Stage-specific embryonic antigen-4 as a novel marker of ductal cells of human eccrine sweat glands.

BACKGROUND: Different populations of unipotent or multipotent stem cells have been identified in human epidermis and its appendages. It is well documented that these cells maintain tissue homeostasis and actively participate in epidermal regeneration after injury. However, there is no evidence of the presence of pluripotent stem cells in human epidermis. OBJECTIVES: In this study we investigated whether cells positive for embryonic stem cell marker stage-specific embryonic antigen-4 (SSEA-4) are present in adult human epidermis and, if so, whether they are pluripotent and correspond to the population of primitive stem cells. METHODS: The expressions of SSEA-4 and pluripotency transcription factors were analysed using flow cytometry. By means of immunohistochemical staining, we studied the exact localization of these cells in sections of human skin. RESULTS: We show that a population of SSEA-4+ cells is present in human epidermis. In contrast to the commonly accepted belief, the expression of SSEA-4 is not connected with the pluripotent character of isolated cells. We found that these SSEA-4+ cells are localized in the ducts of eccrine sweat glands. CONCLUSIONS: Our results indicate that SSEA-4 is a novel marker identifying the ductal cells of human sweat glands. The surface character of the antigen provides for a simple method of isolating this cell population and suggests applications of SSEA-4 for future cell therapy research.

Activation of new replication foci under conditions of replication stress.

DNA damage, binding of drugs to DNA or a shortage of nucleotides can decrease the rate or completely halt the progress of replication forks. Although the global rate of replication decreases, mammalian cells can respond to replication stress by activating new replication origins. We demonstrate that a moderate level of stress induced by inhibitors of topoisomerase I, commencing in early, mid or late S-phase, induces activation of new sites of replication located within or in the immediate vicinity of the original replication factories; only in early S some of these new sites are also activated at a distance greater than 300 nm. Under high stress levels very few new replication sites are activated; such sites are located within the original replication regions. There is a large variation in cellular response to stress - while in some cells the number of replication sites increases even threefold, it decreases almost twofold in other cells. Replication stress results in a loss of PCNA from replication factories and a twofold increase in nuclear volume. These observations suggest that activation of new replication origins from the pool of dormant origins within replication cluster under conditions of mild stress is generally restricted to the original replication clusters (factories) active at a time of stress initiation, while activation of distant origins and new replication factories is suppressed.

Goiter, cardiovascular and metabolic disorders in patients with acromegaly.

OBJECTIVE: This study evaluated the relationship between selected acromegaly complications such as IGF-1 serum concentrations at diagnosis as well as of controlled and uncontrolled disease. METHODS: A total of 113 acromegaly patients were enrolled to the study and the duration of active and uncontrolled disease was evaluated as a crucial cause of selected complications. RESULTS: Goiter, diabetes, hypercholesterolemia, hypertriglycerydemia, hypertension and ischemic heart disease were diagnosed in 85(75.2 %), 23(20.3 %), 48(51.0 %), 15(13.3 %), 65(57.5 %) and 18(15.9%) patients, respectively. Prevalence of goiter and diabetes was significantly related to the duration of uncontrolled acromegaly (p<0.01) as well as to the prevalence of hypertension and ischaemic heart disease (p0.05). After three years, there was a significant risk of an acromegaly patient being diagnosed at least with one of the above mentioned diseases (p<0.05) and such risk became more significant after four years (p0.05) related to the prevalence of the already mentioned complications. CONCLUSION: The treatment of acromegaly patients should be geared towards fulfilling all criteria for controlled disease, thereby alleviating potential complications and decreasing mortality.

Octreotide lar affects the volume of pituitary adenoma in acromegalic patients.

