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Purpose: To quantify the two principal forms of hepatic storage iron, diffuse, soluble iron (primarily ferritin), and aggregated, insoluble iron (primarily hemosiderin) using a new MRI method in patients with transfusional iron overload., Materials and Methods: Six healthy volunteers and 20 patients with transfusion-dependent thalassemia syndromes and iron overload were examined. Ferritin- and hemosiderin-like iron were determined based on the measurement of two distinct relaxation parameters: the "reduced" transverse relaxation rate, RR2 , and the "aggregation index," A, using three sets of Carr-Purcell-Meiboom-Gill (CPMG) datasets with different interecho spacings. Agarose phantoms, simulating the relaxation and susceptibility properties of tissue with different concentrations of dispersed (ferritin-like) and aggregated (hemosiderin-like) iron, were used for validation., Results: Both phantom and in vivo human data confirmed that transverse relaxation components associated with the dispersed and aggregated iron could be separated using the two-parameter (RR2 , A) method. The MRI-determined total hepatic storage iron was highly correlated (r = 0.95) with measurements derived from biopsy or biosusceptometry. As total hepatic storage iron increased, the proportion stored as aggregated iron became greater., Conclusion: This method provides a new means for noninvasive MRI determination of the partition of hepatic storage iron between ferritin and hemosiderin in iron overload disorders., (Copyright © 2013 Wiley Periodicals, Inc.)

Purpose: To investigate blood flow and transit time measurement, using the pseudo-random arterial modulation (PRAM)., Materials and Methods: PRAM is based on a pseudo-random sequence of inversions and noninversions of the arterial blood at a labeling plane inferior to the imaging plane. To accomplish this, a pseudo-continuous tagging is used to create inversion or noninversion prepulses before a gradient echo sequence and tested on phantoms and human volunteers., Results: We have shown here that the PRAM technique can measure the velocity profile and the transit time accurately and efficiently both in a phantom and in vivo in a human brain., Conclusion: PRAM does not require separate control and label acquisition as is common in arterial spin labeling (ASL) but rather measures the distribution of transit times to a voxel within one integrated scan. The PRAM method is a model-free approach in measuring transit time distributions, and therefore ultimately should provide more accurate perfusion measurements., (Copyright © 2011 Wiley Periodicals, Inc.)

Purpose: To evaluate the reduced transverse relaxation rate (RR2), a new relaxation index which has been shown recently to be primarily sensitive to intracellular ferritin iron, as a means of detecting short-term changes in myocardial storage iron produced by iron-chelating therapy in transfusion-dependent thalassemia patients., Materials and Methods: A single-breathhold multi-echo fast spin-echo sequence was implemented at 3 Tesla (T) to estimate RR2 by acquiring signal decays with interecho times of 5, 9 and 13 ms. Transfusion-dependent thalassemia patients (N = 8) were examined immediately before suspending iron-chelating therapy for 1 week (Day 0), after a 1-week suspension of chelation (Day 7), and after a 1-week resumption of chelation (Day 14)., Results: The mean percent changes in RR2, R2, and R2* off chelation (between Day 0 and 7) were 11.9 ± 8.9%, 5.4 ± 7.7% and -4.4 ± 25.0%; and, after resuming chelation (between Day 7 and 14), -10.6 ± 13.9%, -8.9 ± 8.0% and -8.5 ± 24.3%, respectively. Significant differences in R2 and RR2 were observed between Day 0 and 7, and between Day 7 and 14, with the greatest proportional changes in RR2. No significant differences in R2* were found., Conclusion: These initial results demonstrate that significant differences in RR2 are detectable after a single week of changes in iron-chelating therapy, likely as a result of superior sensitivity to soluble ferritin iron, which is in close equilibrium with the chelatable cytosolic iron pool. RR2 measurement may provide a new means of monitoring the short-term effectiveness of iron-chelating agents in patients with myocardial iron overload., (Copyright © 2011 Wiley-Liss, Inc.)

