Ric-8A gene deletion or phorbol ester suppresses tumorigenesis in a mouse model of GNAQ(Q209L)-driven melanoma.

The heterotrimeric G protein α subunit oncogenes GNAQ or GNA11 carry Q209X or R183X activating mutations and are present with ~90% frequency in human uveal melanomas. Forced expression of GNAQ/11(Q209L) in melanocytes is sufficient to drive metastatic melanoma in immune-compromised mice. No known drugs directly target these oncogenic G proteins. Ric-8A is the molecular chaperone that selectively folds Gαq/i/13 subunits. Targeting Ric-8A serves as a rational, yet unexplored approach to reduce the functional abundance of oncogenic Gαq/11 in order to blunt cancer signaling. Here, using mouse melanocyte cell graft tumorigenesis models, we determined that Ric-8A genetic ablation attenuated the abundance and melanoma-driving potential of Gαq-Q209L. A new conditional Ric-8A(Flox/Flox); Rosa-CreER(+/)(-) mouse strain was derived and used as a tissue source to culture an immortalized, tamoxifen-inducible Ric-8A knockout melanocyte cell line that required 12-O-tetradecanoylphorbol-13-acetate (TPA, phorbol ester) for growth. The cell line failed to grow tumors when grafted into immune-compromised mice regardless of Ric-8A expression. Stable expression of human GNAQ(Q209L), but not GNAQ(WT) in the cell line promoted TPA-independent cell proliferation, and upon cell grafting in mice, the initiation and robust growth of darkly-pigmented melanoma tumors. Deletion of Ric-8A in GNAQ(Q209L) cells restored TPA-dependent growth, reduced Gαq-Q209L below detectable levels and completely mitigated tumorigenesis from primary or secondary cell line grafts. Interestingly, TPA treatment of cultured GNAQ(Q209L) cells or host animals grafted with GNAQ(Q209L) cells also sharply reduced Gαq-Q209L abundance and tumorigenic capacity. Finally, tumorigenesis initiated from GNAQ(Q209L) cell grafts, followed by host mouse systemic tamoxifen treatment to delete Ric-8A in the grafted cells completely abrogated GNAQ(Q209L)-driven tumor progression unless a stable human RIC-8A transgene was used to rescue the floxed Ric-8A alleles. Our work defines two new rational targets that may be developed as potential uveal melanoma therapies through reduction of Gαq/11-Q209L oncoprotein abundance: (1) Ric-8A inhibition and (2) phorbol ester treatment.

Acute toxicity and antihyperlipidemic activity of rhizome of Tectaria coadunata (Kukkutnakhi): A folklore herb.

BACKGROUND: Tectaria coadunata (Wall. Ex Hook and Grev.) C. Chr (Kukkutnakhi) is a pteridophyte fern which is found in Western Ghats, Kerala Ghats, and Mahendragiri forest region. It is used by many Vaidyas in hyperlipidemic conditions and obesity. AIM: This study aimed to evaluate the acute toxicity and antihyperlipidemic activity of T. coadunata in experimental animals. MATERIALS AND METHODS: Oral acute toxicity study was carried out in female Wistar rats as per OECD 425 guideline. Antihyperlipidemic activity of powder of T. coadunata (540 mg/kg) was carried out in high-fat diet-induced hyperlipidemia in Wistar albino rats. RESULTS: T. coadunata rhizome powder at the dose of 2000 mg/kg did not produce any mortality and toxic effects during acute toxicity study in female rats. Test drug produced highly significant (P < 0.001) reversal in the triglycerides and very-low-density lipoprotein (VLDL)-cholesterol along with nonsignificant decrease in the cholesterol level in rats fed with hyperlipidemic diet. Further, T. coadunata has shown nonsignificant decrease in serum urea, serum glutamic pyruvic transaminase, alkaline phosphatase, and bilirubin direct while statistically significant decrease in bilirubin total in comparison to cholesterol control group. Histopathological study has shown reversal of adverse changes induced by hyperlipidemic diet in heart, liver, and kidney. CONCLUSION: It is concluded that drug is safe up to the dose level of 2000 mg/kg in rats. Rhizome of T. coadunata has shown antihyperlipidemic activity in rats, which suggest its potential role in hyperlipidemia and associated conditions.

