Comparison of biological effects of modulated electro-hyperthermia and conventional heat treatment in human lymphoma U937 cells.

Loco-regional hyperthermia treatment has long history in oncology. Modulated electro-hyperthermia (mEHT, trade name: oncothermia) is an emerging curative treatment method in this field due to its highly selective actions. The impedance-matched, capacitive-coupled modulated radiofrequency (RF) current is selectively focused in the malignant cell membrane of the cancer cells. Our objective is studying the cell-death process and comparing the cellular effects of conventional water-bath hyperthermia treatment to mEHT. The U937 human histiocytic lymphoma cell line was used for the experiments. In the case of conventional hyperthermia treatment, cells were immersed in a thermoregulated water bath, whereas in the case of mEHT, the cells were treated using a special RF generator (LabEHY, Oncotherm) and an applicator. The heating dynamics, the maximum temperature reached (42 °C) and the treatment duration (30 min) were exactly the same in both cases. Cell samples were analysed using different flow cytometric methods as well as microarray gene expression assay and western blot analysis was also used to reveal the molecular basis of the induced effects. Definite difference was observed in the biological response to different heat treatments. At 42 °C, only mEHT induced significant apoptotic cell death. The GeneChip analysis revealed a whole cluster of genes, which are highly up-regulated in case of only RF heating, but not in conventional heating. The Fas, c-Jun N-terminal kinases (JNK) and ERK signalling pathway was the dominant factor to induce apoptotic cell death in mEHT, whereas the cell-protective mechanisms dominated in case of conventional heating. This study has clearly shown that conventional hyperthermia and RF mEHT can result in different biological responses at the same temperature. The reason for the difference is the distinct, non-homogenous energy distribution on the cell membrane, which activates cell death-related signalling pathways in mEHT treatment but not in conventional heat treatment.

Post-immigration Changes in Social Capital and Substance Use Among Recent Latino Immigrants in South Florida: Differences by Documentation Status.

Changing social capital among recent Latino immigrants (RLIs) influences substance use post-immigration. This was a longitudinal study of 476 South/Central American RLIs examining social capital and substance use changes pre to post-immigration. Self-reported measures of social capital and substance use were compared between surveys administered within 1 year of immigration and 2 years post-immigration. Post-immigration, social capital, hazardous drinking and illicit drug use decreased. Women were less likely to engage in hazardous drinking [adjusted odds ratio (AOR) .32, p < .001], and less likely to use illicit drugs (AOR .67, p = .01). Documented individuals with higher levels of 'business' social capital had increased odds of illicit drug use (AOR 2.20, p < .05). Undocumented individuals with higher levels of 'friend and others' social capital had decreased risk for hazardous drinking and illicit drug use (AOR .55, p < .01; AOR .56, p < .05). Documentation status moderated the relationship between social capital and substance use. RLIs can be targeted for primary prevention of substance abuse.

Preoperative evaluation of the deep cerebral veins using 3-tesla magnetic resonance imaging.

BACKGROUND: Surgical treatment of deep-seated tumors such as supratentorial intraventricular and thalamic-pineal-tectal region tumors carries a risk of postoperative deficits due to possible damage to deep cerebral veins including the internal cerebral vein. It is often difficult to identify whether the vessel encountered during surgery needs to be preserved or not through the small operative field. Therefore, preoperative evaluation of deep venous structures is important. We evaluated the usefulness of 3-Tesla magnetic resonance imaging (3 T MRI) for this purpose. METHODS: First, the ability to detect deep venous structures was compared with both 3-dimensional computed tomographical angiography (3D-CTA) and 3 T MRI in patients without any damage to deep venous structures. Images of 7 consecutive patients suffering from insulo-opercular gliomas who underwent both imaging modes for the identification of lateral striate arteries were reconstructed for evaluation of the deep cerebral veins. Subsequently, surgery for tumors at the supratentorial intraventricular and thalamic-pineal-tectal regions was prospectively performed with preoperative evaluation of deep venous system only using 3 T MRI. RESULTS: Information on the deep venous systems acquired by 3 T MRI was as useful as that acquired by 3D-CTA. Until today, we have treated 8 cases of supratentorial intraventricular and thalamic-pineal-tectal region tumors with preoperative evaluation of the deep venous system using 3 T MRI without any morbidity. CONCLUSION: Information on the deep venous system obtained with 3 T MRI aids the surgery of supratentorial intraventricular and thalamic-pineal-tectal region tumors. As the required sequences of 3 T MRI are same as those necessary for the neuronavigation system, and 3 T MRI can be achieved without the use of iodine-based contrast agents, 3 T MRI can be an alternative for preoperative evaluation of the deep venous systems.

