use ultrahigh-depth sequencing to validate candidate mosaic variants. We observe that levels of mosaicism for small genetic variants are usually equivalent in both saliva and blood and ~3% of causative de novo mutations exhibit PZM; this is an important observation, as the sibling recurrence risk is extremely low. We identify parental PZM in 21 trios (0.5% of trios), resulting in a substantially increased sibling recurrence risk in future pregnancies. Together, these forms of mosaicism account for 40 (1%) diagnoses in our cohort. Likely child-PZM mutations occur equally on both parental haplotypes, and the penetrance of detectable mosaic pathogenic variants overall is likely to be less than half that of constitutive variants.

First record of the Gloiopotes huttoni (Thomson, 1890) (Copepoda: Caligidae) parasitic on the swordfish Xiphias gladius along the southeast coast of India.

Parasitic copepods infect a large number of fishes. In particular, the family Caligidae (Crustacea) comprises more than 450 species worldwide, some of which are harmful to cultured and wild fishes. Recently, we found a parasitic copepod on the host swordfish Xiphias gladius (Xiphidae) in the coastal waters of Parangipettai, southeast coast of India. It was identified as Gloiopotes huttoni by its morphological features, and it belongs to the family Caligidae (order Siphonostomatoida). The swordfish was highly infected with about 72 caligids on a single host. Both male and female forms of G. huttoni were colonized on the beak of the swordfish and distributed mostly along the upper and lower jaws of the host, with 42 and 30 copepod parasites, respectively. Most of the copepods were ovigerous, and more females than males were recorded. It is the first record of the occurrence of G. huttoni in India.

Cardiac magnetic resonance imaging investigation of sustained ventricular fibrillation in a swine model--with a focus on the electrical phase.

OBJECTIVES: We sought to develop a method to evaluate the rapidly changing cardiac dimensions during sustained ventricular fibrillation (VF). We also present details of our CPR research imaging program to facilitate this avenue of clinically important research. BACKGROUND: The changes in cardiac dimensions occurring during the initial critical electrical phase of sustained VF are not entirely known. Conventional cardiac magnetic resonance imaging (CMR) functional imaging lacks the temporal resolution necessary to capture the dynamic changes within this early time period of sustained VF. We hypothesized that changes in the middle short axis slice of the ventricles will reflect changes in ventricular volumes accurately. METHODS: Ventricular dimensions were determined from CMR for 30 min of untreated VF in a closed chest, closed pericardium model in seven swine. Ungated steady-state free precession images (SSFP) from the cardiac base to the apex were acquired, taking care to align the anatomical short axis (SAX) imaging planes maximally. The middle slice of the ventricles was determined as the mathematical center of the stack of SAX slices. We then compared the relative changes of right ventricle (RV) and left ventricle (LV) volumes to relative changes in mid-ventricular single slice area. RESULTS: During 30 min of sustained VF, there was an excellent correlation between the changes in exact mid-slice area and the quantitative changes in ventricular volumes (r(2)>0.95). CONCLUSIONS: Mid-slice area data can be used as a surrogate marker of prompt ventricular volume changes during VF. By imaging the heart 10 times faster, the rapid anatomical changes occurring during the initial few minutes of sustained VF can be understood better.

Transarterial chemoembolisation for advanced hepatocellular carcinoma: results from a North American cancer centre.

AIMS: In Asian countries, transarterial chemoembolisation (TACE) has long been used for palliation of unresectable hepatocellular carcinoma (HCC) without strong evidence of improved survival or quality of life. In 2002, a survival benefi of TACE was shown in two randomised controlled trials in Europe and Hong Kong. The effectiveness of interventions fo HCC is influenced by geographical factors related to diverse patient characteristics and protocols. Therefore, the validation of TACE as palliative modality for unresectable HCC requires confirmation in diverse patient populations. The aim of the present study was to assess the effectiveness of TACE for HCC in a North American population. MATERIALS AND METHODS: This was a single centre prospective cohort study. Child-Pugh A cirrhosis or better patients wit unresectable HCC and without radiological evidence of metastatic disease or segmental portal vein thrombosis wer assessed between November 2001 and May 2004. Of 54 patients who satisfied the inclusion criteria, 47 underwent 80 TACE sessions. Chemoembolisation was carried out using selective hepatic artery injection of 75 mg/m(2) doxorubicin and lipiodol followed by an injection of embolic particles when necessary. Repeat treatments were carried out at 2-3 month intervals for recurrent disease. The primary outcome was overall survival; secondary outcomes were morbidity and tumour response. RESULTS: The survival probabilities at 1, 2 and 3 years were 76.6, 55.5 and 50%, respectively. At 6 months after the first intervention, 31% of patients had a partial response and 60% had stable disease by RECIST criteria. Minor adverse events occurred after 39% of TACEs and major adverse events after 20% of sessions, including two treatment-related deaths (4% of patients). One patient had complete cancer remission after undergoing three TACE treatments. Further progression of tumour growth was prevented in 91% of tumours at the 6 month point after the first TACE. At 3 months, serum levels of the tumour marker alpha-feto protein were significantly reduced in patients with elevated levels before TACE. CONCLUSIONS: The survival probabilities at 1 and 2 years after TACE were comparable with results in randomised studies from Europe and Asia. Most patients tolerated TACE well, but clinicians need to be aware that moderately severe sideeffects require close monitoring and prompt intervention.

Cloning and functional characterization of a Na(+)-independent, broad-specific neutral amino acid transporter from mammalian intestine.

We have isolated a cDNA from a rabbit intestinal cDNA library which, when co-expressed with the heavy chain of the human 4F2 antigen (4F2hc) in mammalian cells, induces system L-like amino acid transport activity. This protein, called LAT2, consists of 535 amino acids and is distinct from LAT1 which also interacts with 4F2hc to induce system L-like amino acid transport activity. LAT2 does not interact with rBAT, a protein with a significant structural similarity to 4F2hc. The 4F2hc/LAT2-mediated transport process differs from the 4F2hc/LAT1-mediated transport in substrate specificity, substrate affinity, tissue distribution, interaction with D-amino acids, and pH-dependence. The 4F2hc/LAT2-associated transport process has a broad specificity towards neutral amino acids with K(t) values in the range of 100-1000 microM, does not interact with D-amino acids to any significant extent, and is stimulated by acidic pH. In contrast, the 4F2hc/LAT1-associated transport process has a narrower specificity towards neutral amino acids, but with comparatively higher affinity (K(t) values in the range of 10-20 microM), interacts with some D-amino acids with high affinity, and is not influenced by pH. LAT2 is expressed primarily in the small intestine and kidney, whereas LAT1 exhibits a much broader tissue distribution.