Gm Protein Research Articles

Cost-Effective Evaluation of Homemade Recipes in Cancer Patients on Liquid Diet from Low-Middle-Income Group Countries

Cancer patients, on liquid diet are at a high-risk of malnutrition. 10-20% of patients are estimated to die due to malnutrition rather than malignancy itself. In low-middle-income group countries (LMICs), patients are mostly undernourished. They need cost-effective, easily available and acceptable nutritional support. The study has been undertaken to evaluate the cost-effectivity of homemade recipes vs commercially available nutritional formulations for patients on liquid diet. Recipes for both vegetarian and nonvegetarian were designed using low-cost and locally available ingredients to fulfil the daily calorie requirement of ≈2,000 kcal (30 kcal/kg for a 65 kg adult) with ≥90 gm of protein (1.5 gm/kg). Alongside, five commercial nutritional formulations were evaluated to meet the above requirements. The caloric (380 ± 80.1 kcal/100 gm, 260–474 kcal) and protein contents (32.6 ± 10.9/100 gm, 15–49 gm) in commercial supplements were variable. For providing 2,000 kcal with minimum of 90 gm protein, the cost of commercial formulations ranged from INR 745 -3,461/day (1,700 ± 946). Comparatively, the vegetarian and non-vegetarian homemade recipes were estimated to cost just INR 135/day and INR 182/day respectively. For the 2000 kcal diet, the difference in nutrient contents of commercial vs homemade preparations were non-significant, while the cost was significantly lower with homemade recipes (p=0.03). Homemade recipes designed and suited to individual needs could meet the dietary requirements for patients on liquid diet, be highly cost-effective and thereby especially suit patients from resource constraint settings, like LMICs. Keywords: Nutrition, Cancer, Liquid diet, Malnutrition, Low middle­income countries, Nutrition supplement

Nutrition Screening, Reported Dietary Intake, Hospital Foods, and Malnutrition in Critical Care Patients in Malawi.

In low-income countries there are few data on hospital malnutrition. Reduced food intake combined with nutrient-poor foods served in hospitals contribute to nutritional risk. This study investigated whether reported dietary intake and disease state of hospitalized adults in critical care units was related to malnutrition determined by mid-upper arm circumference (MUAC). Adult in-patients (n = 126) in tuberculosis, burn, oncology, and intensive care units in two public tertiary hospitals in Malawi were screened for nutritional status using MUAC and a question on current dietary intake. The hospital menu was reviewed; portion sizes were weighed. The prevalence of moderate and severe malnutrition was 62%. Patients with organ-related diseases and infectious diseases had the highest rates of reduced reported dietary intake, 71.4% and 57.9%, respectively; however, there was no association between reported dietary intake and MUAC. In those unable to eat, however, the rate of severe malnutrition was 50%. The menu consisted of porridge and thickened corn-based starch with fried cabbage; protein foods were provided twice weekly. There was a nutrient gap of 250 calories and 13 gm protein daily. The findings support the need for increasing dietetic/nutrition services to prevent and treat malnutrition in hospitals using simple screening tools.

