A role for the AKT1 potassium channel in plant nutrition.

In plants, potassium serves an essential role as an osmoticum and charge carrier. Its uptake by roots occurs by poorly defined mechanisms. To determine the role of potassium channels in planta, we performed a reverse genetic screen and identified an Arabidopsis thaliana mutant in which the AKT1 channel gene was disrupted. Roots of this mutant lacked inward-rectifying potassium channels and displayed reduced potassium (rubidium-86) uptake. Compared with wild type, mutant plants grew poorly on media with a potassium concentration of 100 micromolar or less. These results and membrane potential measurements suggest that the AKT1 channel mediates potassium uptake from solutions that contain as little as 10 micromolar potassium.

Molecular evidence for transplacental transmission of Theileria equi from carrier mares to their apparently healthy foals.

The intra-erythrocytic parasite Theileria equi is one of two tick-transmitted causative agents of equine piroplasmosis. Piroplasms of T. equi can be transmitted across the equine placenta and once a horse is infected, it appears to remain a lifelong carrier, since anti-theilerial drugs suppress but do not eliminate the parasite. Carrier mares may transmit the organism to their offspring and this may result in abortion or neonatal piroplasmosis, but observations by some researchers suggest that foals may be born as carriers yet remain apparently healthy. Using a T. equi-specific oligonucleotide probe, we have determined that transplacental transmission occurs early in equine foetal development and that carrier mares may give birth to healthy carrier foals. Investigation of parasite levels and the effect of maternal colostrum on the newborn suggests that colostral T. equi antibody may act to suppress parasitaemia in the newborn, reducing the incidence of clinical neonatal piroplasmosis.

Early recovery after hip arthroscopy for femoroacetabular impingement syndrome: a prospective, observational study.

The early post-operative course after hip arthroscopy for femoroacetabular impingement syndrome has not been thoroughly characterized or correlated to factors that may influence recovery. The aim of this study was to report on early pain, function and attitudes towards rehabilitation and to determine predictors of early recovery after hip arthroscopy. Sixty-two patients reported pre-operative pain, iHOT-12 (hip functional score), psychological status and other baseline characteristics. Pain, iHOT-12, hip flexion and several other outcomes were measured through 6 weeks post-operative. Baseline characteristics were correlated with outcomes using univariate and multivariable models. Pain relief started on post-operative day 1 and consistently improved throughout the 6 weeks of follow-up. The average patient's pain was reduced from a pre-operative level of 5/10 to 2/10 by 6 weeks post-operative. Similarly, iHOT-12 improved from 33/100 to 57/100 whereas hip flexion increased by 9° by 6 weeks post-operative. At 2 weeks post-operative, pre-operative anti-inflammatory usage was associated with greater improvement in pain and swelling; pre-operative opioid usage with poorer patient-reported helpfulness of and adherence to rehabilitation; and higher ASA (American Society of Anesthesiologists) score and lower procedure time with improvement of the pre-operative pain complaint. At 6 weeks, greater depression was associated with lower post-operative pain reduction but greater pre-operative pain complaint improvement. Continuous passive motion usage was associated with increased hip flexion. Pain improved from pre-operative by Day 1 after hip arthroscopy, and early functional improvements were seen by 6 weeks post-operative. Pre-operative anti-inflammatory and opioid usage, depression, race, ASA score, procedure time and continuous passive motion usage were significantly associated with study outcomes.

Immunotherapy of advanced malignancy by direct gene transfer of an interleukin-2 DNA/DMRIE/DOPE lipid complex: phase I/II experience.

