One candidate preparation of human sequence recombinant transforming growth factor-β3 (TGF-β3) was formulated and lyophilized at NIBSC prior to evaluation in a collaborative study for its suitability ...to serve as an international standard. The preparation was tested by 8 laboratories using in vitro bioassays and immunoassays. The candidate preparation 09/234 was judged suitable to serve as an international standard based on the data obtained for biological activity and stability. On the basis of the results reported here, the preparation coded 09/234 was established by the WHO Expert Committee on Biological Standardisation (ECBS) as the WHO 1st IS for human TGF-β3 with an assigned value for TGF-β3 activity of 19, 000IU/ampoule.
One candidate preparation of human sequence recombinant transforming growth factor- beta 3 (TGF- beta 3) was formulated and lyophilized at NIBSC prior to evaluation in a collaborative study for its ...suitability to serve as an international standard. The preparation was tested by 8 laboratories using in vitro bioassays and immunoassays. The candidate preparation 09/234 was judged suitable to serve as an international standard based on the data obtained for biological activity and stability. On the basis of the results reported here, the preparation coded 09/234 was established by the WHO Expert Committee on Biological Standardisation (ECBS) as the WHO 1st IS for human TGF- beta 3 with an assigned value for TGF- beta 3 activity of 19, 000IU/ampoule.
Juvista™ drug product contains human recombinant active transforming growth factor beta 3 (TGFβ3; avotermin). Juvista is being developed for the prevention and reduction of human scarring. Phase II ...and III clinical and development batches of Juvista were assayed for content by an immunoenzymometric assay (IEMA) using a National Institute for Biological Standards and Control (NIBSC) TGFβ3 reference material (98/608) and avotermin standard (Lot 205-0505-005). Paired Juvista TGFβ3 data were compared directly, pooled, and processed using the statistical analysis described by Bland and Altman. A direct comparison of the two standards was also made. The Bland-Altman result was 1.958, the best estimate of the relationship between Lot 205-0505-005 and reference material 98/608. By IEMA, reference material 98/608 has approximately 50% of the immunoreactivity of Lot 205-0505-005. During clinical development, no change in Juvista TGFβ3 dosage was made, but the standard used for Juvista TGFβ3 assay was changed from 98/608 to 205-0505-005. The stated amount of Juvista TGFβ3 in phase III trials was approximately one-half of that in phase II trials. This article highlights the importance of early adoption of an appropriate and representative standard to achieve accurate quantification of protein drug during clinical development.
Celotno besedilo
Dostopno za:
BFBNIB, DOBA, GIS, IJS, IZUM, KILJ, KISLJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
OBJECTIVESUlcerative colitis (UC) is characterized by damage to the intestinal epithelium and connective tissue. The causes of this damage could include changes in the ability of colonic fibroblasts ...to heal wounds and maintain epithelial cell proliferation. Telomeres shorten with each cell division and eventually signal senescence. The aim of this study is to investigate whether the impaired function of rectal fibroblasts in UC is due to accelerated telomere shortening, oxidative stress and premature senescence.
METHODSWe isolated rectal fibroblasts from eight UC patients and nine non-colitis controls, and recorded their in-vitro lifespans. Telomere lengths and superoxide dismutase mRNA expression were also measured by real-time polymerase chain reaction and peroxide levels were measured by flow cytometry.
RESULTSThe fibroblast lifespan decreased as patient age increased (R=0.68, P=0.003) in control patients, but this relationship was absent in UC fibroblasts. We identified a group of patients who were diagnosed later in life than a second group (59 versus 35 years, P=0.002). Fibroblasts from these late-onset UC patients underwent significantly more population doublings before senescence than age-matched controls (25 versus 15, P=0.02). Slower in-vitro telomere shortening rates (32 versus 344, P=0.006) and trends towards longer telomeres at explant were also observed in late-onset UC fibroblasts. Peroxide levels correlated positively with telomere shortening rate (r=0.581, P=0.078).
CONCLUSIONSSome UC-predisposed individuals may have more efficient antioxidant systems that protect the telomeres from oxidative damage. This may allow their rectal fibroblasts to live longer, function better and thus delay the onset of the disease until later life.
Juvista™ drug product contains human recombinant active transforming growth factor beta 3 (TGFβ3; avotermin). Juvista is being developed for the prevention and reduction of human scarring. Phase II ...and III clinical and development batches of Juvista were assayed for content by an immunoenzymometric assay (IEMA) using a National Institute for Biological Standards and Control (NIBSC) TGFβ3 reference material (98/608) and avotermin standard (Lot 205-0505-005). Paired Juvista TGFβ3 data were compared directly, pooled, and processed using the statistical analysis described by Bland and Altman. A direct comparison of the two standards was also made. The Bland-Altman result was 1.958, the best estimate of the relationship between Lot 205-0505-005 and reference material 98/608. By IEMA, reference material 98/608 has approximately 50% of the immunoreactivity of Lot 205-0505-005.
During clinical development, no change in Juvista TGFβ3 dosage was made, but the standard used for Juvista TGFβ3 assay was changed from 98/608 to 205-0505-005. The stated amount of Juvista TGFβ3 in phase III trials was approximately one-half of that in phase II trials. This article highlights the importance of early adoption of an appropriate and representative standard to achieve accurate quantification of protein drug during clinical development.
