Immunocyte Ca2+ influx system mediated by LTRPC2.

We characterized an activation mechanism of the human LTRPC2 protein, a member of the transient receptor potential family of ion channels, and demonstrated that LTRPC2 mediates Ca2+ influx into immunocytes. Intracellular pyrimidine nucleotides, adenosine 5'-diphosphoribose (ADPR), and nicotinamide adenine dinucleotide (NAD), directly activated LTRPC2, which functioned as a Ca2+-permeable nonselective cation channel and enabled Ca2+ influx into cells. This activation was suppressed by intracellular adenosine triphosphate. These results reveal that ADPR and NAD act as intracellular messengers and may have an important role in Ca2+ influx by activating LTRPC2 in immunocytes.

The role, responsibilities and status of the clinical medical physicist in AFOMP.

This document is the first of a series of policy statements being issued by the Asia-Oceania Federation of Organizations for Medical Physics (AFOMP). The document was developed by the AFOMP Professional Development Committee (PDC) and was endorsed for official release by AFOMP Council in 2006. The main purpose of the document was to give guidance to AFOMP member organizations on the role and responsibilities of clinical medical physicists. A definition of clinical medical physicist has also been provided. This document discusses the following topics: professional aspects of education and training; responsibilities of the clinical medical physicist; status and organization of the clinical medical physics service and the need for clinical medical physics service.

Medical physics aspects of cancer care in the Asia Pacific region: 2014 survey results.

It was the aim of this work to assess and track the workload, working conditions and professional recognition of radiation oncology medical physicists (ROMPs) in the Asia Pacific region over time. In this third survey since 2008, a structured questionnaire was mailed in 2014 to 22 senior medical physicists representing 23 countries. As in previous surveys the questionnaire covered seven themes: 1 education, training and professional certification, 2 staffing, 3 typical tasks, 4 professional organisations, 5 resources, 6 research and teaching, and 7 job satisfaction. The response rate of 100% is a result of performing a survey through a network, which allows easy follow-up. The replies cover 4841 ROMPs in 23 countries. Compared to 2008, the number of medical physicists in many countries has doubled. However, the number of experienced ROMPs compared to the overall workforce is still small, especially in low and middle income countries. The increase in staff is matched by a similar increase in the number of treatment units over the years. Furthermore, the number of countries using complex techniques (IMRT, IGRT) or installing high end equipment (tomotherapy, robotic linear accelerators) is increasing. Overall, ROMPs still feel generally overworked and the professional recognition, while varying widely, appears to be improving only slightly. Radiation oncology medical physics practice has not changed significantly over the last 6 years in the Asia Pacific Region even if the number of physicists and the number and complexity of treatment techniques and technologies have increased dramatically.

Pathogenesis of substantia nigra lesions following hyperglycemic ischemia: changes in energy metabolites, cerebral blood flow, and morphology of pars reticulata in a rat model of ischemia.

A spectacular spongiotic lesion, symmetrical in distribution and restricted to the pars reticulata of the substantia nigra (SNPR) has recently been described in hyperglycemic rats surviving 1-18 h after a brief period of transient ischemia. The purpose of this study was to clarify the pathogenesis of the lesion. In order to study whether the lesion was due to changes occurring during ischemia, local cerebral blood flow (l-CBF) and energy metabolites were measured in the substantia nigra (SN) and in other brain areas. Furthermore, brains were examined by light and electron microscopy immediately after ischemia and in the early recirculation period. Autoradiographic CBF measurements showed ischemia flow levels in the SN of 30-40% of control, similar in normo- and hyperglycemic rats. Thus, although ischemic, this structure had a considerable amount of residual flow. There was also a corresponding partial preservation of the adenylate energy charge. However, lactate levels were high, and in hyperglycemic subjects they rose to values previously described during status epilepticus (about 25 mumol/g). In hyperglycemic animals, neuronal alterations were consistently present in SNPR by the end of the 10-min period of ischemia. They included clumping of nuclear chromatin and subplasmalemmal clearing of the perikaryon. Some mitochondrial swelling was present in neuronal perikarya and in dendrites. The normal alignment of microtubules in the dendrites was disturbed, but there was no or only slight swelling of the dendrites. Aggregation of synaptic vesicles was a conspicuous finding in axonal terminals, which were also slightly swollen. Otherwise, the axons appeared largely spared. Microvessels looked quite intact. Similar cellular changes were observed in the early recovery period. Dendrites, however, started to swell, and their expansion finally caused the spongiotic appearance of the pars reticulata. The appearance of the dendritic lesions is strongly suggestive of transmitter-mediated ("excitotoxic") damage. However, it seems likely that the marked acidosis is injurious as well. We tentatively conclude that both mechanisms interact to give the final lesion. The results, and those previously obtained in epileptic seizures, suggest that mitochondria of SN neurons and neuronal processes are particularly prone to damage.