Theme 6 Tissue biomarkers

Background: Glutamate excitotoxicity is a longstanding hypothesis in the pathophysiology of ALS. Prior studies have demonstrated increased plasma glutamate levels in ALS patients, which suggest a systemic defect in glutamate metabolism (1). The most abundant amino acid consumed in our diet is glutamate. Studies in healthy human subjects have demonstrated efficient metabolism of dietary glutamate via metabolism by enzymes present in the liver, gut lumen and residential gut bacteria. There is increasing evidence that the gut microbiota has significant CNS effects and intestinal dysbiosis has been found in the ALS transgenic SOD1G93A mouse model (2). If intestinal dysbiosis altered the prevalence of glutamine synthetase (GS) producing bacteria, dietary glutamate homeostasis could be impaired, leading to elevated plasma glutamate levels.Objectives: This study examined the degree of fluctuation in plasma glutamate levels seen with consumption of a protein shake in a cohort of ALS patients and family members from a single ALS center.Methods: Twenty six patients (87% of total cohort followed in the ALS center) underwent measurement of plasma amino acid analysis prior to and 1 hour following consumption of a 75 gm protein shake. A subset of 16 patients went on to receive a probiotic with high GS activity and completed serial protein challenges and amino acid analysis during the study. Ten unaffected family members of ALS patients underwent a similar protein challenge. Glutamate Metabolism Dysfunction (GMD) was defined as >30 umol difference post-prandially compared to fasting, and graded as mild (>30-60), moderate (>60-90) or severe (>90).Results: At baseline, 65.4% (17/26) ALS patients screened were GMD positive, compared to 30% (3/10) of tested family members. The severity of GMD in ALS patients was 41% mild, 29% moderate, 29% severe with only mild severity identified in family members. In the six month treatment phase, 75% (6/8) of patients with stable or improving GMD status saw significant improvements in their ALSFRS-R rate of decline, while 71.4% (5/7) with worsening or remaining severe GMD status experienced worsening of their rate of progression.Discussion and conclusions: Although limited by small sample size, this study does represent an excellent sampling within a single ALS center and is the first of its kind to investigate whether impairment in dietary glutamate metabolism exists in ALS patients. If validated in a larger ALS population, detection of glutamate metabolism dysfunction (GMD) could represent a novel biomarker linked to a potential therapeutic target in ALS patients. Planned microbiome analysis will also help in validating this hypothesis.

Revealing Antioxidant and Antidiabetic Potency of Melinjo (Gnetum Gnemon) Seed Protein Hydrolysate at Different Stages of Seed Maturation

roteins hydrolyzed from melinjo seeds (Gnetum gnemon) at green (GM), yellow (YM) and red (RM) stages of maturity were studied for their effectiveness in antioxidant and antidiabetic activities. The seed protein extract was hydrolyzed using alcalase 2.4L, and the resulting hydrolysates with the highest degree of hydrolysis, protein profile, and the most potent contributors to antioxidant and invitro antidiabetic activities were identified. The degree of hydrolysis value of hydrolysates ranged from 52-84%, and the SDS-PAGE protein profile showed two distinct bands in which the band with molecular weight of 30 kDa degraded more intensively. Antioxidant capacity was measured using different standard methods, including radical cation 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate) (ABTS•+) assay, hydroxyl radical (OH•), and superoxide anion (O2•-) scavenging. The green hydrolysate (GMH) had significantly higher (p<0.05) free radical scavenging (ABTS•+, OH•, and O2•-) activities than that of the yellow hydrolysate (YMH) and red hydrolysate (RMH). However, invitro antidiabetic testing was performed based on the inhibitory activity of α-amylase and α-glucosidase. GMH was found to be more effective than YMH and RMH. These results showed that the antioxidant and antidiabetic activity in hydrolyzed GM protein has high potential to be utilized as natural nutraceuticals.

Mass spectrometry combined with affinity probes for the identification of CP4 EPSPS in genetically modified soybeans.

Sample preparation methods used for genetically modified organisms (GMOs) analysis are often time consuming, require extensive manual manipulation, and result in limited amounts of purified protein, which may complicate the detection of low-abundance GM protein. A robust sample pretreatment method prior to mass spectrometry (MS) detection of the transgenic protein (5-enolpyruvylshikimate-3-phosphate synthase [CP4 EPSPS]) present in Roundup Ready soya is investigated. Liquid chromatography-multiple reaction monitoring tandem MS (nano LC-MS/MS-MRM) was used for the detection and quantification of CP4 EPSPS. Gold nanoparticles (AuNPs) and concanavalin A (Con A)-immobilized Sepharose 4B were used as selective probes for the separation of the major storage proteins in soybeans. AuNPs that enable the capture of cysteine-containing proteins were used to reduce the complexity of the crude extract of GM soya. Con A-sepharose was used for the affinity capture of β-conglycinin and other glycoproteins of soya prior to enzymatic digestion. The methods enabled the detection of unique peptides of CP4 EPSPS at a level as low as 0.5% of GM soya in MRM mode. Stable-isotope dimethyl labeling was further applied to the quantification of GM soya. Both probes exhibited high selectivity and efficiency for the affinity capture of storage proteins, leading to the quantitative detection at 0.5% GM soya, which is a level below the current European Union's threshold for food labeling. The square correlation coefficients were greater than 0.99. The approach for sample preparation is very simple without the need for time-consuming protein prefractionation or separation procedures and thus presents a significant improvement over existing methods for the analysis of the GM soya protein.

