Mitochondrial sequence analysis for forensic identification using pyrosequencing technology.

Over recent years, requests for mtDNA analysis in the field of forensic medicine have notably increased, and the results of such analyses have proved to be very useful in forensic cases where nuclear DNA analysis cannot be performed. Traditionally, mtDNA has been analyzed by DNA sequencing of the two hypervariable regions, HVI and HVII, in the D-loop. DNA sequence analysis using the conventional Sanger sequencing is very robust but time consuming and labor intensive. By contrast, mtDNA analysis based on the pyrosequencing technology provides fast and accurate results from the human mtDNA present in many types of evidence materials in forensic casework. The assay has been developed to determine polymorphic sites in the mitochondrial D-loop as well as the coding region to further increase the discrimination power of mtDNA analysis. The pyrosequencing technology for analysis of mtDNA polymorphisms has been tested with regard to sensitivity, reproducibility, and success rate when applied to control samples and actual casework materials. The results show that the method is very accurate and sensitive; the results are easily interpreted and provide a high success rate on casework samples. The panel of pyrosequencing reactions for the mtDNA polymorphisms were chosen to result in an optimal discrimination power in relation to the number of bases determined.

Adrenal crisis in active duty service members.

Primary adrenal insufficiency is a chronic, debilitating condition that usually produces a variety of characteristic but non-specific clinical features. Up to 25% of patients present instead with acute life-threatening adrenal crisis, marked by severe hypotension and shock. Recognition of the disease in the chronic indolent phase is critical because adrenal steroid replacement effectively relieves symptoms and prevents the development of most acute crises. To illustrate these points, we describe four case histories in which the manifestations of chronic adrenal insufficiency went unrecognized in active duty service members until they presented with near-fatal adrenal crises. The salient clinical features, diagnosis, and treatment of the disease are also reviewed.

Effects of weight loss on single meal eating behaviour in obese subjects.

In 22 obese patients the eating pattern during a single meal as evaluated by the universal eating monitor VIKTOR was assessed after one year of treatment, initially with VLCD (Nutrilett) followed by combined long term diet, exercise and behavioural modification, underscoring the importance of appropriate meal habits. A mean weight loss from 117 to 101 kg was achieved. Before treatment a decelerated eating curve was found; after treatment this curve changed significantly towards a flatter and more linear shape.

Hypercalcaemia and elevated levels of parathyroid hormone-related protein in cutaneous squamous/basal cell carcinoma.

A patient with mixed squamous/basal cell carcinoma of the skin presented with hypercalcaemia and elevated serum levels of parathyroid hormone-related protein (PTH-rP). The tumour was resected, PTH-rP levels declined and the patient became normocalcaemic. This is the first case to associate squamous cell carcinoma of the skin with hypercalcaemia and significant levels of PTH-rP.

Evidence of endocrine involvement early in the course of human immunodeficiency virus infection.

Adrenal, gonadal, and thyroid function were assessed in 40 asymptomatic subjects in whom infection with the human immunodeficiency virus (HIV) had recently been documented. None of the patients had historical or clinical evidence of endocrine dysfunction. Their mean serum hormone levels were also within the expected ranges, but several differences were noted compared to those of seronegative controls. Basal cortisol, basal aldosterone, and ACTH-stimulated cortisol were significantly lower in the HIV group. One subject (2.5%) had a subnormal cortisol response, and two (5%) had abnormal aldosterone responses to ACTH. PRA tended to be higher, and serum angiotensin-converting enzyme levels somewhat lower in the HIV group. Serum free testosterone was markedly elevated in the HIV patients and was associated with an exaggerated LH response to GnRH, but PRL, estradiol, and basal and peak GnRH-stimulated FSH did not differ between groups. Three subjects (8%) had subclinical hypothyroidism. Serum thyroid hormone levels were normal, but basal T3 was lower in the HIV group compared to control values. While of little immediate clinical importance, many subtle endocrine aberrations are evident very early in the course of HIV infection. These findings obtained in HIV-seropositive subjects without infections or tumors and who were not receiving medical therapy suggest an effect of HIV on each of the endocrine systems examined.

Fine needle aspiration of the thyroid. Use in an average health care facility.

Fine needle aspiration of the thyroid is suggested as an initial diagnostic procedure in the evaluation of thyroid nodules. Some proponents of the procedure, however, believe that fine needle aspiration should be restricted to centers in which each operator obtains at least 500 biopsy specimens annually. Reviewed were 155 biopsy specimens obtained over three and one-half years, and the sensitivity of fine needle aspiration was determined to be 100 percent, the specificity to be 47.4 percent, and the accuracy to be 73 percent. A review of the literature revealed the sensitivity in larger series to range between 92 and 98 percent, specificity to range between 52 and 99.5 percent, and accuracy to range between 56.1 and 91.3 percent. The surgical yield of carcinoma was 64 percent in patients evaluated with fine needle aspiration, whereas the yield was 26 percent in those who underwent surgery without fine needle aspiration. These results appear to justify the use of fine needle aspiration in the evaluation of nodular thyroid disease at average-sized health care centers.

Ectopic thyroid tissue on thallium/technetium parathyroid scan.

The thallium 201/technetium 99m pertechnetate radionuclide study is becoming widely accepted as a means of localizing abnormal or aberrantly located parathyroid tissue. We describe a case in which ectopic retrosternal thyroid tissue appeared as a parathyroid adenoma on thallium-201/technetium-99m pertechnetate scan. Physicians who use this radionuclide study should be aware of the possibility of false-positive images within the mediastinum.