Aggregation of Proteins and its Prevention by Carbohydrate Excipients: Albumins and γ-Globulin

Moisture-induced (2–10 μL added to 10mg) aggregation of solid-state albumin and γ-globulin was investigated by incubation at 37°C for 24 h. The insoluble aggregates were centrifuged from a reconstituted solution, dissolved in a solution containing denaturant and reducing agent, and analysed by a Bio-Rad protein assay kit. Of the three albumins used, maximum aggregation (8·2%) was observed with bovine serum albumin that was essentially fatty-acid free. The maximum aggregation observed with γ-globulin was 70%. A bell-shaped curve for percent aggregation was observed with increasing moisture content and was especially prominent for bovine serum albumin. When mixed with carbohydrate excipients in a 1:1 ratio, aggregation was reduced for both bovine serum albumin and γ-globulin by all four of the following excipients used: Emdex, dextrose, trehalose and hydroxypropyl β-cyclodextrin. For bovine serum albumin, the aggregation was reduced about sixfold, with Emdex being the most effective excipient. The likely mechanism of the resulting aggregation was covalent linkages formed due to intermolecular thiol disulphide interchange.     

Identification of two novel mutations in non-Jewish factor XI deficiency

Summary. We have studied two heterozygous unrelated CRM non-Jewish FXI-deficient patients. Neither of the patients carries a previously described mutation. Their FXI genes were screened by SSCP analysis following PCR amplification of each exon and the flanking intronic sequences. DNA fragments showing aberrant mobility were cloned and sequenced. The following mutations were identified: in case 1, a T to G transition in exon 12 results in the substitution of Phe-442 by Val (FXI-F442V); in case 2 a C to A transition in exon 5 results in the substitution of Cys-128 by a nonsense codon (FXI-C128X). The missense mutation results in a substitution within the protease domain of FXI. Molecular modelling locates this residue in a structurally conserved region of the protease domain and the amino acid substitution may therefore interfere with either chain folding and subsequent secretion or the stability of the protein in plasma. We conclude that the mutations which we have identified are responsible for the inherited abnormality in these patients.     

EFFECT OF ACUTE AND CHRONIC NEUROLEPTIC THERAPY ON SERUM PROLACTIN LEVELS IN MEN AND WOMEN OF DIFFERENT AGE GROUPS

A single fasting level of serum prolactin was measured in each of sixty control subjects and eighty-three psychiatric patients of both sexes who had been on neuroleptic therapy for 2-4 weeks (acute treatment) or at least 5 years (chronic treatment) and who were aged either 17-45 or 48-85 years. All groups of patients had significantly higher mean prolactin levels than controls. Gender, age group of women, and exposure to acute or chronic treatment were significant variables determining the magnitude of neuroleptic-induced elevation of prolactin. In some of the groups, dose, duration of chronic therapy, and concomitant administration of anticholinergic drugs also influenced prolactin levels. Whereas all acutely treated women had prolactin values above the control range, one out of twelve (8.3%) of the women aged 17-45 years and six out of fourteen (42.9%) of the women aged 48-85 years who were under chronic treatment had normal values. Normal prolactin levels were also found in five out of sixteen (31.2%) of the acutely treated and nine out of twenty-four (37.5%) of the chronically treated men aged 17-85 years.     

Unifocal Langerhans’cell histiocytosis (eosinophilic granuloma) resembling Ewing's sarcoma

In a 9 year old boy, a destructive lesion in the diaphysis of the right femur was wrongly diagnosed as a Ewing's sarcoma on the basis of the radiologic findings and fine needle aspiration cytology report. The clinical and radiologic picture was suggestive of Ewing's sarcoma, but an open biopsy of the lesion revealed a histopathological picture of eosinophilic granuloma. A brief review of the literature is given, together with discussion on the differential diagnosis of a mid shaft femoral lesion in young patients.     

Role of videofluorography in assessing functional abnormalities in patients of head and neck cancer treated with chemoradiotherapy

Aim: The major toxicity following treatment for head neck cancer is swallowing dysfunction which can be easily assessed by videofluorography (VFG), allowing documentation of the site and extent of abnormality thereby facilitating directed management. Methods: Between October 2003 and January 2007, 56 patients with locally advanced head and neck cancer were treated by an accelerated radiotherapy schedule with concurrent weekly cisplatin chemotherapy. Three months following treatment, these patients were locally disease free clinically, but complained of varying degrees of dysphagia and were subjected to a VFG evaluation. Results: This group comprised 52 men and four women with a median age of 56 years. The primary site distribution was: oral cavity (9), oropharynx (22), larynx (19), hypopharynx (5) and unknown primary (1). Swallowing function abnormalities in the form of structural displacement and temporal delays were documented and recorded as weakness of the tongue musculature (n = 6), palatal kink (n = 8), premature leak into the oropharynx (n = 20), impaired hyoid elevation (n = 23), impaired epiglottic tilt (n = 26), unilateral pharyngeal wall impairment (n = 16), residuum in vallecula or pyriform fossa (n = 30), aspiration in trachea (n = 29) and loss of nasopharyngeal seal (n = 7). Multiple abnormalities of different sub‐sites were seen in each patient. Conclusion: VFG can document dysmotility disorders of upper aero‐digestive tract like dysfunction of the base of tongue, larynx and pharyngeal musculature leading to stasis of the bolus and vallecular residuum, epiglottis dysmotility resulting in silent aspirations, and inadequate nasopharyngeal seal leading to nasal regurgitation. A clinical correlation alongwith quantification of VFG findings is required.     

Skeletal maturity in Pakistani children

Skeletal maturity in 750 normal Pakistani children (400 males, 350 females) aged 1–18 y was determined by the Greulich–Pyle atlas system. Male children during first year and female children during first 2 y of life matured in conformity with Greulich–Pyle standards. After that period mean bone ages were lower than the American standards up to 15 y in males and 13 y in females (at or around puberty), which may be due to malnutrition, ill health or other environmental factors. After puberty bone ages were higher than the American standards indicating earlier maturity in Pakistani than Western children. Hence for the proper evaluation of skeletal age in a given region, a longitudinal study on individuals in that region to establish normal standards is necessary.     

A COMPARISON OF OLDER LONGSTAY PSYCHIATRIC AND LEARNING DISABILITY INPATIENTS USING THE HEALTH OF THE NATION OUTCOME SCALES

Objective. With increasing longevity, the number of elderly patients with psychiatric or learning disabilities is likely to increase. The degree of overlap of symptoms and needs of these two groups of patients, which may allow for their care within the same service, was examined. Design. Twenty-six longstay, elderly (>65 years) psychiatric patients resident in psychogeriatric wards of a psychiatric hospital were compared with 23 longstay, elderly patients and 40 longstay patients aged 50–65 years, both resident in the wards of a specialist hospital for learning disabilities. The instrument used was the Health of the Nation Outcome Scales (HoNOS). Result. On the HoNOS, the elderly psychiatric patients scored significantly higher for problems with mood, relationships and occupation/activities. There were no significant differences for any of the scales rated between the 50–65 and >65 years old patients with learning disabilities. Conclusion. The similarities between the three groups of patients would suggest that for some patients the same services may be utilized. This could reduce the cost of the care in the community and entail more economical use of the facilities and staff. The HoNOS proved to be a concise and simple instrument, which could become a useful tool in monitoring the outcome of healthcare in longstay patients. © 1997 John Wiley & Sons, Ltd.