PURPOSE: We have completed a phase I study, followed by three phase I/II studies, in patients with metastatic melanoma, renal cell carcinoma (RCC), and sarcoma in order to evaluate the safety, toxicity, and antitumor activity of Leuvectin (Vical Inc, San Diego, CA), a gene transfer product containing a plasmid encoding human interleukin (IL)-2 formulated with the cationic lipid 1, 2-dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide/dioleyl-phosphatidyl-ethanolamine (DMRIE/DOPE) and administered intratumorally. PATIENTS AND METHODS: Twenty-four patients were treated in the phase I study. Leuvectin doses were 10 microg, 30 microg, or 300 microg weekly for 6 weeks. In three subsequent phase I/II studies, a total of 52 patients (18 with melanoma, 17 with RCC, and 17 with sarcoma) were treated with further escalating doses of Leuvectin: 300 microg twice a week for 3 weeks, 750 microg weekly for 6 weeks, and 1,500 microg weekly for 6 weeks. RESULTS: There were no drug-related grade 4 toxicities and only one grade 3 toxicity, but the majority of patients experienced mild constitutional symptoms after treatment. In the phase I/II studies, 45 patients were assessable for response (14 with RCC, 16 with melanoma, and 15 with sarcoma). Two patients with RCC and one with melanoma have achieved partial responses lasting from 16 to 19 months and continuing. In addition, two RCC, three melanoma, and six sarcoma patients had stable disease lasting from 3 to 18 months and continuing. The plasmid was detected by polymerase chain reaction assay in the posttreatment samples of 29 of 46 evaluated patients. Immunohistochemistry studies on serial biopsy specimens showed increased IL-2 expression and CD8(+) infiltration after treatment in the tumor samples of several patients (12 and 16, respectively). CONCLUSION: Direct intratumoral injection of Leuvectin is a safe and possibly effective immunotherapeutic approach in the treatment of certain tumor types.

Potassium uptake supporting plant growth in the absence of AKT1 channel activity: Inhibition by ammonium and stimulation by sodium.

A transferred-DNA insertion mutant of Arabidopsis that lacks AKT1 inward-rectifying K+ channel activity in root cells was obtained previously by a reverse-genetic strategy, enabling a dissection of the K+-uptake apparatus of the root into AKT1 and non-AKT1 components. Membrane potential measurements in root cells demonstrated that the AKT1 component of the wild-type K+ permeability was between 55 and 63% when external [K+] was between 10 and 1,000 microM, and NH4+ was absent. NH4+ specifically inhibited the non-AKT1 component, apparently by competing for K+ binding sites on the transporter(s). This inhibition by NH4+ had significant consequences for akt1 plants: K+ permeability, 86Rb+ fluxes into roots, seed germination, and seedling growth rate of the mutant were each similarly inhibited by NH4+. Wild-type plants were much more resistant to NH4+. Thus, AKT1 channels conduct the K+ influx necessary for the growth of Arabidopsis embryos and seedlings in conditions that block the non-AKT1 mechanism. In contrast to the effects of NH4+, Na+ and H+ significantly stimulated the non-AKT1 portion of the K+ permeability. Stimulation of akt1 growth rate by Na+, a predicted consequence of the previous result, was observed when external [K+] was 10 microM. Collectively, these results indicate that the AKT1 channel is an important component of the K+ uptake apparatus supporting growth, even in the "high-affinity" range of K+ concentrations. In the absence of AKT1 channel activity, an NH4+-sensitive, Na+/H+-stimulated mechanism can suffice.

Ultrasound appearance of synovial osteochondromatosis of the shoulder.

This case report demonstrates the ultrasound appearance of synovial osteochondromatosis of the shoulder, along with plain film radiographic, computed tomographic, and pathologic correlation. Because of recent renewed interest in the use of ultrasonography in the evaluation of musculoskeletal disease, such as rotator cuff arthropathy or suspected intra-articular loose body, opportunities to diagnose synovial osteochondromatosis with this modality have become more frequent.

Current applications of duplex and color Doppler ultrasound imaging: abdomen.

Duplex ultrasound studies and color Doppler imaging have substantially enhanced the diagnostic capabilities of abdominal ultrasonography. The status of the flow in the major abdominal vessels is routinely obtainable along with anatomic information about the organs that they supply. The current applications of duplex and color Doppler imaging in evaluating the hepatic vascular system, hepatic transplants, aorta, splanchnic arterial system, renal artery and vein, renal transplants, and penile arterial system are reviewed. The indications for and limitations of these examinations, as well as the potential future uses, are discussed.

Current applications of duplex and color Doppler ultrasound imaging: carotid and peripheral vascular system.

Recent advances in ultrasound technology have made possible the development of diagnostic instruments that combine cross-sectional imaging and Doppler analysis. These instruments have expanded the role of diagnostic ultrasonography to the assessment of carotid and peripheral vascular disease. The current applications of duplex Doppler and color Doppler imaging in evaluating the extracranial carotid arteries, vertebral artery, peripheral venous system, and peripheral arterial system are reviewed. The indications for and limitations of these examinations, as well as the potential future uses, are discussed.

