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Partial inversion of the initiation-promotion sequence of multistage tumorigenesis in the skin of NMRI mice.

Alterations in NMRI mouse skin induced by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate in "stage I of tumor promotion" are slowly reversible, and this reversibility has a half-time of 10 to 12 weeks. The tumor response observed in the course of an initiation-promotion experiment in vivo is independent of whether stage I of promotion occurs before or after initiation. Since the time interval between treatment with the promoter, and subsequent initiation can be extended up to at least 6 weeks, an enhancement of initiation because of promoter-induced cellular DNA synthesis seems to be unlikely. This result may be inconsistent with the two-stage model of tumor promotion because it indicates that in skin the existence of initiated cells is not required for the induction of cellular alterations that are essential for the stage of skin tumorigenesis called stage I of promotion.

9,10-Dimethyl-1,2-benzanthracene↗

Isolation, characterization, and analysis of the expression of the cbhII gene of Phanerochaete chrysosporium.

Two cDNA sequences representing putative allelic variants of the Phanerochaete chrysosporium cbhII gene were isolated by hybridization to the Trichoderma reesei cbhII gene. Both of the equivalent genomic sequences were subsequently isolated by the inverse PCR technique. DNA sequencing showed that the cbhII open reading frame of 1,380 bp codes for a putative polypeptide of 460 amino acids which is interrupted by six introns. The domain structure found in T. reesei cbhII is conserved in the equivalent P. chrysosporium protein. The overall similarity between the two gene products is 54%, with the region of highest conservation being found in the cellulose-binding domain (65%). Unlike the cbhI gene of P. chrysosporium, cbhII does not appear to be a member of a class of closely related genes. CBHII is a new member of family B of the beta-1, 4-glucanases. Alignment of the P. chrysosporium and T. reesei CBHII protein sequences showed that all of the residues important for the formation of the extended loops of the catalytic domain and those residues that are involved in the catalytic action of the T. reesei enzyme are also present in the P. chrysosporium equivalent. The profiles of cbh gene expression in P. chrysosporium reveal that while cbhI.1 and cbhI.2 could be coregulated, cbhII can be independently controlled. The latter is so far the only cellulase gene found to be expressed when the fungus is grown on oat spelt arabinoxylan, suggesting that it may play an active role in the xylanolytic as well as the cellulolytic systems.

Amino Acid Sequence↗

A strategy for magnetic resonance imaging of the head: results of a semi-empirical model. Part II.

A semi-empirical model was used to identify specific pulse sequences that cause most lesions to appear distinctly brighter than normal tissues in magnetic resonance (MR) images of the head. Clinical trials confirm the utility of these sequences for patient screening. As a result, a strategy for effective and efficient MR imaging of the head is proposed. The previously described gray-scale model has been modified to account for the effect of image noise. By means of computer simulation, 13,800 different hypothetical cerebral lesions were imaged with a variety of pulse sequences. A number of conclusions resulted. First, two sequences are expected to be sufficient to visualize most intracranial lesions, a "diagonal" SE sequence (e.g., SE 2500/80) and an IR sequence with a short inversion time (e.g., IR 1800/200). These sequences are orthogonal, i.e., lesions missed by one are likely to be detected by the other. Second, signal averaging the screening sequences is expected to be more effective than optimized sequences when lesion tissue parameters differ little from brain. Finally, the effectiveness of unaveraged screening sequences suggests that improved signal-to-noise ratio (SNR) is not necessary for the detection of most large lesions. Therefore, the increased SNR achievable through signal averaging or increased field strength might best be utilized to improve spatial resolution so that smaller lesions can be detected.

Brain Diseases↗

Liver metastases: optimization of MR imaging pulse sequences at 1.0 T.

Twenty patients with known liver metastases were examined with magnetic resonance imaging; four different pulse sequences were used and six different images were produced to allow comparison of pulse sequence performance at a 1.0-T field strength. Pulse sequence performance was in each case calculated by measuring contrast-to-noise ratios (C/N) comparing normal liver, metastatic tumors to liver, and background noise. All pulse sequences required approximately the same length of time for data acquisition. Short inversion time inversion-recovery (STIR) sequences yielded the greatest signal difference-to-noise ratio in 17 patients. Phase contrast images produced the greatest contrast ratio in three patients. Although short TR, short TE spin-echo sequences provided the best anatomic detail, in no patient did this sequence yield the greatest signal difference-to-noise ratio in the comparison of liver and tumor. Differences in field strength and in equipment software and hardware may account for the discrepancy between our findings and previously published data.