Development of an Enzyme-Linked Immunosorbent Assay for the Detection of GM Proteins in Transgenic Crops/Produce.

Enzyme-linked immunosorbent assays are always being used extensively for the identification of genetically modified protein in transgenic crops/produce. With the advancement in the detection strategies, competitive and sandwich immunoassays are more convenient and sensitive for the GM protein detection and its quantification in transgenic crops. This chapter is focusing on the competitive ELISA including sandwich ELISA for the detection of the GM proteins.

Effect of Socioeconomic Status on Preeclampsia Cross Sectional Study

AbstractBackground: Preeclampsia (PE) is defined as a pregnancy specific syndrome usually occurs after 20 week gestation, it is determined by increased blood pressure (>140mmHg systolic or >90mmHg diastolic) accompanied by proteinuria (>0.3gm protein in a 24h specimen). Preeclampsia is multifactorial disease with environmental, socioeconomic and seasonal influence. It also has genetic predisposition. Preeclampsia affects approximately 5 to 7% of all pregnancies. And it remains the major cause of maternal and fetal morbidity and mortality.Aim of Study: Is detect the possible association between SES of the pregnant woman and PE and to assess whether the rate of PE or its severity is affected by different SES levels.Patients and Methods: Cross sectional study design on pregnant women diagnosed to have preeclampsia at Luxor International Hospital. The purposive sample of 145 case collected through 6 months during the period from 1st October 2016 until the end of April 2017. Sample size calculated using G power Program version 3.1.3 with Confidence Level (CL) 95%. The study consist of 4 parts. 1st part: A structure inter-viewing questionnaire was done for data collection; personal, medical, obestetrical. 2nd part: Clinical examination to diagnose PE. 3rd part: Laboratory investigation to confirm diagnosis. 4th part: The SES data was designed according to modified Fahmy and El-Sherbini scale 2012 to asses the SES of the participating women which consist of 7 domains.Results: The study resulted in no significant statistical difference between severity of preeclampsia and mother age (p=0.800), obstetric history, previous history of PE (p=0.075) and medical diseases such as essential hypertension (p=0.300) and diabetes mellitus (p=0.316); but there was a significant statistical relationship between severity of PE and bronchial asthma (p=0.008). There is also a significant statistical rela-tionship between severity of PE and level of education (p= 0.024), home sanitation (p=0.014), family possessions (p= 0.0.005), and economic status (p=0.030). but there is no significant statistical relationship between severity of PE and occupation (p=0.111), family domain (p=0.161) and health care domain (p=0.217).Conclusion: There is association between level of educa-tion, home sanitation, family possessions, and economic status as an indicators for socioeconomic status and severity of preeclampsia.Recommendation: Health education programs should be organized for pregnant women about the risk factors of preeclampsia; and for staff members about risk factors, early detection and early management of preeclampsia; improving ante-natal care for pregnant mothers in Egypt and improve the health level of poor families to reduce the risks of preec-lampsia and to help early diagnosis; so that can decrease maternal and perinatal mortality.

Analysis of Select Herpes Simplex Virus 1 (HSV-1) Proteins for Restriction of Human Immunodeficiency Virus Type 1 (HIV-1): HSV-1 gM Protein Potently Restricts HIV-1 by Preventing Intracellular Transport and Processing of Env gp160.