Radiofrequency treatment of hepatic neoplasms in patients with permanent pacemakers.

Clinicians who provide care for patients with implantable devices for rhythm management, ie, pacemakers and internal cardioverter defibrillators, must be aware of sources of interference that could affect device function. Intracardiac radiofrequency is a recognized source of potential interference. However, radiofrequency to extracardiac sites that are relatively close to the implanted device has not been investigated thoroughly. We present 2 patients with permanent pacemakers undergoing intrahepatic radiofrequency for the treatment of metastatic disease. No interference was documented in either patient. Additional in vitro and in vivo studies are needed to determine definite clinical guidelines for such patients.

Mutation analysis of 28 Gaucher disease patients: the Australasian experience.

Gaucher disease is the most common lysosomal storage disease. It is an autosomal recessive disorder that results from a deficiency of beta-glucocerebrosidase. Three clinical phenotypes have been described: non-neuronopathic, acute neuronopathic, and subacute neuronopathic. Genomic DNA from 28 Australasian patients of diverse ethnic origin with Gaucher disease was screened for 3 common mutations (1226G, 1448C and 84GG) using the amplification refractory mutation system (ARMS), and one uncommon mutation (1504T) by restriction enzyme digestion. Thirty-eight of the 56 independent alleles in these patients were characterized, with 1448C present in 42% and 1226G in 28% of the alleles. The 1226G mutation was associated only with the non-neuronopathic phenotype and 7 of the 15 patients who carried the 1448C mutation developed neuronopathic disease. Three infants who died in the neonatal period following a rapidly progressive neurodegenerative course carried no identifiable mutations. The 84GG mutation was carried by 2 Jewish patients and 1504T was present in one patient. It is now possible to rapidly identify the common Gaucher mutations using ARMS and restriction enzyme digestion, and our findings confirm the heterogeneity of mutations in Gaucher disease. It is also possible to predict in part the phenotypic outcome when screening patients for these mutations. We consider mutation analysis to be of most use in prenatal diagnosis and for carrier detection within affected families.

Fas-induced apoptosis of glioma cells is associated with down-regulation of the hSCO1 protein, a subunit of complex IV.

ApoI/Fas belongs to the tumor necrosis factor receptor (TNFR) superfamily and mediates cell death in various cell types. Earlier studies from this laboratory have shown that Fas-mediated cell death of glioma cells occur, in part, through the production of reactive oxygen species (ROS). To further dissect the molecular mechanisms that are involved in Fas-induced cell death, we compared gene expression between Fas-treated and saline-treated human neuroglioma H4 cells by using the technique of mRNA differential display. This approach led to the identification of hSCO1, a component of the inner mitochondrial membrane, which is required for the correct assembly, and catalytic function of cytochrome-c oxidase, as a Fas down-regulated gene. The decrease in hSCO1 mRNA expression was time-dependent, becoming most prominent after 4 h of Fas-treatment. Morphological changes observed by confocal microscopy revealed that after 4 h of Fas-treatment, the cells undergo membrane blebbing and early formation of apoptotic bodies. These observations are discussed in terms of their support for an important role of mitochondrial events in Fas-induced apoptosis.

Malignant germ cell tumors of the mediastinum.

A review of 56 cases of primary malignant germ cell tumors of the mediastinum revealed that, as with benign teratomas, the tumors occurred in young adults (mean age 29 years) but that the sex distribution differed (86% male and 14% female). A single germ cell element was found in 37 (66%) of the tumors, and various combinations were present in the remaining 19 (34%). The tumors were classified among five recognized types of germ cell tissues. There were 24 seminomas (22 pure and two with mature teratomas), 17 embryonal carcinomas (nine pure and eight with mixtures), five teratomas, seven choriocarcinomas (three pure and four with mixtures), and three pure yolk sac tumors. Most (86%) of the patients were symptomatic at the initial examination, with chest pain, cough, and loss of weight being the most frequent presenting symptoms. The standard posteroanterior and lateral roentgenograms were the most helpful diagnostic tool, showing evidence of an anterior mediastinal mass in 53 patients. The diagnosis was established by surgical exploration of the mediastinum or by biopsy of a lymph node in 55 patients. Of the 55, 24 (43.6%) had complete resection of the tumor and 31 (56.4%) had incomplete resection or biopsy alone. The overall prognosis for mediastinal germ cell tumors is poor, partly because the tumors are far advanced at the time of diagnosis but also because some of the tumors that contain embryonal cell carcinoma, choriocarcinoma, and yolk sac elements are very aggressive. Factors that were prognostic in patients with seminoma--such as age, presence of the superior vena caval syndrome, lymphadenopathy, evidence of hilar disease on the chest roentgenogram, and resectability--were not predictive in patients with other types of malignant germ cell tumors. Although aggressive combination chemotherapy may represent a significant treatment modality for nonseminomatous mediastinal tumors, the present study spanned many years in which no chemotherapy was available. Patients in the later years of the study received combination chemotherapy with various treatment regimens. No conclusions concerning specific chemotherapy, therefore, can be derived from this study.