Humans↗

Reduction of CSF artifacts on FLAIR images by using adiabatic inversion pulses.

The purpose of this study was to investigate the possibility that some artifactual high signals produced in CSF with fluid-attenuated inversion-recovery MR sequences could be due to inhomogeneity in the amplitude of the initial inversion pulse, and that this problem could be reduced or eliminated by the use of adiabatic inversion pulses. Studies with four volunteers showed dependence of high CSF signals in the posterior fossa on radiofrequency pulse amplitudes and that these signals could be eliminated by the use of adiabatic inversion pulses. Two illustrative clinical cases are included.

Adult↗

Low-level inversion of the L component of pseudorabies virus is not dependent on sequence homology.

Pseudorabies virus has a class 2 genome in which the S component is found in two orientations relative to the L component. The L component is bracketed by sequences that are partially homologous; it is found mainly in one orientation, but a small proportion is inverted (J. M. DeMarchi, Z. Lu, G. Rall, S. Kuperschmidt, and T. Ben-Porat, J. Virol. 64:4968-4977, 1990). We have ascertained the role of the patchy homologous sequences bracketing the L component in its inversion. A viral mutant, vYa, from which the sequences at the right end of the L component were deleted was constructed. Despite the absence of homologous sequences bracketing the L component in vYa, its L component inverted to an extent similar to that of the L component in the wild-type virus. These results show the following. (i) The low-frequency inversion of the L component of PrV is not mediated by homologous sequences bracketing this component. (ii) Cleavage of concatemeric DNA at the internal junction between the S and L components is responsible for the appearance of the minority of genomes with an inverted L component in populations of pseudorabies virus. (iii) The signals present near or at the end of the S component are sufficient to allow low-frequency cleavage of concatemeric DNA; the sequences at the end of the L component are not essential for cleavage, although they enhance it considerably.

Animals↗

Herpesvirus-dependent amplification and inversion of cell-associated viral thymidine kinase gene flanked by viral a sequences and linked to an origin of viral DNA replication.

The genome of herpes simplex virus 1 or 2 consists of two components, L and S, which invert relative to each other during infection. As a result, viral DNA consists of four equimolar populations of molecules differing solely in the relative orientations of the L and S components. Previous studies have shown that the a sequences, located in the same orientation at the genomic termini and in inverted orientation at the L-S junction, play a key role in the inversion of L and S components. In this report we describe a virus-dependent system designed to allow identification of the viral genes capable of acting in trans to invert DNA flanked by inverted copies of a sequences. In this system, cells are converted to the thymidine kinase-positive phenotype with a chimeric plasmid carrying the thymidine kinase gene flanked by inverted copies of the a sequence and linked to an origin of viral DNA replication derived from the S component. The DNA introduced into the cells is retained and propagated in its original sequence arrangement as head-to-tail concatemers. Infection of these cells with herpes simplex virus 1 or 2 results in as much as 100-fold amplification of the plasmid sequences and inversion of the DNA flanked by copies of the a sequence. In infected cells, the amplified resident DNA accumulates in head-to-tail concatemers and no rearrangement other than the inversions could be detected. These results suggest that the a sequence-dependent inversions required trans-acting viral gene products.

Animals↗

Evaluation of intracranial lesions with inversion recovery half-Fourier single-shot turbo spin-echo MR: initial observations.

PURPOSE: To determine the value of inversion recovery half-Fourier single-shot turbo spin-echo (IR-HASTE) MR sequences in the characterization of a variety of intracranial lesions, with the focus on differentiating between epidermoids and nonneoplastic cystic lesions. METHODS: We used a 1.5-T MR unit to study five epidermoids, seven arachnoid cysts, seven other nonneoplastic cysts (three neuroepithelial cysts, two interhemispheric cysts, and two Rathke's cleft cysts), and eight solid neoplasms (three meningiomas, two astrocytomas, one subependymoma, one cavernoma, and one metastatic tumor) using IR-HASTE sequences with variable inversion times (TI). Imaging time for each section was 2 seconds for the sequence. The TI nulling values were analyzed statistically. RESULTS: The TI nulling values were 1200 to 2300 for the epidermoids, 2800 to 3000 for the arachnoid cysts, 300 and 800, respectively, for the Rathke's cleft cysts, 2500 to 3000 for the other nonneoplastic cysts, and 300 to 1500 for the solid neoplasms. There was no overlap of TI nulling values between the arachnoid cysts and the epidermoids; the difference was statistically significant. Both patients with interhemispheric cysts had two lesions in which the TI nulling values were different. CONCLUSION: IR-HASTE sequences provide a rapid and reliable imaging method for differentiating among epidermoids, arachnoid cysts, and solid neoplasms. This technique also provides information about the continuity of the multicystic lesions in terms of the differences in their TI nulling values. For solid intraaxial masses, the use of IR-HASTE helps to differentiate intratumoral cysts and necrosis from solid components.