Anti-inflammatory activity of root bark and stem bark of Shyonaka.

BACKGROUND: Shyonaka (Oroxylum indicum Vent.; Bignoniaceae) root bark is one of the ingredients of dashamoola (a group of 10 roots), and is used for its anti-inflammatory and analgesic action in a number of compound formulations in Ayurveda. AIM: Ayurvedic Pharmacopoeia of India (API) recommends using the stem bark instead of root bark. MATERIAL AND METHODS: An attempt has been made to study the anti-inflammatory activity of both root bark and stem bark kashaya (decoction) experimentally. CONCLUSION: Results showed significant anti-inflammatory activity of root bark and stem bark decoction.

The median whole-blood donation time in the best-case scenario: findings at one blood center.

BACKGROUND: Whole-blood donation times in a donor population when there are no impediments to blood donation at any point can be used to determine the shortest median whole-blood donation time. This statistic is relevant to blood donors and collection staff. STUDY DESIGN AND METHODS: Small, over-staffed blood drives in August 2004 were evaluated via operation records and supervisor interviews to determine if there were any impediments to whole-blood donation at any point in the process. Four-hundred ninety-two whole-blood donations from 33 blood drives were not impeded and were evaluated for whole-blood donation times. Registration records (entry time) and blood donor records (phlebotomy start and end time) provided three objective time points for the process, the median post-phlebotomy interval at the phlebotomy station was estimated, and 10 minutes was assigned for the refreshment period. RESULTS: The median donation time for the whole-blood donation process was 50 minutes. The 25 to 75 percent range was 46 to 56 minutes, and the 10 to 90 percent range was 42 to 64 minutes. First-time blood donors had blood donation times that were 5 to 6 minutes longer than in repeat blood donors (55.5 min vs. 50 min), and women had slightly longer donation times than men (51 min vs. 49 min). Five percent of the donors had donation times that exceeded 69 minutes. CONCLUSION: The median whole-blood donation time and range under the best circumstances was defined based on the process in place in our blood center in August 2004.

A comparative study of subsyde-CR versus meloxicam in rheumatic disorders.

A prospective, randomised comparative clinical study was conducted in adult patients of either sex presenting with articular and non-articular rheumatic conditions commonly encountered in clinical practice Rheumatoid arthritis, osteo-arthritis, cervical spondylosis, and lumbago/sciatica were the most frequent conditions encountered in both the groups, followed by others like tenosynovitis, frozen shoulder, prolapsed disc, fibrositis, myositis, sprains, strains and so on. The drugs that were employed for therapy were diclofenac in a controlled release formulation employing the DRCM technology (subsyde-CR) and meloxicam in a standard formulation marketed in our country. Both drugs were well tolerated and found to be effective in reducing the signs and symptoms of the disease entities throughout the study period, but subsyde-CR was observed to produce a somewhat greater reduction in signs and symptoms scores that meloxicam, a difference that could be possibly attributed to the greater efficacy of subsyde-CR in non-articular rheumatic conditions. On the basis of the available literature on diclofenac and meloxicam as well as the DRCM technology in formulating subsyde-CR, it is reasonable to conclude that a controlled release formulation of diclofenac based on the DRCM technology offers a safe and effective alternative to other non-steroidal anti-inflammatory drugs such as meloxicam.

Persistence of cerebral blood flow after brain death.

Persistent cerebral blood flow occasionally confounds confirmatory tests for brain death and results in the anguish of delayed diagnosis, unnecessary use of expensive resources, and loss of transplant opportunities. We reviewed the literature to examine the reasons, frequency, and meaning of this problem. We found that this phenomenon occurs: (1) before increasing intracranial pressure completely shuts down flow; (2) in infants with pliable skulls; and with (3) decompressing fractures, (4) ventricular shunts, (5) ineffective deep brain flow, (6) reperfusion, (7) brain herniation, (8) jugular reflux, (9) emissary veins, and (10) pressure injection artifacts. Isolated venous sinus visualization is common (occurring in up to 57%) but represents trivial blood flow and confirms brain death. Arterial flow is much less common (2.6% incidence in our series). Normal flow occurs but is rare. Arterial flow does not exclude brain death, but the diagnosis should be confirmed by repeated studies or other means.

Accuracy of clinical evaluation in the determination of brain death.