Seizure-induced damage to substantia nigra and globus pallidus is accompanied by pronounced intra- and extracellular acidosis.

Status epilepticus of greater than 30-min duration in rats gives rise to a conspicuous lesion in the substantia nigra pars reticulata (SNPR) and globus pallidus (GP). The objective of the present study was to explore whether the lesion, which encompasses necrosis of both neurons and glial cells, is related to intra- and extracellular acidosis. Using the flurothyl model previously described to produce seizures, we assessed regional pH values with the autoradiographic 5,5-dimethyl[2-14C]oxazolidine-2,4-dione technique. Regional pH values were assessed in animals with continuous seizures for 20 and 60 min, as well as in those allowed to recover for 30 and 120 min after seizure periods of 20 or 60 min. In additional animals, changes in extracellular fluid pH (pHe) were measured with ion-selective microelectrodes, and extracellular fluid (ECF) volume was calculated from the diffusion profile for electrophoretically administered tetramethylammonium. In structures such as the neocortex and the hippocampus, which show intense metabolic activation during seizures, status epilepticus of 20- and 60-min duration was accompanied by a reduction of the "composite" tissue pH (pHt) of 0.2-0.3 unit. Recovery of pHt was observed upon termination of seizures. In SNPR and in GP, the acidosis was marked to excessive after 20 and 60 min of seizures (delta pHt approximately 0.6 after 60 min).(ABSTRACT TRUNCATED AT 250 WORDS)

Decrease of glutamate decarboxylase activity in substantia nigra and caudoputamen following transient hyperglycemic ischemia in the rat.

Glutamate decarboxylase (GAD) activity was determined in caudoputamen (CP), substantia nigra (SN), and cerebral cortex (CCX) after 19-22 h of recirculation following 10 min of transient ischemia in hyperglycemic rats, i.e., under the conditions when previously a pronounced nerve cell damage was demonstrated in both CP and SN. The present results demonstrate a decrease of GAD activity in SN by 30% and in CP by 22% and no change in CCX. No statistically significant change in GAD activity could be detected in SN, CP, or CCX 1,4, and 7 days following 10 min of ischemia in normoglycemic animals. The decrease of GAD activity in SN at the time preceding the onset of postischemic seizures suggests that there may be an imbalance between augmented excitatory and decreased inhibitory transmission in SN. We tentatively conclude that this may increase the probability of generalized seizures in the postischemic period following ischemia in hyperglycemic animals.

Accumulation of calcium and loss of potassium in the hippocampus following transient cerebral ischemia: a proton microprobe study.

This study explored (a) whether postischemic accumulation of calcium in hippocampal neurons precedes or occurs pari passu with light microscopical signs of delayed neuronal necrosis, and (b) whether calcium initially accumulates in dendritic domains, presumed to have a high density of agonist-operated calcium channels. Transient ischemia of 10-min duration was induced in rats, and the animals were studied after 1, 2, 3, and 4 days of recovery. We measured total calcium and potassium contents in the stratum oriens, pyramidale, radiatum, and moleculare of the CA1 and CA3 sectors, using particle induced x-ray emission (PIXE) in the proton microprobe mode. The results showed significant accumulation of calcium and loss of potassium after 3 and 4 days of recovery in the CA1 sector, which developed neuronal necrosis, but not in the CA3 sector, which showed only occasional damage. In a few animals, calcium accumulation (and loss of potassium) was observed with no or only mild visible damage, but in the majority of animals the accumulation of calcium correlated to signs of neuronal necrosis. Since calcium accumulation was similar in all strata examined, the results failed to reveal preferential accumulation in dendritic or somal regions. Based on our results and those of Dux et al., we emphasize the possibility that delayed neuronal death is, at least in part, caused by increased calcium cycling of plasma membranes and gradual calcium overload of mitochondria.