Objective: A previous phase 3 study showed significant improvement in walking ability in multiple sclerosis (MS) patients treated with oral, extended-release dalfampridine (4-aminopyridine) 10mg twice daily. The current study was designed to confirm efficacy and further define safety and pharmacodynamics., Methods: This was a 39-center, double-blind trial in patients with definite MS of any course type. Participants were randomized to 9 weeks of treatment with dalfampridine (10mg twice daily; n = 120) or placebo (n = 119). Response was defined as consistent improvement on the Timed 25-Foot Walk, with percentage of timed walk responders (TWRs) in each treatment group as the primary outcome. The last on-treatment visit provided data from 8 to 12 hours postdose, to examine maintenance of effect., Results: One patient from each group was excluded from the modified Intention to Treat population. The proportion of TWRs was higher in the dalfampridine group (51/119 or 42.9%) compared to the placebo group (11/118 or 9.3%, p < 0.0001). The average improvement in walking speed among dalfampridine-treated TWRs during the 8-week efficacy evaluation period was 24.7% from baseline (95% confidence interval, 21.0-28.4%); the mean improvement at the last on-treatment visit was 25.7%, showing maintenance of effect over the interdosing period. There were no new safety findings., Interpretation: This interventional study provides class 1 evidence that dalfampridine extended-release tablets produce clinically meaningful improvement in walking ability in a subset of people with MS, with the effect maintained between doses.

Purpose: To investigate the feasibility of measuring myocardial T2 at 3 Tesla for assessment of tissue iron in thalassemia major and other iron overloaded patients., Materials and Methods: A single-breathhold electrocardiogram-triggered black-blood multi-echo spin-echo (MESE) sequence with a turbo factor of 2 was implemented at 3 Tesla (T). Myocardial and liver T2 values were measured with three repeated breathholds in 8 normal subjects and 24 patients. Their values, together with the T2 values measured using a breathhold multi-echo gradient-echo sequence, were compared with those at 1.5T in the same patients., Results: At 3T, myocardial T2 was found to be 39.6 +/- 7.4 ms in normal subjects. In patients, it ranged from 12.9 to 50.1 ms. "T2 and T2(*) [corrected] were observed to correlate in heart (rho = 0.93, P [corrected] < 0.0001) and liver (rho = 0.95, P < 0.0001). Myocardial T2 and T2 at 3T were also highly correlated with the 1.5T measurements. Preliminary results indicated that myocardial T2 quantitation was relatively insensitive to B1 variation, and reproducible with 3.2% intra-exam and 3.8% inter-exam variations., Conclusion: Myocardial T2 quantitation is feasible at 3T. Given the substantially decreased T2 and increased B0 inhomogeneity, the rapid myocardial T2 measurement protocol demonstrated here may present a robust alternative to study cardiac iron overload at 3T., ((c) 2009 Wiley-Liss, Inc.)

Background: Benign prostatic hyperplasia (BPH) is an age-related enlargement of the prostate, characterized by increased proliferation of stromal and epithelial cells. Despite its prevalence, the etiology of BPH is unknown., Methods: The Brown Norway rat is a model for age-dependent, lobe-specific hyperplasia of the prostate. Histological analyses of the dorsal and lateral lobes from aged rats reveal focal areas characterized by increased numbers of luminal epithelial cells, whereas the ventral lobe is unaffected. This study examined differential gene expression by lobe and age in the Brown Norway rat prostate. The objective was to identify genes with different levels of expression in the prostate lobes from 4-month (young) and 24-month (old) animals, and to subsequently link changes in gene expression to mechanisms of prostate aging., Results: The number of age-dependent differentially expressed genes was greatest in the dorsal compared to the ventral and lateral lobes. Minimal redundancy was observed among the differentially expressed genes in the three lobes. Age-related changes in the expression levels of 14 candidate genes in the dorsal, lateral and ventral lobes were confirmed by quantitative RT-PCR. Genes that exhibited age-related differences in their expression were associated with proliferation, oxidative stress, and prostate cancer progression, including topoisomerase II alpha (Topo2a), aurora kinase B (Aurkb), stathmin 1 (Stmn1), and glutathione S-transferase pi. Immunohistochemistry for Topo2a, Aurkb, and Stmn1 confirmed age-related changes in protein localization in the lateral lobe of young and aged prostates., Conclusion: These findings provide clues to the molecular events associated with aging in the prostate., ((c) 2009 Wiley-Liss, Inc.)