Virus-encoded proteins that impair or shut down specific host cell functions during replication can be used as probes to identify potential proteins/pathways used in the replication of viruses from other families. We screened nine proteins from herpes simplex virus 1 (HSV-1) for the ability to enhance or restrict human immunodeficiency virus type 1 (HIV-1) replication. We show that several HSV-1 proteins (glycoprotein M [gM], US3, and UL24) potently restricted the replication of HIV-1. Unlike UL24 and US3, which reduced viral protein synthesis, we observed that gM restriction of HIV-1 occurred through interference with the processing and transport of gp160, resulting in a significantly reduced level of mature gp120/gp41 released from cells. Finally, we show that an HSV-1 gM mutant lacking the majority of the C-terminal domain (HA-gM[Δ345-473]) restricted neither gp160 processing nor the release of infectious virus. These studies identify proteins from heterologous viruses that can restrict viruses through novel pathways.IMPORTANCE HIV-1 infection of humans results in AIDS, characterized by the loss of CD4+ T cells and increased susceptibility to opportunistic infections. Both HIV-1 and HSV-1 can infect astrocytes and microglia of the central nervous system (CNS). Thus, the identification of HSV-1 proteins that directly restrict HIV-1 or interfere with pathways required for HIV-1 replication could lead to novel antiretroviral strategies. The results of this study show that select viral proteins from HSV-1 can potently restrict HIV-1. Further, our results indicate that the gM protein of HSV-1 restricts HIV-1 through a novel pathway by interfering with the processing of gp160 and its incorporation into virus maturing from the cell.

Distribution of Intact Peanut Protein and Digestion-Resistant Ara h 2 Peptide in Human Serum and Saliva

RATIONALE: Ara h 2 is comparatively resistant to digestion and a digestion-resistant peptide (DRP-Ara h 2) is released in model digestion experiments. The presence of intact peanut protein and/or DRP-Ara h 2 in serum and saliva of humans after ingestion of peanut might help in assessing the risk of an allergic reaction to saliva from kissing or use of shared utensils. METHODS: Ten healthy, non-allergic volunteers (ages 19-50) consumed capsules containing a total of 25 gm of roasted peanut flour (12.75 gm protein). Two additional volunteers, serving as negative controls, consumed capsules containing 25 gm of powdered sugar. Blood and saliva samples were collected at baseline and at various time points up to 24 hours after ingestion. Levels of intact peanut proteins were measured over time using a commercial peanut ELISA. Levels of DRP-Ara h 2 in similar fluids were measured using a uniquely developed inhibition ELISA. RESULTS: DRP-Ara h 2 was detected in the serum of 9 of 10 volunteers (maximum of 248 ng DRP-Ara h 2/ml serum) and in the saliva of 7 of 10 volunteers (maximum of 4890 ng DRP-Ara h 2/ml saliva) for up to 24 hours after ingestion. Intact peanut protein was not detected in either serum or saliva throughout all collection time points. CONCLUSIONS: Demonstration of the stability and distribution of digestion resistant peanut allergens in human body fluids over an extended period of time may provide insight into potential risks posed by exposure to allergenic proteins in saliva.

Well-Characterized Monoclonal Antibodies against Cell Wall Antigen of Aspergillus Species Improve Immunoassay Specificity and Sensitivity

The diagnosis of invasive aspergillosis (IA) based on the detection of Aspergillus galactomannan (GM) is complicated by the presence of cross-reactive GM epitopes in patient specimens. We have developed a novel and specific Aspergillus antigen-capture enzyme-linked immunosorbent assay (ELISA) by the selection of two well-characterized monoclonal antibodies from 17 candidate antibodies. The epitopes recognized by the monoclonal antibodies were present on the cell walls of the hyphae and the conidia of Aspergillus species, which were circulating or excreted as immunodominant antigens during the acute phase of IA established in the animal models. The detection of experimental Aspergillus-mediated antigenemia was suitably sensitive, and the sensitivity was comparable to that of a commercial GM detection ELISA kit (the Platelia Aspergillus assay). Moreover, the specificity of this assay was 100% when it was used to test 382 serum specimens and 120 urine specimens from healthy individuals. Cross-reactivity with other common opportunistic fungi, such as Penicillium and Candida species, and with purified GM protein derived from Aspergillus was not evident. Therefore, the chemical nature of the epitopes captured in this assay is most likely not associated with the GM structure, indicating that this newly developed Aspergillus antigen-capture ELISA is a promising tool for the diagnosis of IA without the risk of the false-positive results that are problematic with current GM antigen assays.

