5-HT-receptor-induced changes of the intracellular cAMP level monitored by a hyperpolarization-activated cation channel

The HvCNG channel from the moth Heliothis virescens is highly sensitive to cAMP concentrations ranging between 0.1 microM and 5 microM. This HvCNG channel was over-expressed in Spodoptera frugiperda (Sf.9) cells to measure endogenous cAMP levels. Hyperpolarization-activated inward currents were measured in the whole-cell patch-clamp configuration with pipettes filled with different cAMP concentrations to calibrate the system. Varying the cAMP concentration between 0 microM and 100 microM in the pipette, the half-maximal activation voltage ( V1/2) was shifted by +28.5+/-1.7 mV. The activation time constant (tau(a)) was used as a parameter for cAMP quantification because it was independent of the expression level of HvCNG channels. tau(a) changed from 1106+/-60 ms at 0 microM cAMP to 265+/-7 ms at a saturating concentration of 1 mM cAMP. A dose-response relationship yielded values of 0.6 microM for the half-maximal cAMP concentration and 1.5 for the Hill coefficient. Activation of endogenous adenylyl cyclases by 50 microM forskolin induced an elevation of the cAMP level by about 1.6+/-0.2 microM. Co-expressions of HvCNG channels in combination with the mouse 5-HT4a- or 5-HT1A- receptors and the corresponding Gs- or Gi-proteins were successful and allowed us to also verify receptor-induced changes of the cAMP level. Stimulation of m5-HT4a-receptors by 0.1 microM 5-HT induced an increase of cAMP of about 4.6+/-1.5 microM, whereas cAMP levels decreased from a control value of 1+/-0.2 microM to 0.41+/-0.1 microM after stimulation of the m5-HT1A-receptors.     

Effects of leg and body position on transcutaneous oxygen measurements in healthy subjects and subjects with peripheral artery disease after lower-extremity arterial revascularization: A pilot study

Transcutaneous oxygen (TcPO(2)) measurements provide a noninvasive, objective determination of the oxygen level at the skin surface. This offers a means of estimating the underlying circulation and tissue oxygenation. The purpose of the pilot study was to measure the TcPO(2) value of the lower extremity of healthy men and women and of patients with peripheral arterial disease (PAD) in 4 different body and leg positions 24 hours after peripheral vascular surgery reconstruction. The specific aim was to determine if lower-extremity TcPO(2) measurements were affected by changes in extremity position in these subject populations. A convenience sample of 4 healthy health care professionals and 4 patients who had peripheral vascular reconstruction surgery 24 hours before the measurements were studied. Subjects were studied in 4 different leg and body positions: supine with legs extended, sitting with legs dependent, a 5 degrees head-up reverse Trendelenburg, and supine with legs elevated 10 in. The Radiometer TCM30 TcPO(2) monitor was used to carry out these measures. Findings revealed a statistically significant difference in TcPO(2) measurements between the 2 groups, with the healthy subjects having a significantly higher TcPO(2) measurement in all extremity positions compared with the revascularized subjects with PAD (P =.02-.05). Significant changes were noted in both the foot temperature (P =.03) and TcPO(2) measurements with extremity positions within the healthy subject group (P =.001). The foot and leg TcPO(2) measurements affect from leg and body position did not reach significance (P =.09) in the subjects with PAD. No change in foot temperature with extremity positioning (P =.42) was noted in the subjects with PAD. This pilot study provides a base in which additional research will be performed with TcPO(2) measurements in both the healthy and revascularized person.     

Familial expansile osteolysis: a morphological, histomorphometric and serological study.

Biopsies from the diseased bones of patients with familial expansile osteolysis (FEO) were examined by light and electron microscopy. Focal concentrations of multinuclear osteoclasts were present, and these contained viral-like microcylindrical inclusions which appeared exclusive to their nuclei. No consistent relationship was found between osteoclast size and the number of osteoclast nuclei containing microcylindrical inclusions. Quantitative histomorphometry showed evidence of increased bone remodelling with high bone cell densities and a decrease of the reversal period in bone remodelling. The lesions contained prominent woven bone and fibrovascular tissue, together with mononuclear cells and adipocytes. Little bone was found in the most radiolucent lesions, which were almost totally occupied by adipocytes and fibrovascular tissue. Serology did not reveal any significant differences between the viral antibody titres of patients and their age- and sex-matched controls. The present study suggests that intranuclear viral-like microcylindrical inclusions of osteoclasts are not a specific feature of Paget's disease, and are found in other disorders of osteoclast function, including pycnodysostosis, osteopetrosis, giant cell tumours, and familial expansile osteolysis.     

Papilliferous keratoameloblastoma

A case of papilliferous keratoameloblastoma is reported which is only the second ever documented. The patient was a 76-yr-old black woman with a large expansile multilocular radiolucency of the body, angle and ramus of the mandible. Histologically the lesion consisted of sheets of cystic follicles filled with necrotic debris and sometimes parakeratin. The vast majority of the follicles were lined by a papilliferous epithelium consisting of large rounded cells with centrally placed nuclei. True papillary projections with cores of connective tissue were also present. The remainder of the follicles were lined by a thin parakeratinising stratified squamous epithelium. Histological features characteristic of ameloblastoma were absent. Final classification of these lesions will have to await the reporting of further cases.