BACKGROUND: The accuracy of the clinical diagnosis of brain death has never been established. METHODS: Seventy-one consecutive clinically brain dead patients were studied retrospectively. Inclusion criteria were complete cessation of brain function with profound coma of known cause, complete absence of brain stem reflexes, and apnea, all persisting for a least 6 hours. A formal apnea test with a documented Pco2 of > 60 mm Hg was required. All evaluations were done by experienced neurosurgery or neurology resident or staff physicians. The clinical diagnosis was compared with the results of radionuclide angiography and with the clinical course and final outcome. RESULTS: Seventy patients had no arterial blood flow on radionuclide angiography. One blood flow study was considered to have yielded a false-negative result. No patient recovered or survived. CONCLUSIONS: The clinical diagnosis of brain death is highly reliable when made by experienced examiners using established criteria. In this study, the accuracy was 100%.

Radionuclide angiography as a confirmatory test for brain death: a review of 229 studies in 219 patients.

BACKGROUND: Radionuclide angiography (RA) has been used for many years to confirm the clinical diagnosis of brain death, but quantitative data regarding the technique are unavailable. METHODS: We conducted a retrospective review of 229 RA procedures done in 219 patients. RESULTS: Of 206 patients who had interpretable studies and met clinical criteria for brain death (profound coma, absent brain stem reflexes, and apnea), including 10 patients who had repeated studies, 203 (98.5% sensitivity) had positive studies, and 6 (2.9%) had negative studies. Of nine patients who had interpretable studies and did not meet the clinical criteria for brain death, five (56% specificity) had positive studies and four (44%) had negative studies. CONCLUSIONS: Radionuclide angiography is a sensitive confirmatory test of brain death. The study does not evaluate the posterior fossa circulation, and an occasional patient may have residual brain stem function. The examination is most reliable when the patient is in a deep coma, has no brain stem reflexes, and has failed a formal apnea test.

Pediatric Graves' disease: therapeutic options and experience with radioiodine at the University of Mississippi Medical Center.

BACKGROUND: Pediatric Graves' disease can be life-threatening, and it adversely alters growth and development. Controversies concerning optimal therapy led us to review our 40 pediatric patients treated for Graves' disease from 1988 to 1996 to assess efficiency, efficacy, and safety of current therapy options. METHODS: Diagnosis of Graves' disease required clinical hyperthyroidism with supportive laboratory studies. Patients were given informed choices of therapy, which divided them into three groups. RESULTS: In group 1, 17 patients received antithyroid medications for 0.3 years to 6.0 years. Three required surgical thyroidectomy. Remissions (with or without thyroxine therapy) were achieved after 2 years to 5 years in 11 (65%). In group 2, 15 patients received antithyroid medications for 0.3 years to 5.0 years before receiving radioactive iodine (131I). One also required surgical thyroidectomy. Remissions were achieved after 1 year to 5 years in 10 (67%). In group 3, eight patients received initial 131I therapy. Remissions were achieved within 1 year in 7 (88%). CONCLUSIONS: Our results agree with and expand upon published reports on Graves' disease. Our data support early use of 131I as efficient, effective, and safe therapy for pediatric Graves' disease.

Fracture-dislocation at the thoracolumbar junction in an infant with locked vertebrae. A case report.

STUDY DESIGN: A retrospective, single-patient case report. OBJECTIVES: To report on a 9-month-old infant with fracture-dislocation at the thoracolumbar junction with locking of vertebral bodies, probably with incomplete neurologic injury and partial postoperative recovery. SUMMARY OF BACKGROUND DATA: Fracture-dislocation at the thoracolumbar junction is rare in infants, the upper cervical spine being the level usually affected. Incomplete neurologic injury is uncommon after such a severe shear injury. METHODS: The patient was followed through personal examination and chart review from initial presentation to 6 months after the injury. RESULTS: Open reduction with posterior spinal fusion was followed by partial neurologic recovery and a stable spine over 6-month follow-up. CONCLUSIONS: Fracture-dislocation at the thoracolumbar junction is rare in infancy. This is probably due to unique anatomic and biomechanical features of the spine in infancy.

Two stage reconstruction for the Shepherd's crook deformity in a case of polyostotic fibrous dysplasia.