Mating System and Genetic Variations of Tricholoma crassum (Berk.) Sacc. in Some Area of Thailand by Isozyme Electrophoresis and PCR-RFLP Method

Tricholoma crassum (Tricholomataceae, Agaricales) is a large, good edible species. It has nutritional quality, having 10.02 gm carbohydrate, 0.287 gm fats, 18.58 gm protein, 2.71 mg calcium, and 3.35 mg iron in every 100 gm of fresh weight. It is found growing abundantly throughout Thailand.The mating system of T. crassum was studied. The monokaryon isolates were selected from ten sporocarp collections in five provinces in Thailand—Ubonratchathani, Sakonnakorn, Mahasarakham, Srisaket, and Nakornratcha-sima. Suitable growth conditions for the mycelial cultivation were PDYA medium pH 7 and 25 °C incubation. Twelve fast-growing monokaryotic strains were selected for mating system studies. The mating system was determined by pairing the monokaryotic mycelia of each collection in all pairwise combinations. The presence of clamp connections after mating indicated sexual compatibility. The ratio of compatible crosses to all combination crosses were 1:4, which indicated tetrapolar heterothallism. In addition, multiple alleles among the monokaryons for each sporocarp of the five provinces were examined on the basis of their mating interactions, and the results were two factors (A and B), and each possessed 16 different alleles.One hundred and thirty-eight monokarytic isolates of T. crassum from the above five provinces were cultured on growth media. The suitable conditions for the mycelial growth were PDYB medium at 25 °C for 21 days. All samples were analyzed for isozyme variability on polyacrylamide gel electrophoresis with 11 enzyme systems: isocitrate dehydrogenase, leucine aminopeptidase, acid phosphatase, phosphogluconate dehydrogenase, alkaline phosphatase, alcohol dehydrogenase, glucose− 6−phosphate dehydrogenase, lactate dehydrogenase, malate dehydrogenase, laccase, and esterase. Eight of the systems showed polymorphism. Cluster analysis based on isozyme variability using the NTSYSpc 2.00 and UPGMA methods revealed two clusters at a similarity coefficient of 0.67. The first cluster consisted of monokaryotic isolates from Nakornratchasima, Mahasalakham, and eight samples of Ubonratchathani. The second cluster consisted of the isolates from Sakonnakhorn, Srisaket, and 30 samples of Ubonratchathani.The genetic variations of nine additional samples of T. crassum from four additional provinces—Roiyed, Burerum, Patumthani, and Nakhon Pathom—were studied by the technique of PCR-RFLP. Two pairs of primers, ITS1- ITS4 and O1-LR12R, were used respectively for PCR amplification on ITS (internal transcribed spacer) and IGS (intergenic spacer) regions of the nuclear ribosomal gene, followed by digestion with HindIII, DdeI, HaeIII, EcoRI, and HinfI. Data analyses of the PCR-RFLP products based on the similarity index and the UPGMA method in the WinBoot program revealed three clusters that were related to their geographic origins, except the samples from Burirum, which showed genetic variation from the same areas at a similarity coefficient of 0.8 and were grouped into the third cluster.

Decrease in Maximal Force Generation in the Neonatal Mouse Bladder Corresponds to Shift in Myosin Heavy Chain Isoform Composition

A change in calcium handling has been proposed as the cause of decreased maximal force generation by neonatal bladders with growth. Recent studies suggest that increased myosin heavy chain isoform SM1 increases force generation. We studied force generation in neonatal mouse bladders to determine if decreases in SM1 corresponded with decreased force. C57Bl/6 mice were studied from birth to 12 weeks of life (adulthood). The bladder strip contractile response to KCl and bethanechol was followed by the inhibition of rho-kinase activity by Y-27632. The mRNA levels for SM1/SM2 were determined using reverse transcriptase-polymerase chain reaction and protein levels were determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Muscle fraction per cross-sectional area was determined by trichrome staining. Newborn bladders generated significantly more tension in response to KCl (43.3 vs 17.4 mN/mm2, p = 0.02) and bethanechol (40.6 vs 11.9 mN/mm2, p = 0.05) than adult bladders. Inhibition of rho-kinase resulted in similar decreases in tension in all bladders. SM1 mRNA decreased slightly from 60% at birth to 50% at 12 weeks. SM1 protein decreased from 72.5% at birth to 50% by 3 weeks and it remained stable at 12 weeks. Total myosin per gm protein remained stable. Muscle fraction decreased from 63.8% at birth to 58.6% at 12 weeks (p = 0.4). We noted a decrease in SM1 that corresponded to a decrease in bladder force generation. The concept that SM1 contributes to the optimal assembly of myosin filaments suggests that changes in myosin isoforms may have a role in the decrease in voiding pressures seen in normal children.

