Detection and characterization of the electron paramagnetic resonance-silent glutathionyl-5,5-dimethyl-1-pyrroline N-oxide adduct derived from redox cycling of phenoxyl radicals in model systems and HL-60 cells

Rapidly growing knowledge about the nature and behaviour of breast cancer has led to many treatment modalities. Consequently, the possibilities of individualizing the treatment of breast cancer increase. The major tool for the determination of an optimal treatment plan is the estimation of the extent of the disease: in other words, staging. As a consequence, together with the expected result of the treatment, the stage of the disease gives information on the prognosis of the patient. Current staging systems insufficiently describe the clinically important features of breast cancer with respect to management and outcome: local and regional extent, invasiveness, aggressiveness, the state of dissemination, and the effectiveness of different treatment modalities. For staging of the local and regional extent, histology plays a prominent role and should be incorporated in future staging systems. Histological workup therefore needs standardisation. Histological parameters as tumour size, grade, nodal status, and vascular invasion are also the most important prognostic factors. Many so-called biological prognostic factors are related to the invasiveness and aggressiveness (metastatic potential) of the tumour, and therefore to the prognosis of the patient. However, these factors do not necessarily predict the effectiveness of certain systemic treatments. Only if the biological foundation of a prognostic factor is completely clarified can treatment be based on this knowledge, and the factor will become a predictor for the treatment effect. Many "biological" prognostic factors do not fulfil this main criterion and are therefore not useful for clinical decision making. A clinically useful staging system covers three primary aims: (1) to guide locoregional treatment, (2) to prognosticate the chance of survival, and (3) to indicate who needs what kind of adjuvant treatment. For the conception of a new staging system the following steps should be taken: standardization of all aspects of histology, identification of regional nodal involvement, and validation of prognostic factors with respect to their predictive value to treatment outcome.     

Utilizing uncertainty factors in minimal risk levels derivation

While both 31P and 113Cd are present at locations of interest in many different macromolecular systems, heteronuclear-detected relaxation measurements on these nuclei have been restrained by limitations in either resolution or signal-to-noise ratio. We have developed heteroTOCSY-based methods to overcome both of these problems. Two-dimensional versions of these experiments were utilized to measure 31P T1 and T2 values in DNA oligonucleotides; the additional resolution offered by a second dimension allowed determination of these values for most of the 31P resonances in a DNA dodecamer. The results from the experiments indicated that there was little significant variation in T1 values for the different phosphates in the DNA dodecamer; however, the T2 values showed a clear pattern, with lower values in the interior of the sequence than at the ends of the helix. Furthermore, a significant correlation between 31P chemical shifts and T2 values was observed. One-dimensional, frequency-selective versions of these experiments were also developed for use on systems containing a smaller number of heteronuclear spins. These methods were applied to investigate the heteronuclear relaxation properties of 113Cd in 113Cd2LAC9(61), a Cys6Zn2 DNA-binding domain. Data from the experiments confirm biochemical evidence that more significant differences occur in the metal-protein interactions between the two metal-binding sites than has been previously identified for proteins containing this motif.     

Glomerular permeability: in vivo tracer studies with polyanionic and polycationic ferritins

The influence of molecular charge on glomerular permeability to the globular ferritin molecule in vivo was investigated. Mice (Charles River CD strain) and rats (Munich-Wistar strain) were injected intravenously either with native anionic ferritin or various cationized derivatives with different isoelectric points (pI) and the kidneys were examined by electron microscopy. Native anionic ferritin was almost completely restricted from entry into the glomerular filter at the level of the endothelium and subendothelial layer of the glomerular basement membrane (GBM). Cationized derivatives penetrated the filter in increasing amounts depending on the pI of the tracer. Regardless of charge, all molecules that filtered through the lamina densa of the GBM and reached the subepithelial layer were completely restricted from entry into the urinary space at the level of filtration slits and appeared in phagosomes present in podocytes. Reduction of arterial pressure of cessation of renal blood flow did not influence the movement of ferritin molecules into the GBM. The results are consonant with physiological studies indicating charge dependent restriction of polyanion transport by the mammalian glomerulus. These tracer studies, in conjunction with cytochemical and biochemical evidence for the presence of polyanionic glycoproteins in the glomerular filter, suggest that glomerular restriction of plasma proteins occurs in part by a process similar to that which exludes negatively charged macromolecules in polyanionic gel systems.     

Increased prolactin and thyrotrophin secretion following oral metoclopramide: dose-response relationships

In pregnant female rat, oxygen tension was measured in vivo with an oxygen microelectrode and the following statistically significant data (Student's test) were obtained: -- not significant variability in four groups of six control rats; -- highly significant decrease of oxygen tension twenty-four hours after biovariectomy in four groups of six operated rats; -- in twelve operated and treated by substitutive hormonotherapy rats, pO2 was at the same level than in control rats; -- in eighteen operated rats, the oxygen tension measured after embryonic death was identical to control rats. These experiments clearly demonstrate twenty four hours after biovariectomy a decreased oxygen tension. Simultaneously to this decrease, a diminution of uterine blood flow takes place. This pO2 diminution should be dependent on decrease of ovarian hormones since the substitutive hormonotherapy prevents its appearance. A good explanation of this phenomenon is the high requirement of hypoxic embryo for oxygen; moreover after the embryonic death, the intra-uterine pO2 increases.     

Resolution of focal fatty infiltration of the liver

Focal fatty infiltration of the liver has been recognized as a distinct entity only since 1980. It may simulate neoplastic or other low-density parenchymal lesions, including abscesses and hemangiomas. I report a case of histologically confirmed focal fatty infiltration of the liver in a man with obesity and alcoholism; the lesion disappeared after the patient began a program of exercise, weight loss, and abstinence from alcohol. The possible pathogenesis of focal fatty infiltration of the liver is reviewed, diagnostic techniques are described, and reversibility of the lesion is discussed.     

Molecular typing shows a high level of HLA class I incompatibility in serologically well matched donor/patient pairs: implications for unrelated bone marrow donor selection

In comparison with HLA-matched sibling bone marrow transplants, unrelated donor transplants are associated with increased graft-versus-host disease and graft failure. This is likely in part due to HLA incompatibilities not identified by current matching strategies. High resolution DNA-based typing methods for HLA class II loci have improved donor selection and treatment outcome in unrelated donor bone marrow transplantation. By using DNA-based typing methods for HLA-A and -B on a cohort of 100 potential bone marrow donor/patient pairs, we find that serological typing for HLA class I is limited in its ability to identify incompatibilities in unrelated pairs. Furthermore, the incompatibilities identified are associated with the presence at high frequency of alloreactive cytotoxic T-lymphocyte precursors. DNA typing also indicates that HLA-C mismatches are common in HLA-A and -B serologically matched pairs. Such mismatches appear to be significantly less immunogenic with respect to cytotoxic T-lymphocyte recognition, but are expected to influence natural killer cell activity. Thus, improved resolution of HLA class I shows many previously undisclosed mismatches that appear to be immunologically functional. Use of high resolution typing methods in routine matching is expected to improve unrelated donor selection and transplant outcome.