INTRODUCTION: We studied changes of pituitary adenoma volumes in patients treated with octreotide LAR (SSLAR) over 12 months prior to surgery. MATERIALS AND METHODS: 26 patients (22 female and 4 male, mean age 57.5±15.0 years) were treated with SSLAR: 5 patients with microadenoma - during 6 months, and 21 patients with macroadenoma - during 12 months before surgery. Concentration of hGH and IGF-1 was evaluated at 0, 3, 6 and 12 months, while MRI images were taken at 0, 6 and 12 months prior to surgery. The volume of pituitary adenoma, estimated as that of an ellipsoid, was based on MRI image projections. Basic statistics, Shapiro-Wilk and Wilcoxon tests were applied. RESULTS: Median values of hGH and IGF-1 concentrations prior to treatment were 19.0 ng/ml (IQR=40.7) and 766.4 ng/ml (IQR=787.7), respectively. After 6 and 12 months the median value of hGH concentration decreased to 6.3 ng/ml (IQR=9.0) and 3.45 ng/ml (IQR=3.7), respectively, while the median values of IGF-1 became 535.0 ng/ml (IQR=652.8) and 287.0 (IQR=275.7), respectively. All differences were statistically significant (p<0.05) as compared to basal value. The median volumes of adenomas differed significantly and were estimated at: 1.1 cm (3) (IQR=2.5) prior to treatment, 0.5 cm (3) (IQR=1.5) after 6 months, and 0.35 cm (3) (IQR=2.4) after 12 months of SSLAR administration. CONCLUSION: Treatment of acromegalic patients with somatostatin analogues not only decreases the concentration of hGH and IGF-1, but also appears to decrease the size of the tumour in about 50% of patients treated, leading to general enhancement of the outcome of surgery.

Two rare cases of pituitary metastases from breast and kidney cancers.

Pituitary metastases, though very uncommon, may cause endocrine and neurosurgical problems. The clinical manifestation of such metastases is highly variable. Most of the metastatic pituitary tumours are oligosymptomatic. We report two cases of metastatic pituitary lesions. The first patient, a 52-year old female, with metastatic breast cancer, developed symptomatic anterior pituitary insufficiency. The second patient, a 46-years old female presented with signs and symptoms of pituitary apoplexy and visual impairment due to metastasis from renal cancer. None of them was diagnosed with diabetes insipidus, the most common manifestation related to pituitary metastatic mass.

The unique case of adrenocortical malignant and functioning oncocytic tumour.

UNLABELLED: Adrenocortical oncocytoma is extremely rarely found. Only a little more than thirty cases of adrenal oncocytoma, mainly nonfunctioning and benign, have been reported in the literature. Adrenal mass 150 x 160 x 172 mm in size and enlarged periarterial lymph nodes were found in CT examination performed in 51-year-old male. Main complaints: weight loss, general asthenia and abdominal pain. PHYSICAL EXAMINATION: elevated blood pressure (180/120 mmHg), no features typical of Cushing's syndrome. Abnormal laboratory findings: oral glucose tolerance test revealed diabetes, elevated serum dehydroepiandrosterone-sulfate (1101.9 microg/dl; normal, 59-452), elevated serum cortisol following overnight 1 mg dexamethasone test (5.1 microg/dl; normal, <1.8), increased urinary excretion of 17- hydroxycorticosteroids (18.1 mg/24 h; normal, 2.0-7.0) with pathological response to high-dose dexamethason test (16.6 mg/24). On laparotomy, the lesion was considered unresectable because of evident - confirmed by intraoperative ultrasound - tumour infiltration of the inferior caval vein. The large biopsy specimen was obtained for histological examination in which tumour fulfilled criteria proposed by Bisceglia et al. for adrenocortical oncocytic borderline tumour. On immunohistochemistry, the lesion showed cytoplasmic reaction for cytokeratin, vimentin and synaptophysin. The presented case appears to be the first malignant and functioning adrenocortical oncocytic tumour reported and confirms the complexity of its biology.

Kallmann's syndrome: skeletal and psychological aspects of late diagnosis.