NEW EMBO MEMBERS' REVIEW: Hsp70 interactions with the p53 tumour suppressor protein

The heat shock proteins (HSPs) are encoded by genes whose expression is substantially increased during stress conditions, such as heat shock, alcohol, inhibitors of energy metabolism, heavy metals, oxidative stress, fever or inflammation. During these conditions, HSPs increase cell survival by protecting and disaggregating stress‐labile proteins (Skowyra et al ., 1990), as well as the proteolysis of the damaged proteins (Wickner et al ., 1999). Under non‐stress conditions, HSPs have multiple housekeeping functions, such as folding and translocating newly synthesized proteins, activation of specific regulatory proteins, including transcription factors, replication proteins and kinases, protein degradation, protein signalling, including steroid hormone activation and tumour immunogenicity, and antigen presentation (for reviews see Helmbrecht et al ., 2000; Jolly and Morimoto, 2000). This broad spectrum of functions gave rise to the term ‘molecular chaperone’, an entity that acts to assist other proteins folding and maturating in the cell. It should also be emphasized that not all HSPs are molecular chaperones and not all chaperones are HSPs (Ellis and Hartl, 1999). HSPs are designated nomenclature according to their approximate molecular weight, e.g. the 70 kDa HSP is known as the molecular chaperone Hsp70. The 70 kDa heat shock‐related proteins comprise a family of highly conserved molecular chaperones that regulate a wide variety of cellular processes during normal and stress conditions (Boorstein et al ., 1994). Hsp70 is one of the most abundant of these proteins, accounting for as much as 1–2% of total cellular protein (Herendeen et al ., 1979). In humans, there are at least 11 distinct genes that code for Hsp70 isoforms, which are located on several different chromosomes (Tavaria et al ., 1996). The major, constitutively expressed hsp70 isoform is called hsc70 (gene product known as the clathrin‐uncoating ATPase or Hsp73) (Welch, 1992). The transcription of inducible forms of hsp70 or hsp72 are under …

Marine Corrinoid-Binding Proteins for the Direct Determination of Vitamin B12 by Radioassay

Under conditions of vitamin B12 deficiency the marine phytoplankton Thalassiosira pseudonana secretes into the growth medium a protein (Gm protein) that binds B12 specifically with an affinity constant of 2 × 1011 M−1. When Gm protein was used as a B12-specific binder in radioassays for the direct determination of the vitamin, the detection limit of the technique approached 5 pg B12/mL. In its natural state, Gm protein is oligomeric having molecular weight of over 400 kDa. Modification of the solution environment of this protein produces unique changes in its tertiary and quaternary structure. The amino acid and submit composition of the protein is reported.

Identification and structure of the Marek's disease virus serotype 2 glycoprotein M gene: comparison with glycoprotein M genes of Herpesviridae family.

We determined the nucleotide sequence of a portion of BamHI-C fragment of Marek's disease virus serotype 2 (MDV2) strain HPRS24 which was suspected to contain the homologue of the herpes simplex virus type 1 (HSV-1) gene UL10, encoding glycoprotein M (gM). An open reading frame whose translation product exhibited significant similarities to HSV-1 gM protein and respective proteins of other herpesviruses of 37.5% and 45.5% to 31.8%, respectively, was identified. A number of distinct transcriptional consensus sequences were found upstream of the first putative start codon of MDV2 UL10 protein. In transcriptional analysis, the gene was transcribed into an 1.5 kb RNA. The primary translation product comprises 424 amino acids with a predicted molecular weight of 46.9 kDa. The predicted MDV2 UL10 protein contains eight hydrophobic domains with sufficient length and hydrophobicity to span the lipid bilayer conserved in the genomes of all herpesviruses which have been sequenced so far. In the region located between the first and second hydrophobic domains, two potential N-linked glycosylation sites were presented. Interestingly, highly charged residues were abundantly possessed in the carboxy-terminal part of the MDV2 UL10 protein. By comparison of the amino acid sequence of the MDV2 UL10 gene with the homologues from other herpesviruses, the data might contribute for further evidence of the evolution of herpesviruses from a common progenitor and an ancient example of MDV2 belonging to the Alphaherpesvirinae subfamily. In addition, the existence of corresponding genes in human, mammalian, and avian herpesvirus genomes, suggests indirectly an important role for gM in the natural life cycle of the virus.