Background: Serum androgen concentrations decline with age in male Brown Norway rats and castration induces apoptosis of luminal secretory epithelial cells in the ventral but not in the dorsal and lateral prostate lobes. Clusterin has been described as an androgen-repressed gene and a protein with either anti- or pro-apoptotic actions., Methods: We measured clusterin mRNA and protein levels, the effects of aging and castration on clusterin protein levels and clusterin immunolocalization within the prostatic ductal network in the prostate lobes of young and aged rats., Results: Whereas levels of clusterin mRNA and protein expression measured by RT-PCR and Western blot, respectively, were higher in the ventral and lateral lobes of aged (24 months) compared to young (4 months) rats, no age-dependent differences were observed in the dorsal lobe. Clusterin expression was localized by immunohistochemistry exclusively to the proximal duct segment of young rats, but extended to the distal segment of the ventral and lateral lobes of aged rats. Despite an age-related decrease in serum testosterone concentration, clusterin gene expression was not altered in the dorsal lobe. After castration, levels of clusterin expression increased significantly in the ventral and lateral lobes despite the absence of epithelial cell apoptosis in the latter. In castrated rats, clusterin expression extended throughout the proximal-distal duct regions of the prostate lobes of young and aged rats., Conclusion: Regulation of clusterin expression in the prostate lobes of aging rats appears complex and is neither directly repressed by androgen nor dependent on apoptotic-induced stress.

Purpose: To optimize the homogeneity and efficiency of the B(1) magnetic field of a four-ring birdcage head coil that is double-tuned at the Larmor frequencies of both (31)P and (1)H and optimized to acquire magnetic resonance spectroscopy (MRS) data at 3T for the study of infants., Materials and Methods: We developed a finite difference time domain (FDTD) tool in-house to iteratively compute and seek the range of geometric and electromagnetic parameters of a dual-tuned, four-ring birdcage coil that would produce the desired resonance patterns, optimize homogeneity of the B(1)-field, and maximize efficiency of the coil. To demonstrate the validity of our computational results, we constructed three RF coils: one dual-tuned coil that was based on the calculated optimized parameters and two single-tuned coils that had dimensions similar to those of the dual-tuned coil, but tuned at the Larmor frequencies of both (31)P and (1)H, respectively. We then tested and compared the performances of the dual-tuned coil and single-tuned coils at both of these frequencies., Results: We found that a dual-tuned, four-ring birdcage coil with a diameter of 180 mm, an inner birdcage length of 100-300 mm, and an outer birdcage length of 25-100 mm produces the desired resonance patterns. For the use of this coil with human infants, optimization of the homogeneity of the B(1) field, combined with improved coil efficiency, yielded a dual-tuned birdcage coil with diameter of 180mm, an inner birdcage length of 150 mm, an outer birdcage length of 25 mm, and corresponding inner and outer capacitances of 17.2 pF and 7.6 pF, respectively. The experimental results from a constructed coil having the same parameters with the modeled coil agreed well with the computational results from the modeled coil. This optimized design overcame the deficiencies of existing dual-tuned, four-ring birdcage coils., Conclusion: The homogeneity and efficiency of the B(1) field for (31)P/(1)H dual-tuned, four-ring birdcage coils can be optimized well using our FDTD tool, especially at high static magnetic fields (B(0)).

Background: Spontaneous age-dependent epithelial cell hyperplasia occurs in the lateral and dorsal, but not the ventral, lobes of aging Brown Norway (BN) rats. Diminished antioxidant enzyme activities and increased formation of reactive oxygen species (ROS) promote the pathology of many aging disorders. We investigated the hypothesis that prostatic epithelial cell hyperplasia in the BN rat was related to age-dependent and/or lobe-specific changes in superoxide dismutase (SOD)., Materials and Methods: Using Western blots, immunohistochemistry and enzyme activity assays we determined the levels of protein expression, subcellular localization, and activities, respectively, of the three SOD isoforms, cytoplasmic SOD1, mitochondrial SOD2, and extracellular SOD3 in the ventral, lateral, and dorsal prostate lobes of 4-month-old rats with normal prostate morphology, in 24-month-old rats with lobe-specific hyperplasia and in older 30-month-old rats., Results: We observed little change in SOD activities as a function of age, although expression of SOD3 increased in the prostatic lobes of older rats. SOD2 levels were higher in the lateral lobe of 4- and 24-month-old rats, but declined by 30 months of age to levels in the ventral and dorsal lobes. SOD1 was localized by immunohistochemistry to the nuclei of epithelial cells in all lobes, but the number of immunopositive nuclei increased in the lateral and dorsal lobes of 24-month-old animals. The concentration of zinc was highest in the prostate lobes of 24-month-old animals., Conclusion: Based upon our data, superoxide dismutase is not significantly altered in the rat prostate during aging and thus is unlikely to be an important factor in the evolution of epithelial cell hyperplasia., ((c) 2005 Wiley-Liss, Inc.)