Adolescent↗

TurboFLASH FAIR imaging with optimized inversion and imaging profiles.

Optimal implementation of pulsed arterial spin labeling (PASL) methods such as flow-sensitive alternating inversion recovery (FAIR), require the minimization of interactions between the inversion and imaging slabs. For FAIR, the inversion:imaging slice thickness ratio (STR) is usually at least 3:1 in order to fully contain the extent of the imaging slice. The resulting gap exacerbates the transit time. So far, efforts to minimize the STR have concentrated on the inversion profile. However, the imaging profile remains a limiting factor especially for rapid sequences such as turbo fast low-angle shot (TurboFLASH) which uses short pulses. This study reports the implementation of a TurboFLASH sequence with optimized inversion and imaging profiles. Slice-selection is achieved with a preparation module incorporating a pair of identical adiabatic frequency offset corrected inversion (FOCI) pulses. The optimum radiofrequency (RF) and gradient scheme for this pulse combination is described, and the relaxation characteristics of the slice-selection scheme are investigated. Phantom experiments demonstrate a reduction in the STR to approximately 1.13:1. Implementation in an animal model is described, and the benefit of the improved profile in probing the sensitivity of the flow signal to tagging geometry is demonstrated. Sensitivity to transit time effects can be minimized with this sequence, and ASL methodologies can be better explored as a result of the improved conformance with the ideal of square slice profiles.

Animals↗

In vivo tumor lactate relaxation measurements by selective multiple-quantum-coherence (Sel-MQC) transfer.

The frequency-selective multiple-quantum-coherence (Sel-MQC) lactate (Lac) filter offers complete lipid and water suppression in a single scan for robust in vivo detection of tumor Lac, even in the presence of abundant lipids. Conversion of the detected signal into accurate tissue concentrations of Lac requires knowledge of in vivo Lac T1 and T2 relaxation times. This work reports modifications to the Sel-MQC pulse sequence, T1- and T2-Sel-MQC, that facilitate relaxation measurements of Lac. The T1-Sel-MQC sequence combines an inversion prepulse with the Sel-MQC filter. The T2-Sel-MQC sequence incorporates a CH3-selective 180 degrees pulse during the MQ preparation period to overcome the J-modulation effects and allow the insertion of variable echo delays. The performance of these sequences was evaluated with the use of phantoms and subcutaneous murine tumor models in vivo. The present approach will allow investigators to correct for the relaxation-induced Lac signal loss in Sel-MQC experiments for the quantitative mapping of in vivo tumor Lac distribution.

Animals↗

The value of new MRI techniques in adrenoleukodystrophy.

PURPOSE: To evaluate the significance of new techniques in MRI examinations of the brain in children with X-linked adrenoleukodystrophy (ALD). MATERIALS AND METHODS: Five patients aged between 2 and 15 years with childhood ALD were examined with MRI. Following conventional spin-echo (CSE) sequence, a fast spin-echo (FSE) sequence with T2 contrast was performed in four patients, an FSE turbo-inversion-recovery (TIR) sequence in two, magnetization transfer contrast (MTC) imaging in two, and localized proton spectroscopy (MRS) in four patients. RESULTS: FSE compared favorably with CSE with a time saving of 70%. On TIR images the contrast between normal and demyelinated white matter was greater than on CSE and FSE images. Calculated MTC values revealed a severe MTC loss within the demyelinated regions and a moderate reduction in the border zones. In this way, calculation of MTC might be useful to differentiate between edematous changes and areas of irreversible demyelination. MRS revealed a reduction in N-acetylaspartate and an elevation in choline (Cho). The degree of MRS changes paralleled the severity of demyelination. A Cho elevation may precede visible demyelination on T2-weighted images. CONCLUSIONS: T2-weighted FSE sequences can replace CSE without any disadvantages and with effective time saving. The indication for MTC imaging and MRS in children with ALD is not yet finally defined. These new techniques may reveal the earliest signs of cerebral involvement or of disease progression, a matter of great importance in selecting the optimal therapeutic approach.

Adolescent↗

[MRI of osteomyelitis: sensitivity and specificity of STIR sequences in comparison with contrast-enhaned T1 spin echo sequences].

