In vitro and in vivo activity of murine lymphokine-activated killer cells after cryopreservation

The in vitro and in vivo effects of cryopreservation on the cytotoxic activity of murine lymphokine-activated killer (LAK) cells were studied. LAK cells were generated by incubation of spleen lymphocytes of BALB/c mice for 3 days with recombinant interleukin-2 (rIL-2) and subsequent cryopreservation. Cytotoxicity was determined in a 51Cr release assay. After thawing, cytotoxic activity was reduced (40.4% 51Cr release at an effector:target cell ratio of 40:1 as compared to 68.5% 51Cr release before freezing) and could be restored to precryopreserved levels by reincubation with rIL-2 for 2 days after thawing (78.8% 51Cr release). These cells were then tested in BALB/c mice injected with RAW 112 cells, a pre-B-cell lymphoma line. The results demonstrate that the survival rate of mice injected with cryopreserved and restimulated LAK cells (50% survival greater than 180 days after injection) did not differ significantly from that of mice injected with fresh unfrozen LAK cells (60% survival greater than 120 days, 50% survival greater than 180 days). Cryopreserved LAK cells have potential use in adoptive immunotherapy.     

Intrasession reliability of force platform parameters in community-dwelling older adults

Bauer C, Gröger I, Rupprecht R, Gaßmann KG. Intrasession reliability of force platform parameters in community-dwelling older adults.

Objective

To investigate the intrasession reliability of center of pressure (COP) parameters calculated from force platform measurements.

Design

A cross-sectional study.

Setting

Gait and balance laboratory.

Participants

Community-dwelling healthy older adults (N=63) above the age of 62 years (mean age, 78.74y).

Interventions

Not applicable.

Main Outcome Measures

COP was estimated from a force platform, and the following parameters were calculated: (1) the total length of the COP displacement, (2) area of sway, (3) length of the COP displacement in the sagittal plane, and (4) length of the COP displacement in the frontal plane. Intraclass correlation coefficients (ICCs) were calculated by using 3 successive trials with 4 different test conditions. The test conditions were (1) normative standing with eyes open, (2) normative standing with eyes closed, (3) narrow stance with eyes open, and (4) narrow stance with eyes closed.

Results

The ICCs for the tests with eyes closed (.710-.946) were higher than those for tests with eyes open (.841-.945). The highest value was obtained for the vector sum of the COP during anteroposterior movement in narrow stance with eyes closed (.946). The value .710 was the lowest of all parameters and was an outlier for the narrow stance with eyes closed test, which was otherwise very reliable.

Conclusions

Eight of 16 calculated ICCs showed excellent reliability (>.90). They can be recommended for further use in clinical trials. Tests with closed eyes were more reliable than tests with eyes open. We recommend using eyes closed test conditions when assessing static balance control. For these tests, all the calculated ICCs were over .90, except for measurements of sway area.     

One-year course of paediatric headache in children and adolescents aged 8-15 years

The aims of the present study were the assessment of headache (HA) prevalence in German children and adolescents in the second year of a 4-year longitudinal study and the analysis of headache status change from year 1 to year 2. The original sample consisted of 8800 households with a child aged 7-14 years. A total of 4159 households responded in both year 1 and year 2, yielding 3984 valid parent questionnaires. Data regarding various aspects of the child's HA history and general health were gathered via mailed questionnaires from the parents. Of the households returning valid parent questionnaires at survey 2, 48.9% reported their child to have experienced headaches during the previous 6 months (53% at survey 1). Weekly HA was reported for 6.5% of the children, monthly or less frequent HA for 16.5% and 25.9%, respectively. With regard to headache diagnosis, 55.0% of the children and adolescents with HA experienced tension-type HA (TTH) and 11.3% migraine with or without aura (M). For more than half of the children and adolescents with HA (57.0%) the frequency of head pain remained stable over the period of 1 year (i.e. same frequency category in years 1 and 2). Improved and worsened HA status regarding frequency of occurrence was found in 22.3% and 20.7% of the subjects, respectively. Thus, there was no definite trend towards an increase of HA episodes over the course of 1 year regarding the individual child or adolescent. The most stable type of HA was TTH.     