A case of idiopathic hypereosinophilic syndrome with leptomeningeal dissemination and intraventricular mass lesion: an autopsy report.

A 43-year-old female presented with idiopathic hypereosinophilic syndrome (HES) manifesting as an intraventricular mass lesion and leptomeningeal and cerebral parenchymal infiltration by eosinophils, lymphocytes and macrophages. She had no history of either malignancy or allergic disorder. She complained of hearing disturbance caused by eosinophilic otitis media. Eosinophilia was detected in the peripheral blood. Hearing disturbance and eosinophilia improved with corticosteroid treatment. Six months later, she was admitted with disturbances of consciousness. Magnetic resonance imaging revealed a mass lesion in the right lateral ventricle and leptomeningeal involvement around the brain stem. Her symptoms deteriorated rapidly, and she died of brain stem malfunction. Autopsy demonstrated significant infiltration by eosinophils and lymphocytes into the mass lesion in the ventricle, subarachnoid space, perivascular space and parenchyma of the medulla oblongata. Histological examination of the bone marrow and other organs did not detect any evidence of parasites, malignancies, or systemic disorders in any organ. The final diagnosis was idiopathic HES. The present case shows that leptomeningeal dissemination and infiltration by eosinophils into the cerebral ventricles and brain stem should be considered in the course of idiopathic HES.

Giant cell reparative granuloma of the proximal tibia: a case report.

Giant cell reparative granulomas (GCRGs) are non-neoplastic inflammatory lesions, usually of the jaw or gingiva or small bones of the hands and feet. We report one such case in the right proximal tibia of a 45-year-old man. Radiological studies showed a lytic lesion with marginal sclerosis in the epiphysis and metaphysis. After open biopsy, a preliminary diagnosis of a benign giant cell tumour was made. One month after admission, the lesion was curetted and filled with cancellous bone and hydroxyapatite. Based on the histology of the curetted lesion, the diagnosis was changed to a GCRG. The patient had an uneventful postoperative course, with no evidence of local recurrence and metastasis. He died from gastric cancer 2 years later.

Ossification of the thoracic ligamentum flavum in an achondroplastic patient: a case report.

We report a case of spinal stenosis with ossification of the thoracic ligamentum flavum in a 53-year-old man with achondroplasia. Neurological signs indicated flaccid paralysis below L1, and the patient was unable to walk. Ossification of the ligamentum flavum was observed at T4/5 and T9 to T12, compressing the thecal sac. Laminectomy of T9 to L1 was performed. At one-year follow-up, the patient was able to walk with one elbow crutch.

Acral myxoinflammatory fibroblastic sarcoma of the foot.

We report a rare case of acral myxoinflammatory fibroblastic sarcoma (AMFS) in a 68-year-old woman. Tumor excision of a mass between the 1st and 2nd toe of the left foot was performed after a diagnosis of ganglion in February 2003. Examination of the surgical specimen confirmed AMFS. No recurrence or metastasis occurred during the follow-up period of 4 years. Clinical characteristics such as recurrence rate, metastasis rate and period of metastasis are unclear for AMFS. Long-term clinical follow-up is thus required.

Genetic analysis of high-metastatic clone of RCT sarcoma in mice, and its growth regression in vivo in response to angiogenesis inhibitor TNP-470.

Genetic analysis of a high-metastatic clone of RCT sarcoma (HM-RCT) was the aim of the study. HM-RCT was developed by the lung passage as well as limiting dilution method from the original RCT sarcoma, in which a tumor was spontaneously developed in a C3H/He mouse. HM-RCT expressed enhanced POU domain (class 2, associating factor 1), adenylate cyclase 7, procollagen type III (alpha), A kinase anchor protein 4 and Ehm (expressed on high-metastatic cells) and 11 expressed sequence tags (ESTs). compared with the original clone of RCT. Eighteen specific genes and 14 ESTs were underexpressed in HM-RCT. We investigated the effects of angiogenesis inhibitor TNP-470 on tumor growth and metastasis of this HM-RCT in vivo. In an experimental group, mice received TNP-470 (30 mg/kg) intraperitoneally every other day. After 5 weeks, the growth of the TNP-470-treated tumor was significantly suppressed in vivo, but did not affect the metastasis. The proportion of positive PCNA-stained cells and cellular telomerase activity was significantly low in response to TNP-470.