Cyclic guanidines. 17. Novel (N-substituted amino)imidazo[2,1-b]quinazolin-2 ones: water-soluble platelet aggregation inhibitors.

A series of novel 1,2,3,5-tetrahydroimidazo[2,1-b]quinazolin-2-one derivatives substituted with a secondary amino group has been prepared and tested for the activities of inhibiting platelet aggregation in rats in vitro and ex vivo. Most of the compounds were found to be the potent inhibitors of platelet aggregation. Some of the active compounds were soluble in water and effective via iv infusion in rats. Structure-activity relationships have indicated that a lipophilic secondary amino group located at position 6 or 7 contributed to retention of potent activity. Among the compounds studied, 7-piperidino-1,2,3,5-tetrahydroimidazo[2,1-b]quinazolin-2- one (13 g,DN-9693) was the most favorable compound.

Dibasic (amidinoaryl)propanoic acid derivatives as novel blood coagulation factor Xa inhibitors.

Since activated factor X (FXa) is a coagulant enzyme that generates thrombin and participates in both intrinsic and extrinsic coagulation pathways, inhibition of FXa may be more effective than inactivation of thrombin for interrupting blood coagulation. To assess the possible effectiveness of FXa inhibition as an anticoagulant, we designed and synthesized 3-(amidinoaryl)-2-[4-[(3S)-3-pyrrolidinyloxy]phenyl]propanoi c acid derivatives as low molecular weight, nonpeptidic, orally active FXa inhibitors. These derivatives exhibited potent and highly selective anti-FXa activity in vitro and anticoagulant activity on oral administration. The most promising compound, (2S)-2-[4-[[(3S)-1-acetimidoyl-3-pyrrolidinyl]oxy]phenyl]- 3-(7-amidino-2-naphthyl)propanoic acid hydrochloride pentahydrate (4,DX-9065a), inhibited 50% of FXa activity (IC50) at 0.07 microM, doubled plasma recalcification time (PRCT) at 0.5 microM, and significantly prolonged activated partial thromboplastin time (APTT) at a dose of 100 mg/kg on oral administration. In contrast with FXa inhibition, 4 showed no activity against thrombin (IC50 > 2000 microM).

Effects of arginine derivatives on soluble guanylate cyclase from neuroblastoma N1E 115 cells.

The effects of L-arginine (Arg) derivatives on soluble guanylate cyclase from neuroblastoma N1E 115 cells were examined. The Arg derivatives were modified at the -NH2, -COOH, C alpha-proton or guanidino group of Arg. Among the synthesized derivatives, eight compounds, i.e. the 5-(dimethylamino)-1-naphthalenesulfonyl (DNS) ones, especially N-cyclohexyl-2-(N-DNSamino)-5-guanidino-2-methylvaleramide and 1-[2-(N-DNSamino)-2-(2-imino-1,2,3,4,5,6-hexahydropyrimidin- 4-yl)acetyl]- piperidine, were found to inhibit the activity of crude guanylate cyclase in the 105,000 g supernatant fraction of the cell homogenate. The enzyme, partially purified by a column of Chelex 100 Na+, was also inhibited by these eight compounds. The mode of the inhibition was competitive. The Ki values were in the range of 2-8 microM for the enzyme in the 105,000 g supernatant fraction and 3-16 microM for the partially purified enzyme, in the presence of Mg2+ as a metal cofactor. In contrast, a new derivative, methyl 2-amino-5-guanidinovalerate (M Arg ME), as well as the Arg methyl ester (Arg ME) and Arg; were found to enhance the activity of the partially purified guanylate cyclase; KA values of M Arg ME, Arg ME and Arg were approximately 9, 4 and 3 microM respectively. From these results, the free guanidino group including 2-imino-1,2,3,4,5,6-hexahydropyrimidin-4-yl or 2-imino-1,2,3,4,5,6-hexahydropyrimidin-5-yl and modification of the --NH2 residue with a hydrophobic group such as DNS seemed to be essential for inhibition of the guanylate cyclase; however, the guanidino and --NH2 residue of Arg should be free for activation by these Arg derivatives.

