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1.
J Nutr Health Aging ; 12(2): 102-6, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18264636

ABSTRACT

OBJECTIVE: To investigate the validity of a patient kept food diary in relation to weighted intakes and to measure dietary intakes of older people both in hospital and after discharge at home. DESIGN: A randomly selected cohort of hospitalised elderly patients was recruited. All patients were instructed how to keep a record of all food and drink consumed and any leftovers. Food diaries were kept for up to seven days in hospital and for seven days in the community. In 18 consecutive patients dietary records were compared with weighed intake. SETTING: Associate Teaching Hospital, United Kingdom. RESULTS: A total of 116 patients participated in the study (median age 77 years, range 66-86 yrs; 49 female). We found significant correlations between food diary and weighted macronutrient intakes both in hospital and in the community. Overall the food diary predicted within+/-17% weighted energy intakes in 70% of individuals. Compared with the National Diet and Nutrition Survey for free-living elderly people in the UK (1998), we found more or less similar energy and micronutrient intakes in hospital, but lower intakes at home. CONCLUSION: Patients kept food diaries can be used to identify those at risk of undernutrition and monitor those on nutritional support. It can also be used for nutritional education and for achieving dietary goals.


Subject(s)
Diet Records , Energy Intake/physiology , Feeding Behavior , Geriatric Assessment , Nutrition Assessment , Aged , Aged, 80 and over , Cohort Studies , Diet , Diet Surveys , Dietary Carbohydrates/administration & dosage , Dietary Fats/administration & dosage , Female , Hospitalization , Humans , Male , Nutritional Status , Predictive Value of Tests , Risk Assessment , United Kingdom
3.
Protein Eng ; 7(12): 1501-7, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7716162

ABSTRACT

We have used the technique of antibody reshaping to produce a humanized antibody specific for the alpha toxin of Clostridium perfringens. The starting antibody was from a mouse hybridoma from which variable (V) region nucleotide sequences were determined. The complementarity-determining regions (CDRs) from these V regions were then inserted into human heavy and light chain V region genes with human constant region gene fragments subsequently added. The insertion of CDRs alone into human frameworks did not produce a functional reshaped antibody and modifications to the V region framework were required. With minor framework modifications, the affinity of the original murine mAb was restored and even exceeded. Where affinity was increased, an altered binding profile to overlapping peptides was observed. Computer modelling of the reshaped heavy chain V regions suggested that amino acids adjacent to CDRs can either contribute to, or distort, CDR loop conformation and must be adjusted to achieve high binding affinity.


Subject(s)
Antibodies, Monoclonal/chemistry , Bacterial Toxins/chemistry , Bacterial Toxins/immunology , Calcium-Binding Proteins , Immunoglobulin Variable Region/chemistry , Type C Phospholipases , Amino Acid Sequence , Animals , Base Sequence , Binding Sites, Antibody , Clostridium perfringens , Humans , Mice , Models, Molecular , Molecular Sequence Data , Peptides/chemistry , Protein Engineering , Recombinant Fusion Proteins/chemistry , Sequence Alignment
4.
Mol Biotechnol ; 1(3): 251-63, 1994 Jun.
Article in English | MEDLINE | ID: mdl-7859164

ABSTRACT

An expression system is described whereby a gene product is expressed fused to an antibody Fab fragment to form an antibody-like molecule. The antigen binding function of the original antibody is retained and the foreign gene replaces the CH2 and CH3 regions of the heavy chain. The fusion protein is secreted as if it were an antibody, and can be purified using the antigen-binding function of the Fab-like part of the molecule. In principle, any open reading frame can be expressed and it is not necessary to develop an individual purification scheme, or any analytical reagents such as antibodies, for the expressed protein, as both these functions can be performed by the Fab part of the fusion protein. In practice, the nature of the nonantibody part of the fusion influences the efficiency of expression and secretion, and detailed guidance is given on trouble-shooting and maximizing expression.


Subject(s)
Antibodies/genetics , Gene Expression , Amino Acid Sequence , Animals , Antibodies/isolation & purification , Base Sequence , Biotechnology , Blotting, Western/methods , Cell Line , Chromatography, Affinity , Cloning, Molecular/methods , Enzyme-Linked Immunosorbent Assay/methods , Escherichia coli/genetics , Genetic Vectors , Immunoglobulin Fab Fragments/genetics , Mice , Molecular Sequence Data , Plasmids/genetics , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/isolation & purification , Transfection
6.
Methods Mol Biol ; 80: 461-74, 1992.
Article in English | MEDLINE | ID: mdl-22351242

ABSTRACT

Whereas the expression of foreign genes in mammalian cells usually proves successful, the purification of gene products is often a difficult and time-consuming process. The availability of monoclonal antibodies to the foreign protein can greatly assist in small scale purification, but where antibodies are not available, alternatives have to be sought One useful approach involves the fusion of the foreign gene adjacent to a gene segment encoding an antibody heavy chain variable region (1). By transfection of this construct into a cell line producing a compatible light chain or by cotransfection of the fusion product with a light chain gene, an antibody-like molecule can be produced and purified using the corresponding antigen.