The equine herpesvirus 1 glycoprotein gp21/22a, the herpes simplex virus type 1 gM homolog, is involved in virus penetration and cell-to-cell spread of virions

Experiments to analyze the function of the equine herpesvirus 1 (EHV-1) glycoprotein gM homolog were conducted. To this end, an Rk13 cell line (TCgM) that stably expressed EHV-1 gM was constructed. Proteins with apparent M(r)s of 46,000 to 48,000 and 50,000 to 55,000 were detected in TCgM cells with specific anti-gM antibodies, and the gM protein pattern was indistinguishable from that in cells infected with EHV-1 strain RacL11. A viral mutant (L11deltagM) bearing an Escherichia coli lacZ gene inserted into the EHV-1 strain RacL11 gM gene (open reading frame 52) was purified, and cells infected with L11deltagM did not contain detectable gM. L11deltagM exhibited approximately 100-fold lower titers and a more than 2-fold reduction in plaque size relative to wild-type EHV-1 when grown and titrated on noncomplementing cells. Viral titers were reduced only 10-fold when L11deltagM was grown on the complementing cell line TCgM and titrated on noncomplementing cells. L11deltagM also exhibited slower penetration kinetics compared with those of the parental EHV-1 RacL11. It is concluded that EHV-1 gM plays important roles in the penetration of virus into the target cell and in spread of EHV-1 from cell to cell.

Identification of a 35 kDa glycoprotein from Entamoeba histolytica by sera from patients with amoebic liver abscess and with mouse monoclonal antibody.

In this work, we explored the relevance of a 35 kDa glycoprotein (Gm) of the outer membrane from E. histolytica in the diagnosis of the amoebic liver abscess (ALA) through ELISA and immunoblotting. We were interested in defining the relevance of this antigen in the immune response in patients with amoebic liver abscess and in exploring whether the mouse monoclonal antibody against this 35 kDa glycoprotein recognises the same epitope. We found that 87% of ALA patients had raised antibody levels to Gm antigen, whereas none of the healthy control subjects presented this same increase. We also found 90% sensitivity, 100% specificity, 100% positive predictive value, 90% negative predictive value, and 90% prevalence value for this Gm antigen. Nonetheless, we did not find any statistically significant differences in the levels of immunoglobulins against Gm, although IgG showed a tendency to increase, probably because we are dealing with a secondary immune response. Using electroimmunotransfer blot assay, we found that sera from ALA patients recognise the 35 kDa Gm protein in the same way as it is recognised by the mouse monoclonal antibody, suggesting that is a relevant molecule for the diagnosis of amebiasis, and eventually could lead to its use as protection against the disease.

Cellular bioenergetics, work performance, body composition and psychological indices among women in a diet-exercise program

Forty moderately obese women enrolled in a 10 week study employing a low-fat, rotating energy diet, nutritional supplementation and aerobic walking. Energy intakes rotated between 2.09 and 6.27 MJ (500 and 1500 kcal) on alternate days. On 2.09 MJ days, women consumed meal substitute tablets each hour for the first eight hours of the day (totaling 24 gm carbohydrate, 4 gm fat, 3.2 gm protein), followed by whole food snacks and dinner. On 6.27 MJ days, subjects consumed diets of less than 20 percent fat and engaged in 20 to 30 minutes of aerobic walking. Nutrition supplements were taken daily, 34 women completed the study. Urinary ketones did not change throughout the study. Anthropometric and psychological parameters were measured at weeks 1,3 and 10. Body composition, assessed by bioelectrical impedance, showed mean declines in body weight of 5.8 kg (p<.001), body fat of 3.6 kg (p<.001), percent body fat of 2.4 percent (p<.001), and fat-free mass of 2.1 kg (p<.001). Improvements in reported sense of well-being (p<.01), perceived energy (p<.001) and body image (p<.001) were noted. Skeletal muscle energy function and work output was assessed using a computer-integrated arm ergometer and magnetic resonance spectroscopy to quantify the concentrations of inorganic phosphate (Pi), phosphocreatine (PCr) and intracellular pH (pHi) under resting and exercising conditions. Pre- to post-study increases in maximum arm strength (16.8 to 22.2 kg; pš.001) and total work output (191 to 282 joules; p<.001) were found. No changes over the study period were found in Pi/PCr ratios or pHi under either rest or exercise conditions. Although the impact of specific program components were not identified, the data suggest improved metabolic function in conjunction with favorable changes in psychological indices and body composition.