Cluster of proliferating cells in rat vomeronasal sensory epithelium.

We investigated the properties of small cells in the vomeronasal sensory epithelium of adult rats. The sensory neurons in the sensory epithelium were stained by antibodies to G(i2alpha) and G(oalpha) in their cell bodies and dendrites, while the small cells, which formed a cluster in the epithelium, were not stained at all. Voltage-activated inward currents were not detected by patch-clamp recordings, but outward currents were induced by the application of voltage step pulses. These results suggest that the small cells are different from the vomeronasal sensory neurons. Bromodeoxyuridine (BrdU) labeling indicated that dividing cells existed in the cluster of small cells.

One-minute wave in body fluid volume change enhanced by postural sway during upright standing.

To analyze how the 1-min oscillation in postural sway and 1-min wave in body fluid volume change contribute to human circulatory homeostasis, several levels of body circumference, foot pressure center, electromyograms, and volumes of the leg, abdomen, and thorax were measured during upright standing for 40 min in 20 healthy young men. Spectral analyses of these parameters revealed that a 1-min rhythm is found in all parameters and that the 1-min wave in body fluid volume changes in the lower leg, which occur in fluid pooling caused by gravity, propagate upward. Muscle pumping in the lower leg triggered by the postural sway was found to increase the power of this 1-min wave. A quantitative analysis of body circumferences disclosed that the 1-min wave in body fluid volume change compensates for gravitational downward fluid shift, with the volume of 6.3 +/- 5.0 ml/cycle at the heart level. We concluded that a coupling mechanism between the 1-min oscillation in postural sway and the upward propagation of 1-min wave in body fluid volume change contributes to maintain systemic blood pressure during upright standing in humans.

Odor discrimination of 'cAMP-' and 'IP3-increasing' odorants at high temperature and at high NaCl concentration in turtle olfactory system.

The ability of the turtle olfactory system to discriminate between various cAMP- and IP3-increasing odorants at high temperature and at high NaCl concentration in the olfactory bulb was examined by the cross-adaptation technique. The degrees of discrimination in high [Na+] solution were similar to those in normal Ringer's solution, suggesting that selectivities of receptors coupled with cAMP- and IP3-dependent pathways are similar to those coupled with both cAMP- and IP3-independent pathways. The mean values of the degree of discrimination among the IP3-increasing odorants were higher than those among the cAMP-increasing odorants at high temperature and at high [Na+] concentration. The degrees of discrimination among the IP3-increasing odorants at 40 degrees C were greater than those at 25 degrees C, while those among the cAMP-increasing odorants at 40 degrees C were similar to those at 25 degrees C, suggesting that the features of the receptors of cAMP-increasing odorants are different from those which respond to IP3-increasing odorants.

Increases in Fos-immunoreactivity after exposure to a combination of two male urinary components in the accessory olfactory bulb of the female rat.

Exposure to either the dialyzed urine preparation (<500 Da) or the remaining substances (>500 Da) did not induce expression of Fos-immunoreactive cells in the mitral/tufted cell layer of the accessory olfactory bulb (AOB), whereas exposure to a mixture of these preparation did induce expression. These results suggest that a combination of low and high molecular weight substances is necessary for the increases in Fos-immunoreactivity in the AOB.

Inositol-1,4,5-trisphosphate accumulation induced by urinary pheromones in female rat vomeronasal epithelium.