We report a case of Kallmann's syndrome (KS) in a previously untreated 30-year-old Caucasian male, admitted to our Endocrinology Department, presenting with hypogonadotropic hypogonadism and hypoosmia, and reporting a history of rickets in early childhood and a rapid growth pattern. On admission, his main complaints were back-pain and a decreased tolerance to physical exercise. The patient gave no family history of hypogonadism or hypoosmia, and his case was assumed to be sporadic KS. On physical examination hypogonadism (micropenis, small testes, no puberty), hypoosmia and severe scoliosis, kyphosis and chest malformations were recorded. No facial hair growth was found nor past voice braking. His skeletal proportions were eunuchoidal, no mid-line defects were found. This case of KS was identified unusually late. Due to patient's anxiety that his physical appearance might change due to therapy, he was referred to a clinical psychologist who confirmed the patient's self-perception as a male. The risk of further bone malformations, progression of osteoporosis and consecutive pathological fractures were the main indications for commencing treatment. Psychological support was provided. Six months of treatment with low doses of hCG (500 IU given i.m. twice a week) had elevated his testosterone levels but they still remained below the lower values of the normal reference range for males. Additional treatment with vitamin D and calcium supplementation was continued. A certain improvement of bone density score was observed. The patient noted a marked pain relief and he willingly complied with the treatment. After six months of therapy hCG dose was increased to 2000 IU given twice a week. We conclude that even at such a late diagnosis of Kallmann's syndrome in a male who accepts his physical appearance and does not wish any treatment in this respect, hormonal therapy is necessary and should be introduced to reduce significant risk of osteoporosis and bone fractures, and also to offset slowly progressing skeletal malformations which result from the lack of epiphyseal fusion.

The impact of iodine prophylaxis on thyroid 131-iodine uptake in the region of Krakow, Poland.

Iodine prophylaxis was introduced in Poland in 1935. It was interrupted twice, the first time between 1939 and 1947 (due to the Second World War and its aftermath), and then between 1980 and 1986, due to the economical crisis in Poland at that time. A voluntary model of iodine prophylaxis (20 +/- 10 mg/kg of household salt) introduced in Poland in 1986, was followed by a mandatory model, implemented at the beginning of 1997, with 30 +/- 10 mg/kg of household salt. In the early sixties, in our Department of Endocrinology 24-hour iodine uptake test was introduced as a routine procedure for evaluating the thyroid gland. The reference value of this test increased in 1986 after discontinuation of iodine prophylaxis, and decreased in 1998, two years after re-implementation of the mandatory model of iodine prophylaxis. In 167 patients (147 with endemic goiter and 20 with no thyroid disturbances) examined between 1998 and 2000 the mean value of 24-hour iodine uptake was 27.3 +/- 10.4%, as compared to the mean value of 45,7 +/- 6.6 % in 1986. In patients with thyrotoxicosis the mean value of iodine uptake was 41.9% +/- 16.2 in 1999 (no.=614), 42.4% +/- 16.9 in 2000 (no.=644) and 37,7% +/- 17.2 at the beginning of 2001 (no.=328). The mean value of iodine uptake in patients with thyrotoxicosis before implementation of iodine prophylaxis was over 60%. The 24-hour thyroid radioiodine uptake test proved to be a useful indicator of changes of iodine intake in a iodine-deficient population. The data presented in this study confirm the importance and efficacy of the iodine prophylaxis introduced in Poland.

[Pituitary function after transsphenoidal surgery for pituitary adenoma]. / Ocena czynnosci hormonalnej przysadki po operacji gruczolaka z dojscia przez zatoke klinowa.

Nowadays transsphenoidal surgery is the method of choice in most cases of pituitary adenoma treatment, both functioning and non-functioning. This method is considered to be safer than transcranial approach, as lower incidence of complications, particularly hypopituitarism, is observed. The aim of the study was to evaluate the pituitary function after transsphenoidal surgery. 20 patients (mean age 49.3 +/- 13.9 years) were included into the study. The anterior pituitary insufficiency was defined as an inadequate excretory response to metopirone, LH-RH and TRH stimulation. Diabetes insipidus was diagnosed based on clinical symptoms. Hypopituitarism was diagnosed after surgery in 7 patients, 3 cases had adrenal and gonadal insufficiency, 1 patient had insufficiency of the pituitary-thyroid and gonadal axis and 3 subjects panhypopituitarism. Diabetes insipidus was still present in 2 patients, 3 months after surgery. We conclude that transsphenoidal approach in pituitary adenoma surgery is connected with low risk of iatrogenic hypopituitarism.

Plasma endothelin 1/2 levels in healthy blood donors and in hypertensive patients: clinical application.