Curettage and radiotherapy of giant cell tumour of the sacrum: a case report with a 10-year follow-up.

A case report of a 53-year-old woman with giant cell tumour of the sacrum is presented. Initial curettage was performed through a posterior approach and the patient was relieved of pain and discharged. However, 6 months later the patient was readmitted with a tumour that had progressed towards the L5 vertebra. A further curettage followed by adjuvant radiotherapy resulted in successful reduction of the tumour. Ten years after the operation, there was no recurrence or metastasis.

Cystic lymphangiomas of the cauda equina.

BACKGROUND CONTEXT: Lymphangiomas, benign hamartomatous lesions of the lymph tissue, result from a failure of lymph channels to communicate with the venous system or normal lymph channels. Previously, two reports described lymphangiomas arising in the peripheral nerve (ulnar nerve and posterior tibial nerve). PURPOSE: To report a rare case of cystic lymphangiomas of the cauda equina not previously reported. STUDY DESIGN: Case report. PATIENT SAMPLE: A 56-year-old Japanese man. METHODS: Not applicable. RESULT: The patient recovered from most of his symptoms after tumor extirpation. He had no symptoms at a 5.8-year follow-up. CONCLUSIONS: We believe this is the first case report of lymphangiomas originating from the cauda equina, which was effectively treated by surgery.

(201)Tl-SPECT, (1)H-MRS, and MIB-1 labeling index of central neurocytomas: three case reports.

BACKGROUND: The proliferative activity and metabolic features of three central neurocytomas were investigated using the findings of thallium-201 single photon emission computed tomography ((201)Tl-SPECT) and proton magnetic resonance spectroscopy ((1)H-MRS), and the MIB-1 labeling index (MIB-1 LI). METHOD: The early and delayed (201)Tl indices were calculated as the ratio of tumour to normal brain tissue uptake by (201)Tl-SPECT. In vivo single-voxel (1)H-MRS was performed with echo time of 272 msec to evaluate the metabolites including choline (Cho), N-acetyl aspartate (NAA) and creatine/phosphocreatine (Cre). An external standard reference was used to semiquantitate each metabolite. MIB-1 LI was determined in the surgical specimens. FINDINGS: The MIB-1 LI was 0.5%, 1.2%, and 7.5% in an atypical central neurocytoma without intraventricular extension. Significant (201)Tl uptake was observed on delayed images in all three central neurocytomas. (1)H-MRS showed the high Cho peaks relative to the NAA and Cre peak. The signal at 3.55 ppm, which may be due to inositol or glycine, was observed in one central neurocytoma. INTERPRETATION: Both (201)Tl-SPECT and (1)H-MRS did not reflect the proliferative potential of central neurocytomas.

Developmental changes in the liver weight- and body weight-normalized clearance of theophylline, phenytoin and cyclosporine in children.

AIMS: Body weight- (BW) normalized pediatric dosages of metabolically eliminated drugs often exceed the corresponding adult values. We aimed to clarify whether such findings would be attributable either to an augmented hepatic drug-metabolizing activity or to a systematic bias introduced by adopting BW as a size standard of clearance. MATERIALS AND METHODS: We chose 3 model drugs that are metabolized by distinct cytochrome P450 (CYP) isoforms (theophylline, phenytoin and cyclosporine for CYPIA2, CYP2C9/2C19 and CYP3A4, respectively). The MEDLINE database covering 1966 to May 2001, was searched for articles where systemic clearance oftheophylline or oral clearance of cyclosporine and Vmax/ Km of phenytoin were reported with demographic data of individual children. Liver weights (LWs) of children were estimated using the equation constructed based upon the autopsy data in literature, and body surface area (BSA) was calculated using a standard formula. Relationships between age and clearance of the 3 model drugs that were normalized against BW, LW and BSA were examined. The analysis was confined to the data obtained from children older than 1 year due to scarcity of data for infants and neonates. RESULTS: Relevant data were obtained from 24, 46 and 14 children for theophylline, phenytoin and cyclosporine, respectively. The development of LW lags behind that of BW but is almost identical to that of BSA. Thus, children had a greater LW/BW ratio than adults. The BW-normalized clearance of theophylline and Vmax/Km of phenytoin showed significantly (p < 0.01) negative correlations with age (r = -0.43 and -0.50, respectively) during childhood, whereas their LW- or BSA-normalized clearances were independent of age. CONCLUSIONS: While our analyses were made upon limited numbers of subjects and range of age, the results suggest that children appear to have an augmented BW-normalized clearance for drugs of which metabolism is dominated by the CYP1A2, CYP2C9 or CYP3A4 due mainly to a lagged development of BW than that of LW during childhood. BSA would serve as a practical alternative to LW for scaling adult dosage of metabolically eliminated drugs to children.