Higher Calorie Content Preserves Myocardial Electrical Activity During Very‐Low‐Calorie Dieting

We investigated the effect of calorie and protein content of very-low-calorie diets for weight loss upon myocardial electrical activity. Patients were followed on very low calorie diets for up to 20 weeks. Study No. 1 had 16 obese patients (4 men and 12 women). Study No. 2 had 113 obese women. Both study groups were consecutive samples with patients who had BMI > 30. In Study No. 1 patients were given 400 kcal/d with 50 gm of protein and 45 gm of carbohydrates over 12 weeks. In Study No. 2 patients were given either Metabolic Nutrition Program (MNP) which contained 600 kcal/d with 70 gm protein and 70 gm carbohydrates, or Optifast-70 which contained 420 kcal/d with 70 gm protein and 32 gm carbohydrates, for up to 20 weeks. Electrocardiograms were obtained at baseline and during the third month in Study No. 1, and monthly in Study No. 2. The sum of the voltage in leads I+II+III and the length of the QT interval were derived from the electrocardiograms. In Study No. 1 the QRS voltage decreased from 2.77 +/- .18 mv to 2.45 +/- .17 mv (p < .02). Patients lost 18.1 kg +/- 4 kg and the QT interval remained unchanged. In Study No. 2 at 20 weeks the QRS voltage decreased on Optifast-70 (p < .001) in comparison to MNP, in which the QRS voltage remained unchanged. Patients on Optifast-70 lost 17.5 +/- .53 kg, patients on MNP lost 18.5 +/- .66 kg and the QT interval in both groups remained unchanged. The formula diets tested of less than 600 kcal/d were associated with a decrease in electrocardiographic voltage.

Gallbladder emptying stimuli in obese and normal-weight subjects.

Gallbladder stasis may be an important factor in the pathogenesis of cholesterol-gallstone formation in some individuals. We investigated gallbladder function in a group of nondieting, gallstone-free, healthy subjects with normal (22 +/- 1 kg/m2) and high (36 +/- 1 kg/m2) body mass indexes. Fasting gallbladder volume (28.2 +/- 4.4 ml) and residual volume after maximal emptying (8.4 +/- 2.3 ml) in high-body-mass index subjects were not significantly different from those of normal-body-mass index subjects (20.5 +/- 2.5 ml and 4.2 +/- 1.3 ml, respectively). The percentage of gallbladder emptying (71% +/- 5%) and the rate of gallbladder emptying (-1.9 +/- 0.3 x 10(-2) min-1) in high-body-mass index subjects in response to a maximal emptying stimulus was similar to the percentage of emptying (78% +/- 6%) and rate of emptying (-2.3 +/- 0.6 x 10(-2) min-1) in normal-body-mass index subjects. A liquid meal containing less than 1 gm fat, 14 gm protein and 6 gm carbohydrate resulted in both a decreased rate of gallbladder emptying and an increased residual gallbladder emptying and an increased residual gallbladder volume in both groups. The addition of 10 or 20 gm (but not 4 gm) of fat to the liquid meal restored gallbladder emptying to the maximal-stimulus level. These results demonstrate that gallbladder emptying in response to a single liquid meal stimulus is not altered in obesity and that dose-response relationships to fat are similar in obese and normal-weight individuals.(ABSTRACT TRUNCATED AT 250 WORDS)