The mechanisms involved in pheromone-induced responses in the vomeronasal neurons, especially in mammals, are still unclear. In the present study, we examined the effects of rat urine samples containing various types of pheromones regulating gonadal functions on the accumulation of cAMP and inositol 1,4,5-trisphosphate (IP3) in a vomeronasal membrane preparation from the female Wistar rat. Stimulation of the preparation with forskolin induced cAMP accumulation, but stimulation with urine samples excreted from the male Wistar rat, the female Wistar rat, and the male Donryu rat did not change cAMP levels. These results were consistent with the electrophysiological results showing that dialysis of a high concentration of cAMP into the vomeronasal neuron does not induce currents. Stimulation with the three urine samples induced the accumulation of IP3 in the membrane preparation. These results are consistent with previous electrophysiological results [K. Inamura, M. Kashiwayanagi, K. Kurihara, Inositol-1,4,5-trisphosphate induces responses in receptor neurons in rat vomeronasal sensory slices, Chem. Senses 22 (1997) 93-103; K. Inamura, M. Kashiwayanagi, K. Kurihara, Blockage of urinary responses by inhibitors for IP3-mediated pathway in rat vomeronasal sensory neurons, Neurosci. Lett. 233 (1997) 129-132]. After the treatment with Pertussis toxin (PTX), the male Wistar urine did not induce IP3 accumulation significantly. Application of the male Wistar urine decreased ADP-ribosylation of Gi with PTX, while that of the male Donryu urine decreased ADP-ribosylation of Go. Thus, the present results support a mechanism by which the responses of the rat vomeronasal neurons to urinary pheromones are mediated by IP3, Gi and/or Go.

Accumulation of calcium in substantia nigra lesions induced by status epilepticus. A microprobe analysis.

Status epilepticus of sufficient duration (greater than 30 min) causes a unique lesion of substantia nigra pars reticulata (SNPR), and of globus pallidus (GP). This lesion, which encompasses a pan-necrotic destruction of neurons and glial cells seems to develop during ongoing seizures. We decided to investigate if the lesion is accompanied by net calcium accumulation. Seizures of 20 and 60 min duration were induced by the administration of flurothyl, and the tissue was frozen in situ either at the end of the seizure periods, or after recovery periods of 15 or 120 min. The total calcium and potassium contents of caudoputamen, neocortex, GP and SNPR were measured using particle induced X-ray emission (PIXE) in the microprobe mode. Seizures of 20 min duration did not cause net accumulation of calcium. When the duration of seizures was extended to 60 min the results varied depending on the location. In caudoputamen, which does not incur neuronal damage, no calcium accumulation was observed. In GP and SNPR, such a rise was unequivocally demonstrated, with calcium content increasing to about 150% of controls. The increase in calcium correlated to a decrease in potassium content. It is concluded that epileptic cell death occurs pari passu with accumulation of calcium although it cannot be stated that this accumulation is the cause of the cell death. It is speculated that seizures increase the permeability of the blood-brain barrier to calcium, and that enhanced blood to tissue transfer increases the calcium load of metabolically strained cells.

Blockage of urinary responses by inhibitors for IP3-mediated pathway in rat vomeronasal sensory neurons.

The mammalian vomeronasal system is involved in the effects of urinary chemicals on gonadal functions and sexual behaviors. For example, exposure to urine affects the timing of oestrous cycles in rats. Rat vomeronasal sensory neurons in slice preparation were studied under on-cell patch clamp conditions. We found that urine excreted from male Wistar rats increased impulse frequency in vomeronasal sensory neurons of female Wistar rats. The urinary responses were blocked by an inositol-1,4,5-trisphosphate (IP3)-channel inhibitor (10 microM ruthenium red) or phospholipase C inhibitors (10 microM U-73122 and 1 mM neomycin), suggesting that pheromone-like substances in the urine induce the response in the rat vomeronasal sensory neurons via the IP3-dependent transduction pathway.

An algorithm for mapping the catheter tip position on a fluorograph to the three-dimensional position in magnetic resonance angiography volume data.