Celotno besedilo
Dostopno za:
BFBNIB, DOBA, GIS, IJS, IZUM, KILJ, KISLJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Intermittent catheterisation (IC) is a commonly recommended procedure for people with incomplete bladder emptying not satisfactorily managed by other methods. The most frequent complication of IC is ...urinary tract infection (UTI). It is unclear which catheter types, techniques or strategies, affect the incidence of UTI. There is wide variation in practice and important cost implications for using different catheters, techniques or strategies.
To compare sterile versus clean catheterisation technique, coated (pre-lubricated) versus uncoated (separate lubricant) catheters, single (sterile) or multiple use (clean) catheters, self-catheterisation versus catheterisation by others, and any other strategies designed to reduce UTIs in respect of incidence of symptomatic UTI, haematuria, other infections and user preference, in adults and children using intermittent catheterisation for incomplete bladder emptying.
We searched the Cochrane Incontinence Group Specialised Trials Register (searched 19 June 2006), MEDLINE (January 1966 to June 2007), EMBASE (January 1988 to June 2007), CINAHL (January 1982 to June 2007), ERIC (January 1984 to June 2007), the reference lists of relevant articles and conference proceedings, and we attempted to contact other investigators for unpublished data or for clarification.
Randomised controlled trials comparing at least two different catheterisation techniques, strategies or catheter types.
Three reviewers assessed the methodological quality of trials and abstracted data. For dichotomous variables, relative risks and 95% confidence intervals (CI) were derived for each outcome where possible. For continuous variables, mean differences and 95% CI were calculated for each outcome. Because of trial heterogeneity, data were not combined to give an overall estimate of treatment effect.
Fourteen studies met the inclusion criteria; all were small (less than 60 participants). There was considerable variation in length of follow-up and definitions of UTI. Participant drop-out was a problem for several studies. Several studies were more than ten years old and outcome measures varied between studies. Where there were data, confidence intervals around estimates were wide and hence clinically important differences in UTI and other outcomes could neither be identified nor ruled out reliably.
Intermittent catheterisation is a critical aspect of healthcare for individuals with incomplete emptying who are otherwise unable to void adequately to protect bladder and renal health. There is a lack of evidence to state that incidence of UTI is affected by use of sterile or clean technique, coated or uncoated catheters, single (sterile) or multiple use (clean) catheters, self-catheterisation or catheterisation by others, or by any other strategy. The current research evidence is weak and design issues are significant. In light of the current climate of infection control and antibiotic resistance, further, well-designed studies are strongly recommended. Based on the current data, it is not possible to state that one catheter type, technique or strategy is better than another.
The zincins are a superfamily of structurally related zinc-binding metallopeptidases, which play critical roles in biological processes such as sperm-egg fusion, developmental patterning, endocrine ...and immune regulation. These enzymes also have functions in the development and regulation of contractile tissues, which are essential to multicellular organisms for locomotion, cardiovascular function, digestion and reproduction. PRSM-1 is a novel metallopeptidase, isolated in our lab, which is classified as a gluzincin via its HEXXH zinc binding motif and additional upstream glutamic acid. The primary sequence of this protein, however, shares no other homology to the loio wn families of gluzincin metallopeptidases, suggesting that it represents the prototype member of a totally novel family. The recognition of proteins in addition to PRSM-1, by an anti-rPRSM-1 polyclonal serum m western blots, provides initial evidence for the presence of additional PRSM family members. This thesis describes the cloning and characterisation of a novel protein (KG44) which is immunologically related to PRSM-1 and may play a role in the development and differentiation of muscle. Screening of a human placental cDNA expression library with the anti-rPRSM-1 antibodies identified 42 immunoreacting clones, representing 6 distinct cDNAs. Preliminary searches of public sector databases suggested that 3 of these cDNAs were novel and 3 encoded known proteins (including PRSM-1 itself). Antibody preadsorbtion experiments demonstrated that all 3 novel cDNAs, plus one of the previously characterised ones (chaperonin), shared immunological epitopes with PRSM-1. An analysis of the expression of the novel mRNAs in human tissues, revealed that one cDNA (KG14) did dot represent a genuinely expressed gene. The remaining two mRNAs were both widely expressed, but one (KGl 1) was most abundant in human kidney and the other (KG44) was highly upregulated in skeletal muscle and heai't. Neither cDNA was full length and attempts to clone the 5' ends of KGl 1 and KG44 using RACE-PCR were unsuccessful. KG44 was selected for functional characterisation as 70% of the complete cDNA sequence was easily obtainable. Bioinformatic analyses of the deduced amino acid sequence of KG44, did not identify a zincin HDEXXH motif, or any global homology with the zincm metallopeptidases. KG44 is therefore not a novel member of this superfamily and is only related to PRSM-1 via small immunoreactive regions of amino acid sequence. Instead, KG44 is likely to be a novel muscle protem and possible roles for the protein in signal transduction or cell fusion were suggested by the identification of putative phosphorylation sites and a membrane associated amphipathic helix. In order to leai'n more about the function of KG44 in muscle, recombinant protein was expressed in E.coli, purified and used to generate an anti-rKG44 polyclonal antiserum in sheep. In western blots, the KG44 protein was detected in a wide variety of tissues and cell lines, which was consistent with the expression of the mRNA. However, the results were complicated by the appearance of an additional high molecular weight band in tissues and expression of the protein in non-muscle cell lines. Immunohistochemical analysis of fixed human tissue sections detected KG44 in the myofibres of skeletal muscle and in eccrine sweat glands and the walls of small blood vessels in skin.