Mutations of the human androgen receptor gene were identified in five subjects from four families with androgen insensitivity syndrome. Individual exons of the androgen receptor gene were amplified by the polymerase chain reaction from genomic DNA and screened for sequence-dependent differences in their melting characteristics by denaturing gradient gel electrophoresis. DNA fragments from exons with altered mobility were sequenced. Four different single nucleotide base substitutions were found within exons 5, 6, and 7 encoding the steroid-binding domain of the androgen receptor. In one subject with ambiguous genitalia, amino acid residue 763 was changed from tyrosine to cysteine (TAC-->TGC; Y763C). Four subjects, including two siblings, had complete androgen insensitivity. In one subject, residue 779 was changed from arginine to tryptophan (CGC-->TGG; R779W), another subject (M807V) had a substitution of valine (GTG) for methionine (ATG) residue at position 807, and the two siblings (R855C) had a mutation in residue 855 changing arginine (CGC) to cysteine (TGC). Binding of the synthetic androgen ligand, methyltrienolone (R1881), by the mutant receptor Y763C was decreased by 54% compared to the normal receptor. Transcriptional activation of a mouse mammary tumor virus-chloramphenicol acetyltransferase (MMTV-CAT) reporter gene by AR mutant Y763C was negligible at 0.1 nM R1881 and only 55% at 10 nM R1881 when compared to the maximal response with the normal AR, as assessed by CAT activity. Mutant M807V retained only 22% of normal R1881 binding and mutant R855C was unable to bind the steroid. In accordance with the steroid binding, transcriptional activation of MMTV-CAT by M807V rose to only 26% of control in the presence of 10 nM R1881, a concentration at which R855C remained functionally inactive. In summary, missense mutations within the exons of the androgen receptor gene encoding the steroid-binding domain of the receptor are common causes of both partial and complete forms of androgen insensitivity syndrome.

Seventeen patients with patellar retinacular ligament injuries were evaluated with magnetic resonance (MR) imaging. MR findings were correlated with clinical, surgical, and arthroscopic findings. The injuries involved the medial retinacular ligament in 16 of 17 patients and the medial and lateral retinacular ligament in one patient. Nine of 17 patients showed MR evidence of a patellar dislocation. In these patients, a triad of findings that included focal impaction injuries involving the lateral femoral condyle, osteochondral injuries of the medial patellar facet, and injuries of the medial retinacular ligament were seen. Seven of the nine patients with patellar dislocation injuries had frank osteochondral injuries. The injured retinaculum had an indistinct, irregular appearance associated with edema and hemorrhage. The patellar dislocations were clinically unsuspected in the initial evaluation of eight of these nine patients. In eight of 17 patients, the mechanism of injury was a patellar tendon tear (n = 1) or valgus hyperextension (n = 7). In the seven patients with hyperextension injuries, three had associated meniscal and cruciate ligament tears. MR imaging can help define patellar retinacular and associated osteochondral injuries, which may be clinically useful information.

Prior work showed that an intact thyroid axis augments survival from hemorrhagic shock (HS); this study assesses the effects of specific thyroid-related hormones on cardiovascular (CV) function during HS. Following thyroidectomy, 32 conditioned male dogs were subjected to HS to a mean arterial pressure (MAP) of 60 mm Hg for 90 min then to 40 mm Hg for 30 min. Postshock (PS), the dogs received thyroid-releasing hormone (TRH; 2 mg/kg), thyroid-stimulating hormone [control (TSH; 10 IU)], T3 (12 micrograms/kg), or T4 (40 micrograms/kg). Thirty minutes following treatment (PTX), they were resuscitated with shed blood and 50 ml/kg saline. CV and hormonal parameters were measured PS, PTX, postresuscitation (PR), and on day 2 (D2). There were no PS differences in CV parameters between groups. Following treatment, T3 significantly increased MAP (59.0 +/- 13 vs, 39.9 +/- 2.2 mm Hg) and cardiac output (CO; 0.92 +/- 0.1 vs. 0.80 +/- 0.1 liter/min; P < 0.05 by ANOVA). TRH treatment significantly improved PTX MAP (62.7 +/- 10 vs. 40.8 +/- 2.1; P < 0.05 by ANOVA). TSH and T4 did not significantly change PTX MAP or CO. There were no significant CV differences in the four groups following resuscitation or on D2. In conclusion, T3 improves MAP and CO during hemorrhagic shock. TRH transiently improved PTX blood pressure. Further study of the mechanism of this beneficial response afforded by T3 administration is warranted.

