Recombinant NY-ESO-1 cancer antigen: production and purification under cGMP conditions.

The cancer-testis antigen, NY-ESO-1, has been engineered into a bacterial expression plasmid which incorporates a His6-tag. The plasmid was transfected into E. coli strain BL21 and Master and Working cell banks generated from this expression system. Three 15-litre fermentations were performed under cGMP (code of Good Manufacturing Practice) conditions and the crude NY-ESO-1 tagged protein isolated as solubilised inclusion bodies. A three-step cGMP chromatography process (immobilised metal affinity, anion exchange, and hydrophobic interaction) was utilised to purify the protein. The purified NY-ESO-1 is being used in early stage human cancer vaccine trials in Australia and the U.S.A.

Invasive testing for the karyotyping of mid-trimester intrauterine fetal death (IUFD): a pilot study.

INTRODUCTION: Aneuploidy remains a common cause of fetal loss after the first trimester. Conventional karyotyping from fetal solid tissues post-delivery unfortunately has a poor success rate particularly where the fetus is macerated. To overcome this we obtained amniocentesis and/or chorionic villus samples from mid-trimester intrauterine fetal deaths (IUFDs) prior to medical termination of pregnancy. SUBJECTS: Ten women with diagnosed IUFD between 12 and 24 weeks' gestation underwent amniocentesis and/or CVS performed after counselling. RESULTS: Successful karyotypes were obtained in all pregnancies. Five of the ten pregnancies were complicated by aneuploidy (two with trisomy 21, two with trisomy 18, and one with trisomy 13). CONCLUSION: The high rate of aneuploidy (50%) in this small cohort emphasises the need for karyotyping. A successful karyotype in all ten pregnancies demonstrates the value of offering these procedures before a termination of pregnancy. We would recommend the adoption of this approach in the management of IUFD occurring after the first trimester.

Interferon beta1a treatment modulates TH1 expression in gammadelta + T cells from relapsing-remitting multiple sclerosis patients.

A paradigm exists that multiple sclerosis is causally related to dysregulation of TH1 inflammatory cytokines and TH2 antiinflammatory cytokines. The cytokine source(s) that initiate the imbalances are unknown. In this study, gammadelta, CD4, and CD8 T cell receptor-positive (TCR+) cells were isolated from the blood of 26 definitive relapsing-remitting multiple sclerosis patients prior to interferon beta-1a (IFNbeta1a) therapy and following 8-10 weeks of this therapy. The bioactivities of interferon gamma (IFNgamma), interleukin 10 (IL10), and interleukin 12 (IL12) were determined. The concentrations of IFN-gamma, IL10, and IL12 from each cell type did not change significantly with IFNbeta1a treatment. The IL10 secreted by gammadelta TCR+ cells strongly correlated with the IL12 secreted by the same gammadelta TCR+ cells, supporting the paradigm. Furthermore, IFNbeta1a therapy decreased the gammadelta TCR+ cell secretion of TH1 cytokines after 8-10 weeks of therapy.

Post-traumatic first metatarsal reconstruction using coralline hydroxyapatite.

Autogenous bone grafting has been the standard approach to reconstruction of trauma-induced metatarsal defects. However, this treatment has well-known disadvantages related to the harvesting, size, shape, and availability of autografts. The authors used a synthetic hydroxyapatite bone-graft substitute manufactured from a marine coral with a morphology similar to that of cancellous bone in the reconstruction of a large, post-traumatic first metatarsal defect. The authors found grafting with coralline hydroxyapatite to be a safe and effective substitute for autogenous bone grafting in the post-traumatic setting.

A survey of physicians' education in caring for the dying: identified training needs.

BACKGROUND: Physicians receive limited systematic training in caring for dying people. The majority of training focuses on technical skills. METHODS: This study surveyed medical students, residents, fellows, and attending physicians to identify physician needs and current types of training in caring for the terminally ill. The study questionnaire was designed to determine whether and when physicians were trained in caring for dying people, the nature of such training, and possible areas to be included in future training. Also requested was a description of a personal experience involving caring for a terminally ill person. RESULTS: One hundred twenty-three questionnaires were distributed and 90 were returned. Data indicate a lack of standardized training in dealing with terminally ill people and an expressed need for periodic, continuous training. The training needs to be mandatory, be ongoing, and use multiple formats, including the integration of theoretical information and clinical practice. Physicians in this study desired training in the following topic areas: 1) pain management; 2) death and dying; 3) the quality of life and death; 4) DNR status. CONCLUSIONS: Physicians in this study indicated a need for more training in how to care for the terminally ill. A model for training should start in medical school and continue throughout one's career. Providing a framework for lifelong professional development that integrates theoretical information and clinical practice with a multidisciplinary approach to patient care should form the basis of a training model.

Treatment of open fractures.

This article reviews current concepts in treating open fractures along with case studies to illustrate specific treatment procedures. It should be used as a basis for care and provide the fundamentals to successfully treat patients in a trauma setting. The practitioner will find this a challenging and exciting area for professional practice.

Splicing of the VASE exon of neural cell adhesion molecule (NCAM) in human small-cell lung carcinoma (SCLC).

Expression of the neural cell adhesion molecule (NCAM) on small-cell lung carcinoma (SCLC) cell lines and tumour tissue has been investigated. Cell lines were found to express highly sialylated NCAM. Neuraminidase treatment revealed the presence of the 140- and 120-kDa isoforms with differential expression of a 95-kDa protein. Similar data were obtained with SCLC tumour tissues. These results were corroborated by Northern blotting where mRNA of 6.7 and 5.5 kb coding for the 140- and 120-kDa isoforms, respectively, were identified. In a few tumours, a weaker band of 7.4-kb mRNA coding for the 180-kDa NCAM was also identified. This result could not be confirmed biochemically due to shortage of material. Finally, a 5-kb transcript was identified in all SCLC samples examined. The NCAM isoform coded by this mRNA remains unknown. Using the polymerase chain reaction (PCR), we have demonstrated the presence of the VASE mini-exon in some isoforms of SCLC NCAM. The VASE mini-exon sequence in human SCLC differs from the published murine sequence by only one base change. This substitution does not result in altered amino-acid sequence.