Benign teratomas of the mediastinum.

Approximately 8% of all mediastinal tumors are benign teratomas. We reviewed 86 cases of benign teratoma seen at the Mayo Clinic from 1930 through 1981. The mean age of the patients was 28 years and the sex distribution was approximately equal. The most common symptoms were chest, back, or shoulder pain, dyspnea, and cough, but 36% were asymptomatic at the time of presentation. Chest roentgenograms showed a well-circumscribed anterior mediastinal mass which often protruded into one lung field. Detectable calcification was observed in 22 patients: a calcified tumor wall in seven, bone or teeth in the mediastinum of seven, and nonspecific calcifications in eight. Surgical excision remains the best means of diagnosing and treating this benign tumor. Though the tumors are histologically benign, they may present difficult surgical problems because of the vital structures involved. Since 1952 there has been a change in the clinical presentation of patients with this entity: More patients are asymptomatic and have smaller tumors and fewer complications than prior to 1952.

Time-course variations in tyrosine hydroxylase activity in the rat locus coeruleus after electrolytic destruction of the nuclei raphe dorsalis or raphe centralis.

Time-course variations in tyrosine hydroxylase activity were measured in the locus coeruleus of the albino rat after electrolytic coagulation of either the nucleus raphe dorsalis or the nucleus raphe centralis. Highly significant increases were measured at 4 days after lesioning of the raphe dorsalis (30.33%) and the raphe centralis (81.55%) compared with control values, whereas the activity in groups A9 and A10 was unchanged at this time-point. In conjunction with other experimental evidences, an hypothesis is proposed that the catecholaminergic neurons located in the locus coeruleus are directly and/or indirectly controlled by the serotonin-containing neurons located in the anterior raphe system nuclei.

The efferent connections of the nucleus raphe centralis superior in the rat as revealed by radioautography.

By chronically implanting a glass micropipette filled with tritiated leucine in the raphe centralis superior of the rat, the projection of this nucleus was traced by radioautography. The majority of the ascending projections were located within the ventral tegmental area and, further rostrally, the median forebrain bundle. Along the course of this bundle numerous fibers branched successively into the mammillary peduncle, the fasciculus retroflexus, the stria medullaris, the fornix and the cingulum. The most significant projections included the ones to the interpeduncular nucleus, the mammillary bodies, the habenular nuclei and the hippocampus. No projections were detected in the striatum, the cortex piriformis or the amygdala. Descending projections diffused to the pontine reticular formation and central gray through the medial and the dorsal longitudinal bundles. In addition widespread projections were also seen in nuclei located near the raphe centralis superior: raphe nuclei, dorsal and ventral tegmental nuclei.

MR angiographic and sonographic indications for endarterectomy.

PURPOSE: Our objective was to determine whether appropriate criteria could be developed for performing an endarterectomy on the basis of sonographic and MR angiographic findings. METHODS: Fifty patients were examined prospectively with sonography, MR angiography, and conventional angiography. All three imaging studies were performed within 2 weeks of one another, and conventional angiography served as the reference standard. RESULTS: All 10 carotid occlusions were detected with sonography and MR angiography. Sonography accurately showed flow in two arteries, and MR angiography showed flow in one of three nearly occluded arteries with extremely slow flow. Multislab three-dimensional time-of-flight MR angiographic sequences underestimated the degree of stenosis in 12 arteries, and in two cases this resulted from high T1 signal within the atherosclerotic plaque. With conventional angiography as the reference standard for 70% to 99% stenosis, sonography had a sensitivity of 96%, a specificity of 91%, and a positive predictive value of 90%, while concordant sonographic findings and the presence of a signal void on multislab 3-D time-of-flight sequences had a sensitivity of 72%, a specificity of 98%, and a positive predictive value of 97%. CONCLUSION: Endarterectomy performed on the basis of sonographic findings of 70% to 99% stenosis and of a signal void on multislab 3-D time-of-flight MR angiographic sequences is appropriate.