Shatney and coworkers, in 1984, reported that recent thyroidectomy in dogs improved survival following hemorrhagic shock induced by reservoir bleeding to a mean pressure (MAP) of 60 torr for 60 min. Since thyroidectomy reduces preshock MAP, their control dogs had a relatively greater fall in MAP during hemorrhage. To circumvent this inequity, this study was designed to elucidate the effects of recent thyroidectomy in 20 splenectomized dogs subjected to acute hemorrhagic shock induced by phlebotomy of a predetermined volume of blood independent of resultant MAP. During splenectomy, ten dogs had thyroidectomy with parathyroid preservation and ten dogs had sham neck operation; 12 days later, hemorrhagic shock was induced by the stepwise bleeding of 35 ml/kg body weight over 75 min. Parameters measured at preshock (baseline), postshock (PS), postresuscitation (PR), and day 2 included MAP, cardiac output (CO), wedge pressure (PCWP), T3, T4, TSH, and hematocrit (Hct). Hypothyroidism was confirmed by low T3 and T4. Baseline and PS hemodynamic parameters were similar for both groups; PR data showed a significant rise in CO (6.7 liter/min vs 3.9 liter/min) and a fall in PCWP (13.2 mmHg vs 18.6 mmHg) in the euthyroid dogs compared to the hypothyroid dogs. Increased CO persisted through day 2 in the euthyroid dogs (3.14 liter/min vs. 1.95 liter/min). Four thyroidectomized dogs died during shock compared to one control dog. Contrary to prior data, this study shows that recent thyroidectomy reduces the survival and compensatory response to hemorrhage. The mechanism leading to this impaired response needs to be defined.

We evaluated testicular function in 15 men with the Martin-Bell (fragile-X) mental retardation syndrome. Macro-orchidism was present in all subjects. Their mean serum LH and FSH levels and plasma testosterone and dihydrotestosterone levels were normal. The mean plasma levels of androstenedione, 17-hydroxyprogesterone, and progesterone were slightly elevated above the normal range, whereas the plasma levels of dehydroepiandrosterone and dehydroepiandrosterone-sulfate were normal. The response in the levels of plasma testosterone following a 5 day period of hCG stimulation was normal in 8 subjects and there was no abnormal accumulation of androgen precursors. The level of 5 alpha-reductase activity and androgen receptor binding was normal in genital skin fibroblasts derived from 3 of these patients. The response of gonadotropin secretion to GnRH stimulation was normal in the 8 men who were tested. Therefore, our data are consistent with the hypothesis that testicular enlargement in men with the Martin-Bell syndrome is not mediated by hormonal factors.

An increase in intracellular pH (pHi) following mitogenic stimulation has been reported in a variety of mammalian cells (W. Moolenaar, Annu. Rev. Physiol., 48:363-376, 1986; E. Rozengurt, Science, 234:161-166, 1986). This increase is currently believed to constitute a "permissive" signal in the process of cell activation (A.E. Lagarde and J.M. Pouyssegur, Cancer Biochem. Biophys. 9:1-14, 1986). Since the majority of studies of this phenomenon have been conducted in the nonphysiological milieu of bicarbonate-free solutions, we have undertaken a study of the effects of bicarbonate and CO2 on mitogen-induced intracellular alkalinization in NIH 3T3 cells. Using nuclear magnetic resonance (NMR) spectroscopy and novel 31P NMR pH indicators (2-amino-phosphono-carboxylic acids) we found that mitogen induces an increase in pHi of 0.16 units only in cells bathed in medium containing low concentrations of bicarbonate (less than 1 mM) and not in cells bathed in medium containing physiological levels of bicarbonate (10-30 mM). In addition to abolishing the mitogen-induced alkalinization, bicarbonate stabilizes pHi at 7.25 units as the external pH (pHe) is varied from 7.0 to 7.6. In contrast, in a bicarbonate-free medium pHi increases from 6.9 to 7.3 over the same range of external pHs. At a constant external pH, increasing the bicarbonate/CO2 concentration results in an increase in pHi from 6.9 in bicarbonate-free solution to 7.25 in a bicarbonate-buffered medium. This relationship is hyperbolic with half-maximal effect occurring at a concentration of 0.4 mM bicarbonate at pH 7.05 and 37 degrees C. Our results suggest that the observations of mitogen-induced alkalinization may be due to the use of nonphysiological bicarbonate-free media. Since this increase in pHi is not observed in physiological media where bicarbonate concentrations are usually greater than 20 mM, we conclude that an increase in pHi is not an obligatory or usual part of the cellular response to growth factors in vivo.