Interpersonal cognitive problem-solving among children with mild mental retardation.

In this study we sought to determine whether interpersonal cognitive problem-solving skills could distinguish adjusted from nonadjusted classroom behavior among children with mild mental retardation. Discriminant analyses indicated that, irrespective of age or IQ, adjusted children had a larger number of relevant solutions to common problems as well as a higher ratio of aggressive solutions. Findings suggest that cognitive interpersonal skills may be as important, if not even more important, for this population as for youngsters without mental retardation.

Self-instructional posters: one way to save time and money.

Getting staff to continuing education programs can be difficult due to time, staffing, and cost restraints. A self-instructional module combined with a teaching poster format has had a favorable response from participants. This format has the capability of reaching a higher number of participants and is both time- and cost-effective.

An enzyme immunoassay for isotyping mouse monoclonal antibodies.

A rapid, simply performed and relatively inexpensive enzyme immunoassay for isotyping mouse monoclonal antibodies is described, based on the urease/urea system. Because of the high sensitivity (less than 0.1 microgram/ml of immunoglobulin can be detected in cell culture medium) no treatment of the hybridoma supernatant sample is required prior to assay, and the isotype of a mouse immunoglobulin can be determined in about thirty minutes.

Plantar approach for excision of bilateral soft tissue masses in a child.

A painful soft tissue mass of the plantar aspect of both feet, in a 5-year-old child, is presented. Plantar incisions are ideal in specific cases as long as the surgeon adheres to certain principles, as outlined in this manuscript. A discussion of lipomas is also presented. Optimum exposure is the key to total eradication of soft tissue lesions and to proper maintenance of wound healing. Location of the lesion presented warranted use of the plantar approach. Minimal hypertrophy of the plantar scar was achieved, as well as few symptoms by the patient.

Chronic tenosynovitis of the tibialis posterior tendon and the use of tenography.

Tenosynovitis of the tibialis posterior tendon and the use of the tenogram is presented. The tenogram is used as a diagnostic aid in determining the etiology as well as the course of treatment of a symptomatic tibialis posterior tendon. Various treatments for tenosynovitis have also been described.

Severe infections of the head and neck resulting from gas-forming organisms: report of case.

A case of severe infection of the head and neck associated with gas-forming organisms in a patient with fractures of the mandible is presented. Selected aspects of antibiotic therapy are discussed.

Diabetic osteoarthropathy versus diabetic osteomyelitis.

The differentiation between diabetic osteoarthropathy and osteomyelitis is a very difficult one to make, even in the presence of a pedal ulceration. However, considering present medical costs, the incorrect diagnosis of osteomyelitis can be a costly one, not only for the hospital, but also for the patient. In light of a normal white blood cell count, a benign 67Ga-citrate scan, and two noncontiguous areas of bone destruction, the diagnosis of diabetic osteoarthropathy can be made without the necessity of a bone biopsy.

Chairside electrolytic etching of cast alloys for resin bonding.

A new technique and the instruments for using it for chairside electrolytically etching metal have been described. The technique simplifies the process and permits the dentist to etch castings for resin retention in the operatory. The method also applies to spot etching techniques for fixed restorations and removable partial dentures. Intraoral use of the technique was described.

An evaluation of the use of a pH indicator for the detection of beta-lactamase in enzyme immunoassay.

The use of a mixed pH indicator containing bromocresol purple and bromothymol blue was evaluated for the detection of beta-lactamase activity in enzyme immunoassays (EIA) based on this enzyme. The EIA was found to correlate well with a bioassay for antibodies to tetanus toxoid in sheep sera. Results could be read spectrophotometrically at 450 nm or visually as a colour change from dark blue to yellow. The mixed pH indicator was found to have a number of desirable features including good stability, ease of preparation and a sharp colour change which makes the system suitable for visual determination of titration end points.

Improved high-performance liquid chromatographic separation of urinary paracetamol metabolites using radially compressed columns.

Methods have been adapted for the high-performance liquid chromatographic (HPLC) analysis of urinary paracetamol metabolites on radial compression columns. Enhanced resolution and decreased analysis time were two major advances. Various modifications to existing methods were made to counter the effect of the different C18 surface. Thus in ion suppression HPLC the addition of triethylamine at pH 3.0 (phosphate buffer) was necessary to block residual hydroxyl sites, while in ion-pair HPLC a higher tetrabutyl-ammonium hydroxide concentration of 0.01 M at pH 5.0 was used to enhance selectivity. The methods were successfully applied to the study of the metabolism of paracetamol, its glutathione conjugate and 3-thiomethylparacetamol in Sprague-Dawley rats. 3-Thiomethyl-paracetamol sulphoxide and its glucuronide and sulphate conjugates were shown to be metabolites of both 3-thiomethylparacetamol and paracetamol. 3-Thiomethylparacetamol sulphate was unresolved from the sulphates of paracetamol and 3-methoxyparacetamol in ion-pair HPLC. This raises a previously unrecognised problem in which the peak normally attributed to paracetamol sulphate contains metabolites arising from an oxidative metabolic pathway. Elevated levels of 3-methoxyparacetamol conjugates were found in human overdose urine and to some extent in analgesic nephropathy.