Novel antibodies directed against the human erythropoietin receptor: creating a basis for clinical implementation

Recombinant human erythropoietin (rHuEPO) is an effective treatment for anaemia but concerns that it causes disease progression in cancer patients by activation of EPO receptors (EPOR) in tumour tissue have been controversial and have restricted its clinical use. Initial clinical studies were flawed because they used polyclonal antibodies, later shown to lack specificity for EPOR. Moreover, multiple isoforms of EPOR caused by differential splicing have been reported in cancer cell lines at the mRNA level but investigations of these variants and their potential impact on tumour progression, have been hampered by lack of suitable antibodies. The EpoCan consortium seeks to promote improved pathological testing of EPOR, leading to safer clinical use of rHuEPO, by producing well characterized EPOR antibodies. Using novel genetic and traditional peptide immunization protocols, we have produced mouse and rat monoclonal antibodies, and show that several of these specifically recognize EPOR by Western blot, immunoprecipitation, immunofluorescence, flow cytometry and immunohistochemistry in cell lines and clinical material. Widespread availability of these antibodies should enable the research community to gain a better understanding of the role of EPOR in cancer, and eventually to distinguish patients who can be treated safely by rHuEPO from those at increased risk from treatment.     

Adequate cytogenetic examination in myelodysplastic syndromes: analysis of 529 patients

In myelodysplastic syndromes (MDS), the karyotype is one of the most significant prognostic markers with profound impact on differential diagnosis and therapeutic decisions. In a retrospective study, we examined karyotypes of bone marrow specimens of an oligocentric cohort comprising 529 patients with MDS to address the question how many metaphases need to be analyzed to detect even small cell clones with an appropriate expenditure. We found a statistically significant difference of the frequency of normal karyotypes in the patient group with 19 or less analyzed metaphases compared to the group with 20 or more metaphases analyzed (56% versus 47%, p=0.041). Furthermore, we demonstrate that the analysis of 25 or more metaphases can further improve the sensitivity of karyotype analysis and leads to the identification of additional clinically relevant abnormal clones or subclones in a substantial proportion of patients. In summary, our data suggest the examination of at least 20 metaphases in MDS.     

Value of preoperative diagnostic modalities in patients with recurrent thyroid carcinoma

BACKGROUND: Patients with well-differentiated thyroid cancer (WDTC) regularly have an excellent prognosis. However, tumor recurrence either involving the thyroid bed or the regional lymph nodes, or both, can be associated with significant morbidity and even mortality. The aim of the follow-up after primary surgery is to detect recurrent disease at its earliest stage. We assessed the value of different diagnostic methods in detecting locoregional recurrence in patients with WDTC. METHODS: We prospectively identified 150 patients with WDTC. Of those, 43 (28.7%) presented with recurrent disease. Ultrasonography-guided fine needle biopsy (US-FNB), iodine 131 ((131)I) wholebody scintigraphy, thyroglobulin (Tg) measurement, and fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) were carried out. RESULTS: Ultrasonography detected malignant lesions in 95.3% of the patients. The true positive rate of US-FNB was 95.3%. (131)I scanning had true positive, false negative, and false positive results in 54.2%, 40.0%, and 5.7% of the cases, respectively. In 85.7% of the patients, Tg levels were within pathologic range. Among the 13 patients who underwent FDG-PET, 84.6% showed pathologic uptake indicating malignancy. US and US-FNB provided the highest specificity for detecting recurrence (P <.001). CONCLUSIONS: In patients with WDTC and locoregional recurrence, US and US-FNB are the most sensitive methods in detecting local recurrence or regional lymph node metastases. FDG-PET is valuable in case of negative (131)I scanning results and elevated serum Tg levels. The method has limitations in finding minimal disease.     

Cerebral Tuberculoma

SUMMARY Intracranial tuberculoma has become a rarity. It remains a curable lesion that responds well to medical therapy. Although diagnosis in developed countries is often made only postoperatively, early and effective treatment can be instituted if a high index of suspicion is maintained and diagnostic criteria are looked for. A case is presented which illustrates the difficulties in reaching a diagnosis, and a review of the literature is given.     

Study on the effect of oral administration of carbocisteine on ventilatory parameters in the SO2 inhalation model of bronchitis in the rat

In order to study the physiological correlates of the beneficial action of carbocisteine (S-carboxy-methyl-cysteine), we have measured the changes occurring in ventilatory parameters in rats made bronchitic by prolonged exposure (2 weeks) to air containing sulfur dioxide (SO2). In animals treated with distilled water (1 ml/100 g/day), statistically significant (P < 0.05) changes in respiratory frequency (-20%) and tidal volume (+31%) were found. As a result of these opposing changes, the ventilation/min was stable. Moreover, the compliance was decreased (33%, P < 0.05) and the resistance was greatly enhanced (+ 99%, P < 0.05). The concomitant administration of carbocisteine (500 mg/kg po/day) with SO2 inhalation significantly (P < 0.05) prevented the development of resistance without effecting significant changes in the other parameters except for a slight improvement in ventilation/min. In conclusion, this improved respiratory resistance in the bronchitic carbocisteine-treated animals tallies with a decrease in mucus retention associated with the return to normal of rheological characteristics of the secreted mucus.