7.
Methods Mol Biol ; 10: 461-74, 1992.
Article in English | MEDLINE | ID: mdl-23150339

ABSTRACT

Whereas the expression of foreign genes in mammalian cells usually proves successful, the purification of gene products is often a difficult and time-consuming process. The availability of monoclonal antibodies to the foreign protein can greatly assist in small scale purification, but where antibodies are not available, alternatives have to be sought One useful approach involves the fusion of the foreign gene adjacent to a gene segment encoding an antibody heavy chain variable region (1). By transfection of this construct into a cell line producing a compatible light chain or by cotransfection of the fusion product with a light chain gene, an antibody-like molecule can be produced and purified using the corresponding antigen.

9.
AIDS ; 3(3): 147-54, 1989 Mar.
Article in English | MEDLINE | ID: mdl-2496731

ABSTRACT

This report presents the results of a survey conducted between February and April 1988 to ascertain Ugandans' perception of AIDS as a problem, their knowledge of the disease and how it is spread, the resultant change in their sexual behaviour and their knowledge of, and attitudes towards, condoms. Two hundred and four people (98 women, 106 men) were interviewed from three locations; Kampala (an area with a high incidence of AIDS), Kabale, a town in central Kigezi, and villages in North Kigezi (both relatively low incidence areas). The main points to emerge are: (1) all the respondents were aware of the disease; (2) only 3.6% of respondents mentioned AIDS of their own accord as a problem for Uganda; (3) those in Kampala were most knowledgeable and men in all three areas were generally more informed than women; (4) change in behaviour was more pronounced in those who had known an AIDS victim personally; (5) 100% of men and 79% of women had heard of condoms but only eight men (9.4%) and one woman (1.0%) used condoms regularly for the prevention of AIDS, and (6) there was, nonetheless, a degree of willingness to use condoms once informed that they could reduce the risk of AIDS.


PIP: 204 Ugandans (98 women and 106 men) were surveyed in early 1988 to assess Ugandans' perceptions of acquired immunodeficiency syndrome (AIDS) as a social problem, their knowledge of the diseases and its modes of transmission, resultant changes in their sexual behavior, and their attitudes toward condoms. Respondents were drawn from 3 locations: Kampala, an area with a high incidence of AIDS; Kabale, a town in central Kigezi with a low prevalence of AIDS; and villages in North Kigezi, also relatively low incidence areas. In general, respondents did not consider AIDS to be a major problem in their community or in Uganda as a whole and only those from Kampala perceived AIDS as the most serious disease facing Uganda today. Kampala residents and men in all 3 locations were most informed about AIDS and its transmission. 3-4 correct modes of spread were identified by 31% of respondents in the villages, 32% in Kabale, and 67% in Kampala. 90% knew that the best way to avoid AIDS as to be sexually monogamous. Condoms were suggested as a further preventive measures by 19% of respondents in Kabale, 17% in Kampala, and 1.5% in the villages. 68% of men in Kabale, 52% in the villages, and 14% in Kampala admitted contact with prostitutes. 50% of men and 16% of women in Kampala have been treated for a sexually transmitted disease. In response to the AIDS threat, 32% of those in Kampala, 20% in Kabale, and 31% in the villages claimed to have changed their sexual behavior. 21% of respondents in the villages, 19% in Kabale, and 36% in Kampala would be willing to use condoms to reduce the risk of AIDS, although lack of availability constitutes a major barrier. Since marital infidelity is widespread and socially acceptable in Uganda, an effort should be made to target married men for education on AIDS.


Subject(s)
Acquired Immunodeficiency Syndrome/psychology , Contraceptive Devices, Male , Adolescent , Adult , Attitude to Health , Data Collection , Female , Health Education , Health Knowledge, Attitudes, Practice/statistics & numerical data , Humans , Male , Middle Aged , Sexual Behavior , Uganda
10.
Int J Cancer ; 37(6): 813-7, 1986 Jun 15.
Article in English | MEDLINE | ID: mdl-3519472

ABSTRACT

Well-defined basement membrane laminin was seen in 98/158 (62%) rectal adenocarcinomas stained by an immunoperoxidase method. Only 27 (28%) patients with laminin-positive tumours developed distant metastases, compared with 39 (65%) patients with laminin-negative carcinomas. The corrected 5-year survival rates for patients with laminin-positive and laminin-negative tumours were 65% and 23%, respectively. Twenty-five out of 30 (83%) well-differentiated adenocarcinomas and only 3/15 (17%) poorly differentiated tumours contained basement membrane laminin, with moderately differentiated carcinomas showing intermediate laminin status (70/110, 64%, laminin positive). Forty-three of 60 (72%) of laminin-negative tumours had metastasized to regional lymph nodes. These data suggest that laminin may be a marker for differentiation. However, laminin status yields information about tumour behaviour which is not confined to stage and grade, and multivariate analysis shows that it is a better indicator of prognosis than tumour grade as assessed by conventional histology. Although laminin status alone is a less useful predictor of prognosis than Dukes' stage, a patient with a laminin-positive adenocarcinoma of rectum is 2.7 times as likely to survive 5 years than a patient with a laminin-negative tumour. Assessment of laminin status, together with Dukes' stage is, therefore, commended as a more precise and objective indicator of prognosis than histological degree of differentiation in colorectal carcinoma.


