Comparative study on the deamidation of three recombinant human insulins using capillary electrophoresis.

The applicability of capillary zone electrophoresis (CZE) for the separation of different recombinant human insulins and their deamidated isoforms was studied. The high resolving power of CZE is demonstrated by its ability to separate insulin isoforms differing only by 0.984 Da (different-fold deamidated forms) and even components having the exacts same mass but slightly different shapes (same-fold deamidated forms). From among the several insulins available, humulin, glargine and glulisine were selected for our study because their sequences and chemical parameters are quite similar, however, the small differences present in their amino acid sequences influence the deamidation processes. Using a background electrolyte with basic pH was favourable not only for the separation of the different types of insulin but also for the separation of deamidated protein forms even in a bare fused silica capillary. The LOD values ranged between 0.6 - 0.93 mg/L and 2.17 - 4.37 mg/L for UV and ESI-MS detection, respectively. At -20 - -80 °C, the deamidation is minimal, but at temperatures above +5 °C deamidation is accelerated. At +5 °C only 1-fold deamidation forms could be observed for each insulin. Acidified samples incubated for 1-month at room temperature showed varying levels of deamidation: 1-fold, 1-2-fold and 1-2-3-fold forms for glargine, glulisine and humulin, respectively.

Extracapsular extension in oral cavity cancers-predictive factors and impact on recurrence pattern and survival.

Extracapsular extension (ECE) has long been considered a poor prognostic factor in oral cavity cancer, the presence of which warrants intensification of adjuvant therapy. This study was done to analyze the survival of patients with ECE who received adjuvant chemoradiation. Patients with pathologically confirmed squamous cell carcinoma of the oral cavity, with a minimum of 2 years of follow-up, who were treated at a tertiary cancer centre in New Delhi, India during the years 2009-2017, were included. On multivariate analysis, ECE was significantly associated with depth of invasion >10 mm and tumour deposit size >5 mm. Among the node-positive group, patients without ECE had a 5-year disease-free survival (DFS) and 5-year overall survival (OS) advantage over ECE-positive patients of 7.8% (63.8% vs. 56.0%) and 16.5% (87.2% vs. 70.7%), respectively. For patients with ECE, the hazard ratio for DFS and OS was 1.3 (95% confidence interval 0.97-1.75, P = 0.078) and 2.30 (95% confidence interval 1.35-3.92, P = 0.002), respectively. ECE remains one of the strongest predictors of recurrence and survival in oral cancer patients, and despite aggressive adjuvant therapy, distant recurrence is still significantly high.

Isolation & cryo-preservation of human foetal articular chondrocytes.

An attempt to determine the ideal temperature and duration of storage of human foetal chondrocytes yielded highly cellular preparations with no alteration in morphology or loss of viability. Initial digestion with activated papain was followed by incubation in 0.5 per cent collagenase. Trypan blue exclusion test revealed a viability count of 95-99 per cent and radioactive thymidine uptake a corresponding labelling index. On TEM no subcellular damage was evident. The isolated viable chondrocytes were further banked at varying temperatures of +4 degrees, -4 degrees, -30 degrees, -79 degrees and -196 degrees C, in Eagles MEM with 10 per cent dimethyl sulfoxide. Post storage morphology and viability of these cells, thawed after durations of 20 h, 1 wk, 1, 2, 3, 4 and 6 months, were compared with prestorage readings in an attempt to define the ideal temperature for banking. Storage in liquid nitrogen at -196 degrees C demonstrated excellent preservation even at the end of six months with minimal subcellular change. Electron microscopy and labelling index were found to be superior to Trypan blue exclusion test in assessing the stored chondrocytes for retention of their functions.

Amniotic arthroplasty for tuberculosis of the hip. A preliminary clinical study.

Arthroplasty of the hip using an interposed multi-layered cap of amniotic membrane is reported in 28 patients with tuberculous arthritis. The disease had been present from one to seven years, and five patients had multiple discharging sinuses. Follow-up was from 30 to 46 months. Amniotic tissue caused no inflammatory reaction, or obvious rejection, and 25 patients were free of symptoms, with a good range of movement and a stable joint. The three failures were caused by dislocation, fracture of the femoral neck and extra-articular bone formation respectively. Amniotic arthroplasty seems to be capable of providing a painless, mobile and stable joint in patients with tuberculosis of the hip.

En bloc excision and reconstruction with a fibular autograft of giant-cell tumour of the metacarpal--a case report.

A giant-cell tumour involving the diaphysis of the third metacarpal is reported. The unusual site, age, radiographic atypicality and therapeutic options are reviewed.

Allogenic transplantation of ultra-deep frozen osteochondral grafts. An experimental study in dogs.

Twenty dogs were subjected to allotransplantation of the lower end of the femur, frozen at -196 degrees C for 24 hours in liquid nitrogen, and stored at -30 degrees C. Two dogs were sacrificed at monthly intervals, providing the grafted bone for histological study. Eight dogs were excluded from the study following infection or death, resulting in a follow up of only 6 months. Monthly roentgenographic assessment showed bridging of the host-graft unit by the second month, with posterior subluxation of the graft with subsequent malunion. Histologically, exuberant callus was seen around the interface with progressive incorporation of the graft. However, the articular cartilage degenerated after 2 months. Attempts to repair the articular surface with new fibrocartilage were poor in the malunited grafts. The technical problems in surgery including control of infection, sound mechanical fixation and repair of tendons and ligaments must be solved to ensure a successful allograft.

A small-cell osteosarcoma with multiple skeletal metastases.

A 10-year-old child presented with small-cell osteosarcoma of the proximal end of the tibia. Pulmonary metastases appeared after a gap of 9 months of ablative surgery, as did late multiple extra-pulmonary skeletal metastases, including the hand and foot bones after 6 months of pulmonary metastases. These late extrapulmonary metastases appear to be secondary to pulmonary metastases.

Use of tuning fork in diagnostic auscultation of fractures.

This study was conducted on 50 patients in the Central Institute of Orthopaedics, Safdarjung Hospital, New Delhi, from June to October 1985. With the help of a child's stethoscope and a tuning fork of 128 Hz, the sound conducted by an injured limb was compared with that by the uninjured limb. The presence of a fracture reduced or abolished the conduction of sound by a bone. This method allows a quick examination without causing any pain, which is an advantage in an uncooperative patient. It is also reliable in the unconscious. The test is so simple that paramedical staff can use it. The results were correct in 94 per cent of patients and were confirmed by radiological examination whereas clinical diagnosis was correct in only 88 per cent of cases.