Polyostotic fibrous dysplasia leads to progressive and disabling deformity involving the proximal femur. Conventional methods of treatment have been ineffective in controlling this problem. Two stage reconstruction was carried out in a case of polyostotic fibrous dysplasia with bilateral shepherd's crook deformity. Bilateral subtrochanteric osteotomies with intramedullary fixation in the first stage and intertrochanteric osteotomies in the second stage with nail plate fixation was done to provide definitive control of the deformity. Bone graft was not used.

Systemic radionuclide therapy with strontium chloride Sr 89 for painful skeletal metastases in prostate and breast cancer.

Systemic radionuclide therapy with strontium chloride Sr 89 is a rediscovered alternative to relieve pain from bony metastases. Although numerous advances have been made in the diagnosis and treatment of cancer, pain remains a serious and debilitating disease complication. An increasing number of clinical trials are reporting satisfactory results with 89Sr-chloride therapy, now available for widespread clinical use. We have treated 11 patients with this radionuclide; of these patients, 8 had excellent to dramatic pain relief and 3 had mild to moderate improvement. Clinical response was based on subjective pain relief, increased mobility, decreased analgesic uptake, and/or improvement in daily activities, including work habits.

Is McMurray's osteotomy obsolete?

A review of the method of performing, advantages, disadvantages of McMurray's displacement osteotomy with regard to treatment of nonunion of transcervical fracture neck femur with viable femoral head was carried out in this study of ten cases, in view of the abandonment of the procedure in favour of angulation osteotomy. Good results obtained in the series attest to the usefulness of McMurray's osteotomy in the difficult problem of nonunion of transcervical fracture neck femur in well selected cases with certain advantages over the angulation osteotomy due to the 'Armchair effect'.

Evaluation of the activity of a number of antimicrobial agents against mycobacteria within mouse macrophages by a radiometric method.

[3H]-uridine was incorporated by Mycobacterium bovis BCG with increasing intensity as the incubation period was increased. Rifampicin and isoniazid inhibited incorporation of the label rapidly. Similar inhibition was seen with M. tuberculosis H37Rv and several clinical isolates of M. tuberculosis both in axenic medium and inside macrophages. Ofloxacin and ciprofloxacin were both inhibitory but clofazimine was not. The combination of rifampicin with either isoniazid or ethambutol produced enhanced killing, but the combination of ethambutol and isoniazid was not synergic. Mycobacterium avium-intracellulare isolates from AIDS patients were less susceptible to rifampicin and were unaffected by isoniazid, ethambutol, clofazimine, ofloxacin and ciprofloxacin. The results obtained by inhibition of [3H]-uridine incorporation by intracellular mycobacteria correlated with conventional in-vitro MICs and was reproducible and rapid; a definitive result was obtainable within seven days.

Bizarre parosteal osteochondroma in the foot (a case report).

A rare case of a bizarre parosteal osteochondroma in the foot is presented. The tumour first described in 1983 by Nore et al is rare in any location and has to be differentiated from a juxta-cortical osteosarcoma and a benign osteosarcoma. Its differentiating characteristics are discussed.

Cholinergic and serotonergic effects on the P3 potential and recent memory.

The effects of scopolamine (anticholinergic) and methysergide (antiserotonergic) on memory and long-latency auditory cognitive evoked potentials (EPs) were tested in 16 normal adults. Recent memory was impaired by both drugs. In contrast, scopolamine, but not methysergide, significantly delayed P3 latency and decreased P3 amplitude. Immediate memory and the earlier EP components (i.e., N1 and P2) were unaffected. The findings support the hypothesis that cholinergic neurons are important in the neuronal networks generating the P3 potential. Serotonergic neurons do not appear to play a major role in the networks that generate the P3.

Central cholinergic systems and the P3 evoked potential.

A cognitive evoked potential, the P3, is commonly altered in dementia states but little is known of the specific neuronal generators that are the source of this potential. Event-related evoked potentials and neuropsychological testing were obtained in six normal subjects during neuropharmacological manipulation of the central cholinergic system. Scopolamine (an anticholinergic) impaired recent memory, prolonged P3 latency and decreased P3 amplitude. These abnormalities were partially reversed by physostigmine (an anticholinesterase). The results imply that the cholinergic system is involved in the generation of the P3 potential.