PURPOSE: To evaluate the need for additional MR sequences including administration of Gd-DTPA after inconspicuous Short-Tau Inversion-Recovery (STIR) sequence to exclude the diagnosis of osteomyelitis. MATERIAL AND METHODS: 112 MR examinations of 79 patients acquired for the detection of possible osteomyelitis were analyzed retrospectively. All examinations were performed at 0.5 T including STIR, T1-weighted spin echo sequences (T1 SE) before and after application of Gd-DTPA. Additionally, 93 T2-weighted spin echo sequences were available. The examinations were analyzed by two experienced radiologists. First, the STIR sequences were studied, followed by the T1 SE images before and after administration of contrast material. Finally, the T2-weighted images were evaluated. Diagnoses were confirmed by operation (22), biopsy (10), and follow-up (80). RESULTS: In 53 cases osteomyelitis was diagnosed, while the remaining 59 cases suffered from another disease. The sensitivity of the STIR sequence was 100% while the specificity for osteomyelitis was 49.2%. The specificity increased to 79.7% by including T1 SE images into the analysis and reached 83.1% after considering the contrast enhanced images. T2-weighted images yielded no additional information. CONCLUSION: The combination of STIR and T1SE images shows a high sensitivity and specificity for osteomyelitis, thus obviating the need for any additional examinations.

Adolescent↗

Asymptomatic spinal cord lesions in clinically isolated optic nerve, brain stem, and spinal cord syndromes suggestive of demyelination.

OBJECTIVES: Conventional T2 weighted MRI studies have highlighted the fact that the presence of clinically silent brain lesions increases the risk of developing clinically definite multiple sclerosis after an isolated syndrome of the optic nerve, brain stem, or spinal cord. The objectives of the present study are: (1) to show whether or not these patients also have asymptomatic abnormalities of the spinal cord, and (2) to recruit a new cohort of such patients using high resolution MRI of both brain and spinal cord. METHODS: The brain was imaged in the axial plane with 3 mm thick contiguous slices using a proton density and T2 weighted fast spin echo (FSE) sequence; a T1 weighted sequence after the injection of gadolinium-DTPA; and a fast fluid attenuated inversion recovery (fFLAIR) sequence. The spinal cord was imaged in the sagittal plane with 3 mm thick slices using a T2 weighted FSE and a T1 weighted gadolinium enhanced sequence. RESULTS: Thirty three patients, mean age 31 (16-46) were recruited. There were 14 men and 19 women. Brain MRI was abnormal in 22 (67%); no patient was seen with abnormalities on only one or other sequence. Six patients (18%) displayed one or more gadolinium enhancing lesions on brain MRI. In the spinal cord, nine (27%) patients displayed one or more clinically silent lesions on FSE. Two patients showed one and two gadolinium enhancing lesions in the spinal cord respectively. CONCLUSION: This high incidence of spinal cord lesions emphasises that asymptomatic demyelinating lesions may also involve clinically eloquent pathways. Follow up studies are required to determine their prognostic importance.

Adolescent↗

Application of spin-spin relaxation to measurement of surface area and pore size distributions in a hydrating cement paste.

Nuclear magnetic resonance (NMR) relaxation analysis has been applied to interpret the evolution of microstructure in a cement paste during hydration. Measurements of transverse magnetic relaxation were made in fully and partially filled white cement pastes for hydration times between 1 h and 6 mo. It was found that only the evaporable water molecules contributed to the echo amplitude in the NMR measurement, while both the evaporable and the hydrated water contribute to the amplitude of the free induction decay. The spin-spin relaxation rate was found to increase markedly during hydration and can be directly related to the total surface area of the CSH gel accessible to evaporable water. The amount of water consumed during the hydration process was determined independently from the amplitude of the NMR echo signal extrapolated to the beginning of the pulse sequence. A numerical inversion of the full spin-spin relaxation profile, determined with a CPMG pulse sequence, was interpreted as a pore volume distribution function. The measurements of total surface area, amount of water consumed, and pore distributions were performed in situ nondestructively, throughout the hydration period for a single sample of cement paste.

Construction Materials↗

Centromeric alphoid sequences are breakage prone resulting in pericentromeric inversion heteromorphism of qh region of chromosome 1.

Structural variations in the pericentromeric region of chromosome 1 are considered the norm. We characterized a chromosome 1 with an inversion by FISH-technique and suggested that the origin of pericentromeric heteromorphism is far more complex than previously suggested. It is postulated that similar to the centromeric alphoid DNA sequences of chromosome 9, chromosome 1 also possesses a "breakage prone" centromeric domain which may be a possible cause of such inversions involving the secondary constriction region.

Adult↗