Subject(s)
Adenocarcinoma/metabolism , Basement Membrane/metabolism , Laminin/metabolism , Rectal Neoplasms/metabolism , Adenocarcinoma/pathology , Analysis of Variance , Humans , Immunoenzyme Techniques , Lymphatic Metastasis , Prognosis , Rectal Neoplasms/pathology
11.
Br J Cancer ; 50(1): 51-61, 1984 Jul.
Article in English | MEDLINE | ID: mdl-6378237

ABSTRACT

Using an immunoperoxidase procedure, we have examined the distribution of laminin and fibronectin in normal human large intestinal mucosa and in 50 cases of rectal adenocarcinoma for which extensive clinical follow up was available. In normal tissue, laminin staining was largely restricted to basement membranes, including that underlying the epithelial cells, whereas fibronectin was found in both basement membranes and surrounding connective tissue. In rectal carcinomas, basement membrane-like staining for laminin associated with tumour cells was found in only 27 out of the 50 cases studied. Statistical analysis showed that the presence of laminin-containing basement membranes was correlated with low histological grade (well-differentiated tumours), but not with stage (progression through the bowel wall and the development of lymph node metastases) and, in a highly significant way, with a reduced incidence of distant metastases and increased patient survival. Although fibronectin was found in tumour cell basement membranes where these were present, it was also found in the stroma of all 50 tumours. There was no apparent correlation between the presence of stromal fibronectin and grade, stage or development of metastases. Finally, attention is drawn to some of the technical difficulties in detecting basement membrane antigens in formalin-fixed tissue, the material most frequently available for retrospective study.


Subject(s)
Adenocarcinoma/analysis , Fibronectins/analysis , Laminin/analysis , Rectal Neoplasms/analysis , Adenocarcinoma/pathology , Adenocarcinoma/secondary , Basement Membrane/analysis , Collagen/analysis , Humans , Immunoenzyme Techniques , Intestinal Mucosa/analysis , Neoplasm Staging , Rectal Neoplasms/pathology , Retrospective Studies
12.
Biosci Rep ; 4(3): 223-9, 1984 Mar.
Article in English | MEDLINE | ID: mdl-6722288

ABSTRACT

Type-I procollagen, 14C-biosynthetically labelled, was reduced under denaturing and non-denaturing conditions. Reoxidation to disulphide-linked trimers occurred with non-denatured chains in the presence of an oxidant system containing oxidized and reduced glutathione. Dimeric intermediates were not detected. This reoxidation was accelerated by homogeneous beef liver protein disulphide-isomerase.


Subject(s)
Isomerases/metabolism , Procollagen/metabolism , Animals , Carbon Radioisotopes , Chick Embryo , Dithiothreitol/pharmacology , Glutathione/pharmacology , Kinetics , Macromolecular Substances , Molecular Weight , Procollagen/isolation & purification , Protein Disulfide-Isomerases , Sodium Dodecyl Sulfate/pharmacology , Tendons/metabolism
13.
14.
Anaesthesia ; 37(1): 3-8, 1982 Jan.
Article in English | MEDLINE | ID: mdl-7081646

ABSTRACT

The fibrinolytic effects of six different anaesthetic agents were studied in 36 patients undergoing abdominal hysterectomy. Fibrinolytic activity was measured by dilute whole blood and euglobulin clot lysis times. Concentrations of plasminogen, fibrinogen and fibrin degradation products were also determined. Significant enhancement of fibrinolytic activity, measured as a decrease in clot lysis time, was seen during anaesthesia and surgery with all agents studied. Halothane produced the least effect and the greatest increase in fibrinolytic activity was seen during trichloroethylene anaesthesia. Lumbar extradural anaesthesia also consistently produced enhancement of fibrinolysis. The reasons for these results and their clinical implications are discussed.


Subject(s)
Anesthesia/methods , Anesthetics/pharmacology , Fibrinolysis/drug effects , Adult , Blood Coagulation Tests , Enflurane/pharmacology , Female , Fentanyl/pharmacology , Halothane/pharmacology , Humans , Lidocaine/pharmacology , Middle Aged , Trichloroethylene/pharmacology
15.
Biochem J ; 191(3): 873-6, 1980 Dec 01.
Article in English | MEDLINE | ID: mdl-7283980

ABSTRACT

Protein disulphide-isomerase activity was determined in homogenates of chick-embryo tissues. Activities were highest in tissues active in procollagen synthesis and were maximal at the developmental stage of maximal procollagen synthesis. These variations in protein disulphide-isomerase activity correlate closely with those previously observed for protocollagen hydroxylase activities.


Subject(s)
Isomerases/metabolism , Procollagen/biosynthesis , Animals , Bone and Bones/enzymology , Cartilage/enzymology , Chick Embryo , Disulfides/metabolism , Mixed Function Oxygenases/metabolism , Protein Disulfide-Isomerases , Tendons/enzymology , Tissue Distribution
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