Normal endothelin 1/2 levels and their correlation with age were evaluated and compared with endothelin 1/2 levels in hypertensive patients. Plasma endothelin 1/2 (ET) levels were measured in healthy blood donors, mostly males, of mean age 36 +/- 8 years (36 subjects), subdivided into three groups: 17-30, 31-40 and above 40 years of age (41-59 yrs). Hypertensive patients (15 subjects) were subdivided into two groups: essential and nephrogenic hypertension. The normal ET levels in the three age groups (means +/- S.D.) were: 0.58 +/- 0.19, 0.62 +/- 0.31, and 0.80 +/- 0.28 fmol/ml, respectively. The average ET level for the whole normal population was 0.66 +/- 0.28 fmol/ml. Only the differences between the mean ET levels in the first and last group were significant (P < 0.05). The difference between the mean ET levels in smokers 0.71 +/- 0.28 fmol/ml (53% of total population) and non-smokers 0.65 +/- 0.28 fmol/ml, women and men, irrespective of age, was not found to be significant. The average ET level in all patients with hypertension (0.91 +/- 0.37 fmol/ml) was significantly higher than the average ET level in blood donors of the same age group (P < 0.05). Although patients with essential hypertension had elevated ET levels compared with control, the difference between the mean ET level in these patients (0.77 +/- 0.24 fmol/ml) and in the corresponsding control group (0.62 +/- 0.31 fmol/ml) was not significant.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma endothelin 1/2 levels in healthy blood donors as measured by RIA--a clinical application.

Normal endothelin 1/2 levels and their correlation with age were evaluated. For clinical application of the endothelin 1/2 RIA test, optimum storage conditions were investigated. Plasma endothelin 1/2 (ET) levels were measured in 36 healthy blood donors, mostly males, of mean age 36 +/- 8 years, subdivided into three age groups: 17-30, 31-40 and above 40 years old. The mean normal ET levels in the three age groups, and corresponding standard deviations, were: 0.58 +/- 0.19, 0.62 +/- 0.31, and 0.80 +/- 0.28 fmol/ml, respectively. The mean ET level for the whole normal population was 0.66 +/- 0.28 fmol/ml. Only differences between mean ET levels for the first and last groups were statistically significant (p < 0.05). Differences between mean ET levels in smokers (53% of total population) and non-smokers, women and men, irrespective of age, were found not to be statistically significant. At this stage of our work, we conclude that other factors than age alone play a role in enhancing ET levels above the age of 41 years. In our study of optimum storage conditions for endothelin 1/2, we found that after one week of storage at -24 degrees C the mean level of ET measured in frozen plasma dropped to 85% of the initial activity, while after the same period the respective decrease in ET activity in frozen extracts was 49%. Over the next two weeks, ET levels in plasma and extracts dropped to 57% and 32% of "zero time" activities, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of calcium and cyclic AMP on amylase release in digitonin-permeabilized parotid gland cells.

Rat parotid cells were permeabilized with digitonin to examine their secretory dynamics. Cells were isolated by a modification of the method previously described by Hootman [1985). J. Biol. Chem. 260, 4186-4194) in which alpha-chymotrypsin was included. The final preparation consisted of approx. 40-60% single cells. The cells were 85-90% viable by trypan blue exclusion and secreted amylase when stimulated with isoproterenol. Digitonin (2 or 5 microM) was sufficient for permeabilization while 2 microM digitonin was somewhat more effective in maintaining cell integrity as indicated by lactate dehydrogenase release. Digitonin had minimal effects on intracellular granules in the whole cell and was, thus, relatively selective. The response of digitonin-permeabilized cells to calcium (without secretagogues) in the incubation medium was monitored by amylase release. For a wide range of applied free calcium concentrations (1 X 10(-7) M to 10(-4) M) a statistically significant increase in amylase secretion was observed. Control cells did not release amylase to a similar extent without secretagogue. Cyclic AMP (50 microM) significantly enhanced amylase secretion from digitonin-treated cells at all concentrations of free calcium tested. Neither calcium nor cyclic AMP alone was sufficient to stimulate maximal amylase release. Our results provide direct evidence for a model in which calcium and cyclic AMP work on separate pathways as interacting regulators of exocytosis.