Progression of ossification of the posterior longitudinal ligament following en bloc cervical laminoplasty.

BACKGROUND: Ossification of the posterior longitudinal ligament often causes compressive myelopathy. Ossification is a progressive disease, and it has been reported that the area of ossification increases after decompressive surgery. However, it is uncertain how the progression of ossification affects the long-term outcome after cervical laminoplasty. This study was performed to clarify the relationship between the progression of ossification of the posterior longitudinal ligament and the clinical results following en bloc cervical laminoplasty. METHODS: Forty-five patients who were followed for more than ten years after laminoplasty participated in this study. Radiographs and tomograms of the cervical spine of each patient were made before and after the operation and at the time of the latest follow-up. The extent of ossification in the longitudinal and sagittal axes was evaluated. Neurological function was graded with use of the Japanese Orthopaedic Association scoring system. The relationship between the progression of ossification and the score-based rate of recovery was analyzed. RESULTS: Thirty-three (73%) of the patients had progression of ossification of the posterior longitudinal ligament after laminoplasty. Progression was frequent in patients with the mixed type of ossification and in those with the continuous type, whereas it was rare in patients with the segmental type. The patients with progression of the ossification were significantly younger than those without progression (p = 0.018). The Japanese Orthopaedic Association score improved rapidly within one year and continued to improve up to five years after surgery. The score tended to decrease thereafter. For thirteen patients, the score had worsened at the time of the latest follow-up. Three patients had neurological deterioration following an increase in the thickness of the ossification. CONCLUSIONS: Progression of ossification of the posterior longitudinal ligament was often observed during the long-term follow-up period after laminoplasty. Young patients with mixed and continuous types of ossification had the greatest risk for progression. Preventive measures, such as the use of a wider laminar opening during the laminoplasty, should be considered for patients who are at risk for progression of ossification.

Spontaneous reduction in Epstein-Barr virus (EBV) DNA copy number in EBV-infected epithelial cell lines.

We found that spontaneous and 12-0-tetradecanoylphorbol-13-acetate-induced Epstein-Barr virus (EBV) reactivation occurred in short-term (ST)-cultured EBV-infected epithelial cell lines GT38 and GT39 after their establishment; however, it diminished in the long-term (LT)-cultured cells passaged for more than 2 years from ST-cultured cells. We hypothesized that the EBV reactivation may be related to the EBV DNA copy number in the cells. A higher level of EBV DNA content was detected in ST-cultured cells than in LT-cultured cells by Southern hybridization using an EBV DNA XhoI probe. Fluorescence in situ hybridization using EBV DNA BamHI W fragments showed that ST-cultured cells contained a higher EBV DNA copy number than that of LT-cultured cells. EBV DNA-negative cells were detected in small proportions in LT-cultured cells, but were undetected in ST-cultured cells. These results demonstrate that EBV genomes are not maintained stably in the cell lines, and some of them are lost in continuous passages of the cells. We discuss the mechanisms of reduction of EBV reactivation and EBV DNA in the cell lines.

[Increasing fluoroquinolone low-sensitivity in enterotoxigenic Escherichia coli isolated from diarrhea of overseas travelers in Tokyo].