This paper proposes an algorithm which maps the position of a catheter tip on a fluorograph to the 3D position in magnetic resonance angiography (MRA) data. This algorithm was assessed for its accuracy. We designed an algorithm consisting of a registration step and a recognition step. The registration step registers MRA and fluorography data using a digital subtraction angiography (DSA) image. The recognition step recognizes the position in the MRA data corresponding to the catheter tip position on a fluorograph. We checked the accuracy of the recognition step by employing an artificial data set consisting of 3D image data (64 x 64 x 64 matrix) and its projection image (92 x 92 matrix) and the accuracy of the registration step with the aid of three of the 3D time-of-flight MRA data sets (256 x 256 matrix and 60 slices) and their projection images in the form of DSA images. The accuracy of the recognition step depended upon that of the registration. When there was no misregistration, all of the mean errors were less than 0.2 mm. The mean errors of the registration step were 0.273 mm and 0.226 mm, respectively, for the longitudinal shift along the X and Y axes, 0.478 degrees, 1.203 degrees and 0.208 degrees, respectively, for the rotation angles around the X, Y and Z axes and 0.020 times for the magnification. The mean image error between the projection image of the registered MRA data and that of the MRA data which were employed as the DSA image was 0.034 mm.

A new capacitive heating applicator for the simultaneous radiohyperthermotherapy of superficial and shallow-seated tumors.

External capacitive heating is the usual method of electromagnetic wave heating, in which the tumor is caught and heated between two opposite applicators. Using a phantom, the authors developed and evaluated the performance of a new capacitive heating applicator designed for simultaneous radiohyperthermotherapy (SRH) in which the electron beam irradiation is provided from above an external capacitive heating applicator for the treatment of superficial and shallow-seated tumors. The trial applicator was constructed to fulfill the following conditions: 1. use of an electrode plate which does not affect the electron beam depth dose, 2. a uniform thickness to maintain flatness of the electron beam, and 3. a cooling function to prevent damage to normal skin tissue and enhance the therapeutic gain factor. This applicator was comprised of a 0.1-mm-thick copper electrode and a 5-mm-thick cooling chamber. The depth of the 80% dose of the new applicator was 21 mm with a 9-MeV electron beam and 36mm with a 15-MeV electron beam, which was comparable to the effect of a conventional irradiation bolus. The temperature distribution produced by the trial applicator was symmetrical on both sides from the center of the applicator. The 50% specific absorption rate region was 6.4 cm wide at a depth of 1 cm from the phantom surface and 2.8 cm wide at a depth of 3 cm. There have been no previous reports on the development of an external capacitive heating applicator designed for the SRH of superficial and shallow-seated tumors; this is the first such report.(ABSTRACT TRUNCATED AT 250 WORDS)

Report of a study using phantom materials, and clinical experience with simultaneous radio-hyperthermotherapy.

Simultaneous radiohyperthermotherapy (SRH) is a combined hyperthermia-radiation therapy in which irradiation is given during heating. Mutual interference between the high energy radiotherapy system (Toshiba LMR-15A) and the 13.56 MHz capacitive heating system (Omron HEH-500C) was tested with phantom materials prior to a clinical trial with SRH. The energy and flatness of irradiation were not affected by the heating system within the range of clinical use. The high energy radiotherapy system did not affect the increase or distribution of temperature during simultaneous treatment. The results of this phantom study indicated that these apparatuses would not produce clinically significant mutual interference during SRH. A clinical trial was performed on a 57-year-old woman with postoperative recurrence of rectal cancer. This is the first reported clinical case treated with true SRH in which external irradiation was administered during mid capacitive heating. Twelve SRH treatments were performed on the recurrent lesion at a frequency of twice a week for six weeks using the apparatuses described above. There was a significant reduction in pain after treatment. The tumor marker carcinoembryonic antigen (CEA) level decreased after treatment. On CT images taken after treatment, the tumor site became a low density area which indicated necrosis. There were no side effects. These results suggest that further clinical study of SRH should be performed to clarify its advantages.