Reliability of water proton chemical shift temperature calibration for focused ultrasound ablation therapy.

Our purpose in this work was to assess the reliability of the calibration coefficient for magnetic resonance water proton chemical shift temperature mapping. Over a six month period, the calibration coefficient was measured 15 times in several different phantoms. A highly linear relationship between water proton chemical shift and temperature change was found. The average temperature calibration coefficient determined from all studies was 0.009+/-0.001 ppm/degrees C. Four of the 15 studies were conducted on the same day using the same phantom. The average temperature calibration coefficient of these four studies was 0.0096+/-0.0001 ppm/degrees C.

Factors affecting clinical outcome following endoscopic perforator vein ablation.

BACKGROUND: Despite good outcomes reported with minimally invasive, subfascial endoscopic perforator surgery (SEPS), some patients demonstrate poor healing or recurrence of venous ulcers. The goal of this study was to identify factors that lead to failure of SEPS. METHODS: Forty-eight consecutive patients who had undergone 57 SEPS procedures were analyzed. Mean follow-up was 17 +/- 2 months (range 2 weeks to 52 months). RESULTS: All active ulcers (n = 22) at the time of surgery healed in an average of 99 +/- 37 days (range 11 to 670). Eight limbs had poor healing of their ulcer (>40 days); five (9%) new/recurrent ulcers developed postoperatively. Deep venous obstruction was associated with delayed ulcer healing (316 +/- 171 versus 51 +/- 14 days, P < 0.01) and ulcer recurrence (P < 0.0001). Poor ulcer healing and recurrence were not associated with lipodermatosclerosis, edema, ulcer duration >3 months, or previous recurrences. Ulcer size >2 cm (P < 0.05) and combined ilio-femoral and popliteal/tibial reflux were associated with poor ulcer healing (P < 0.05). CONCLUSIONS: SEPS could not prevent recurrent or new ulceration in 9% of limbs. Venous outflow obstruction was associated with ulcer recurrence and prolonged ulcer healing. Multilevel deep venous reflux and ulcer size >2 cm were also associated with delayed healing.

Intraoperative color-flow duplex ultrasonography following carotid endarterectomy.

Intraoperative color-flow duplex ultrasonography represents the state-of-the-art imaging following carotid endarterectomy. At a reasonable cost and at no risk to the patient, it provides a high-resolution image of the operative site and reproducible hemodynamic velocity measurements. Although significant technical abnormalities have been noted in approximately 10% of patients, important lesions are found in only 5% of common or internal carotid arteries. In our opinion, these common and internal carotid artery defects represent the lesions that should be re-explored and corrected prior to leaving the operating room. The most cost-effective way to utilize intraoperative color-flow duplex ultrasonography is to arrange a 15- to 20-minute visit to the operating room by a technologist bringing a duplex scanner from the vascular laboratory or department of radiology. If the surgeon is not experienced in performing duplex ultrasonography, the test should be performed in conjunction with another physician, surgeon, or radiologist who specializes in ultrasound. In our experience, color-flow duplex ultrasonography enhances endarterectomy technique and consequently minimizes postoperative strokes and restenosis.

Isolation of a South African vector-specific strain of Babesia canis.

A South African strain of Babesia canis parasites was isolated and shown to be vector-specific to only one of the two vectors in the region, Haemaphysalis leachi. This tick was found to transmit the parasite in its adult instar. When infected as larvae, the ticks would not transmit in the proceeding nymphal instar. The vector-specific strain was named the 'Thomas strain' after one of the dogs involved in isolating it. A survey revealed a prevalence of > 50% of this strain in four widely separated areas of the country. Rhipicephalus sanguineus, which transmits B. canis vogeli elsewhere, has not been shown to be a vector of the South African strain of B. canis.