Drug resistance trends were investigated for 1,318 enterotoxigenic Escherichia coli (ETEC) isolated from overseas traveler's diarrheal cases in Tokyo during 1988-1999. A total of 1.6% (21 strains) were nalidixic-acid resistant and fluoroquinolones (NFLX, OFLX, CPFX, LVFX, TFLX, SPFX; FQ) low-sensitive (or low-level-resistant). None of the strains were high-level-resistant to FQ. The FQ low-sensitive strains were isolated in 1996 for the first time, and increased from 3.4% in 1996 to 15.8% in 1999. Countries visited by travelers with the FQ low-sensitive ETEC were India (16 cases), Nepal (3 cases), Cambodia (1 case), and Egypt (1 case). Drug resistance-patterns of the FQ low-sensitive strains, including other drugs (CP, TC, SM, KM, ABPC, ST, NA, and FOM) tested, varied among the 6 types. Among those, multidrug resistant strains accounted for 57.1% (12 strains). The enterotoxin producing types of strains were LT (4 strains), ST (10 strains), and both (7 strains). The serotypes of the strains were classified into 16 types. The quinolone resistance determining regions (QRDRs) of the gyrA genes of the FQ low-sensitive strains were sequenced. The mutations of a Ser to a Leu at position 83 (Ser-83-->Leu) was found in 19 strains, and Asp-87-->Tyr was found in 2 strains.

[Evaluation of animal-assisted therapy for the elderly with senile dementia in a day care program].

We conducted animal-assisted therapy (AAT) for the elderly with senile dementia in a day care center. AAT was performed between July 27 and October 12, 1999 for a total of six biweekly sessions. The AAT group consisted of 7 subjects; 5 with senile dementia of Alzheimer's type (SDAT) and 2 with vascular dementia (VD). The control group was 20 elderly subjects (7 SDAT, 13 VD). The results were as follows: Comparing between MMS scores at the baseline and those three months later, the average score on MMS before AAT (baseline) was 11.43 (+/- 9.00), and that three months later was 12.29 (+/- 9.69). In the AAT group, the average baseline N-ADL score was 28.43 (+/- 14.00) and that after ATT was 29.57 (+/- 14.47). In the AAT group, the average baseline score on Behave-AD was 11.14 (+/- 4.85), and that three months after AAT was 7.29 (+/- 7.11) (p < 0.05). In the control group, the average baseline score was 5.45 (+/- 3.27) and that three month later was 5.63 (+/- 3.59). However, the results of eight subscales of communication behavior three months later were significantly low comparing to those at the baseline in the control group. The evaluation of CgA, which was a mental stress index, showed a decreasing tendency in the AAT group. Our findings suggested we should use several evaluation methods for evaluation of the changes of patients receiving AAT.

Clinical symptoms and surgical outcome in lumbar spinal stenosis patients with neuropathic bladder.

We evaluated the clinical and radiologic features of patients with lumbar spinal stenosis with neuropathic bladder. Based on cystometrogram analysis, the patients were divided into two groups--the neuropathic bladder (NB) group: the NB+ group (23 patients), and the nonneuropathic bladder group: the NB- group (14 patients). The symptom of incontinence was characteristic in patients in the NB+ group. Patients in the NB+ group had a more severe neurologic disturbance, compared with those in the NB- group. The more severe neurologic disturbance was caused by the more striking finding of degenerative spinal stenosis associated with developmental narrowing of the spinal canal. Decompressive surgery had a beneficial effect on the recovery of the neurologic symptoms in both groups. Residual urine volume was reduced after surgery. Postoperative cystometrogram was carried out in nine patients in the NB+ group. It showed a normal pattern in six patients; however, three patients remained in an underactive pattern. Furthermore, four patients still required clear intermittent self-catheterization after surgery.

The fate of autogenous free-fat grafts after posterior lumbar surgery: part 1. A postoperative serial magnetic resonance imaging study.

STUDY DESIGN: Time-related changes in free-grafted fat were investigated by postoperative serial magnetic resonance imaging in 22 patients with degenerative spinal disease who underwent posterior lumbar decompressive surgery. OBJECTIVES: To analyze the size and quality of the grafted fat on magnetic resonance imaging after posterior lumbar surgery. SUMMARY OF BACKGROUND DATA: Epidural fat grafts have been used to prevent epidural and perineural fibroses. Evaluations of changes in grafted fat have been reported, as observed on computed tomography scans. However, there are no published reports of clinical serial magnetic resonance imaging observations of grafted fat after posterior lumbar surgery. METHODS: Axial T1- and T2-weighted magnetic resonance imaging was performed at 3, 7, 21, and 42 days as well as 1 year after surgery. The signal intensity of the fat was classified on the T1-weighted image as Grade 1 (almost equal to subcutaneous fat tissue), Grade 2 (low-signal intensity as compared with subcutaneous fat tissue), Grade 3 (speckled intensity), or Grade 4 (signal void, suggesting the change to scar tissue. The time-related, cross-sectional area of the subarachnoidal space was measured from the T2-weighted image. RESULTS: The size of the grafted fat was reduced to approximately 57% after 42 days, and to about 33% after more than 1 year, as compared with the condition 3 days after surgery. In time, the shape of the grafted fat was changed along the shape of the dura mater. During the early stage (within 6 weeks after surgery), the signal intensity of the grafted fat was lower than that of normal subcutaneous fat tissue (Grades 2-4: 40.9-59.1%). However, the intensity had recovered to normal status (Grade 1) by 1 year after surgery. CONCLUSIONS: The total amount of grafted fat used in posterior lumbar surgery is reduced. However, as observed on magnetic resonance imaging, it is alive and remodeled along the shape of the dura mater in relation to its shrinkage and reexpansion. This remodeling of the grafted fat is meaningful and effective in protecting the spinal nerve. This report clarifies the fate of the grafted fat from the findings of postoperative serial magnetic resonance imaging after lumbar decompressive surgery.

The fate of autogenous free-fat grafts after posterior lumbar surgery: part 2. Magnetic resonance imaging and histologic studies in repeated surgery cases.

STUDY DESIGN: Histologic changes of free-grafted fat were investigated from surgical specimens taken at repeated lumbar surgery in 18 patients with degenerative spinal disease who previously had undergone posterior lumbar decompression and free fat graft. OBJECTIVE: To clarify the clinical usefulness of free-grafted fat by histologic analysis. SUMMARY OF BACKGROUND DATA: The clinical postoperative serial magnetic resonance imaging studies in Part 1 showed that the size of grafted fat was reduced, and that the shape changed along with the shape of the dura mater. The shape of the grafted fat was remodeled in relation to the postoperative transient shrinkage and reexpansion of the dura mater. METHODS: From repeated surgery cases, the status of the previously grafted fat tissue in the virgin operation was analyzed. Sections from the specimens resected surgically during repeated surgery were stained with hematoxylin and eosin or osmium. The size and quality of the grafted fat globules were analyzed by computer. RESULTS: In all cases, grafted fat tissue was identified as a survival. Hematoxylin and eosin staining showed increased collagen fiber and hyperplasia of blood vessels entering the fat tissue that survived. From osmium staining, the size of fat globules was reduced. The shape and quality of the fat globules were analyzed by original staging. They showed various sizes, polymorphism, and vacuolar degeneration. CONCLUSIONS: Finding showed that the grafted fat used in posterior lumbar surgery reduced the size and quality of the fat globules, as compared with normal fat tissue. However, the grafted fat tissues were confirmed to be alive over the long term. This study clarified the fate of the grafted fat as observed by magnetic resonance imaging and histology from repeated surgery cases.

A day care program and evaluation of animal-assisted therapy (AAT) for the elderly with senile dementia.

We conducted a survey to clarify the evaluation methods of animal-assisted therapy (AAT) for the elderly with senile dementia in an adult day care center. AAT was implemented for a total of six biweekly sessions. The AAT group consisted of seven subjects and the control group numbered 20 subjects. In a comparison between Mini-Mental State Exam (MMSE) scores at baseline and those measured three months later, the average MMSE score before AAT (baseline) was 11.43 (+/- 9.00), and three months later it was 12.29(+/- 9.69). In the AAT group, the average score on Nishimura's Activities of Daily Living (N-ADL) at baseline was 28.43(+/- 14.00), and after ATT it was 29.57(+/- 14.47). In the AAT group, the average baseline score on behavioral pathology of Alzheimer's disease (Behave-AD) was 11.14(+/- 4.85), and three months after AAT it was 7.29(+/- 7.11) (p < 0.05). In the control group, the average baseline score was 5.45(+/- 3.27) and three months later it was 5.63(+/- 3.59). The evaluation of salivary CgA, as a mental stress index, showed a decreasing tendency in the AAT group. Our findings demonstrate the usefulness of using several methods for evaluation of the changes in patients given AAT.