T cells specific for alpha-beta interface regions of hemoglobin recognize the isolated subunit but not the tetramer and indicate presentation without processing.

Processing of a protein antigen into fragments is believed to be a prerequisite for its presentation by the antigen-presenting cell to the T cell. This model would predict that, in oligomeric proteins, T cells prepared with specificity for regions that are buried within subunit association surfaces should recognize the respective regions in vitro equally well on the isolated subunit or on the oligomer. Three hemoglobin (Hb) alpha-chain synthetic peptides, corresponding to areas that are situated either completely [alpha-(31-45)] or partially [alpha-(41-45) and alpha-(81-95)] within the interface between the alpha and beta subunits of Hb, and a fourth peptide representing a completely exposed area in tetrameric Hb were used as immunogens in SJL/J (H-2s) mice. Peptide-primed T cells were passaged in vitro with the respective peptide to obtain peptide-specific T-lymphocyte lines. T-cell clones were isolated from these lines by limiting dilution. T-cell lines and clones that were specific for buried regions in the subunit association surfaces recognized the free peptide and the isolated subunit but not the Hb tetramer. On the other hand, T cells with specificity against regions that are not involved in subunit interaction and are completely exposed in the tetramer recognized the peptide, the isolated subunit, and the oligomeric protein equally well. The responses of the T-cell lines and clones were major histocompatibility complex-restricted. Since the same x-irradiated antigen-presenting cells were employed, the results could not be attributed to differences or defects in Hb processing. The findings indicate that in vitro the native (unprocessed and undissociated) oligomeric protein was the trigger of major histocompatibility complex-restricted T-cell responses.     

Tumor inhibiting properties of stereoisomeric [1,2-bis(3-hydroxyphenyl) ethylenediamine]dichloroplatinum(II)-complexes, Part II: Biological properties

The activity of stereoisomeric [1,2-bis(3-hydroxyphenyl)ethylenediamine] dichloroplatinum(II)-complexes (1-PtCl2,R,S; 2-PtCl2, R,R/S,S; 3-PtCl2, R,R; 4-PtCl2, S,S) on several tumor models (MDA-MB 231 breast cancer cell line; P 388 leukemia, mouse; L 1210 leukemia, mouse; L 5222 leukemia, rat; Ehrlich ascites tumor, mouse--wildtype; cisplatin-, etoposide-, cyclophosphamide-, and daunomycin-resistant, resp.) is described. For comparison the analogous [1,2-bis(4-hydroxyphenyl)ethylendiamine]dichloroplatinum (II)-complexes (5-PtCl2, R, S; 6-PtCl2, R,R/S,S; 7-PtCl2, R,R; 8-PtCl2, S,S) and cisplatin are used. 1-PtCl2 to 4-PtCl2 (OH in 3,3'-positions) show their maximum antitumor effect at lower doses than 5-PtCl2 to 8-PtCl2 (OH in 4,4'-positions). 2-PtCl2 and 6-PtCl2 (R,R/S,S) are more active than 1-PtCl2 and 5-PtCl2 (R,S). 4-PtCl2 and 8-PtCl2 (S,S) are superior to 3-PtCl2 and 7-PtCl2 (R,R). On the L 5222 leukemia 2-PtCl2 (R,R/S,S), 4-PtCl2 (S,S) and 8-PtCl2 (S,S) markedly surpass cisplatin. Strong effects are produced by 2-PtCl2 to 4-PtCl2 on the Ehrlich ascites tumor (wildtype, cisplatin-, etoposide-, cyclophosphamide-, and daunomycin-resistant, resp.). The combination of 4-PtCl2 with cisplatin results in a weakly synergistic effect.     

Nucleoside analogues: 8. Some isomers of B.3839, the original 5-fluorouracil/nitrosourea molecular combination, and their effect on colon, breast and lung tumours in mice

This study describes the manipulation of secondary products arising from the synthesis of the prototypical molecular combination of 5-fluorouracil (5-FU) and chloroethylnitrosourea (CNU), B.3839, in order to investigate the effects produced by connecting the C-S-C-C-CNU chain to the 5-FU ring in different ways. The isolation of phthalimide precursors of these compounds and the transformation into CNUs is described. Anti-tumour activity of these molecular combinations against a series of experimental murine colon, lung and mammary tumours is presented. The spectrum of anti-tumour activity displayed is interesting but defies simple explanation without further detailed in vivo pharmacokinetic and metabolism studies in order to define optimal profiles for activity.     

Probe tine diameter and probing depth

The aim of this study was to assess the effect of difference in tine diameter on probing pocket depth measurement. 2 sets of tines with Williams markings at 1, 2, 3, 5, 7, 8, 9 and 10 mm, and with a "round" tip, diameter 0.5 mm, were compared. One set was described as parallel-sided, the other as tapered. The parallel-sided tine was almost parallel from the 10 mm marking to the tip (tip diameter mean = 0.46 mm, 95% C.I. 0.456-0.464), while the corresponding diameter for the tapered tine varied (tip diameter mean = 0.48 mm, 95% C.I. 0.473-0.489). Calibration markings appeared highly consistent with the expected value to within 0.01 mm. The tines were mounted in Brodontic handles at 0.25 N. Examiner probing repeatability yielded kappa 0.86 for "parallel-sided" and 0.81 for "tapered" tines in vivo. 412 approximal pockets were assessed in 53 patients with routine chronic adult periodontitis, mean age 42.1 years. Each site had a probing depth of greater than or equal to 5 mm, PlI less than or equal to 1, GI less than or equal to 1, PBI less than or equal to 1. Each site was probed 2x with a 15-min interval. At the first 251 sites, the parallel-sided tine was used initially, and the tapered at the remaining 161 sites. Results indicated a highly significant tendency for the parallel-sided tine to yield a deeper reading when a difference occurred. These findings indicate that with adequate training providing high examiner repeatability, one source of error in probing data can be minimised.     

Discovery and implications of the immunogenicity of free small synthetic peptides: powerful tools for manipulating the immune system and for production of antibodies and T cells of preselected submolecular specificities

Recent studies from this laboratory have found, contrary to a long-held belief, that synthetic peptides of a protein, as small as six residues, when immunized in their free form (i.e., without coupling to any carrier), elicit the formation of antibodies with submolecular binding specificities to preselected protein regions. These peptides could represent either the antigenic sites of the protein or surface regions that are not immunogenic when the intact protein is the antigen. In either case, the antibodies bind specifically to the intact protein, exclusively at the region used in immunization. Monoclonal antibodies with preselected specificities can also be produced by hybridoma technology from the spleens of the immunized animals. Furthermore, free synthetic peptides have been employed to generate T-cell lines and T-cell clones with specificities to preselected immunogenic locations and also to induce tolerance to such preselected locations. These fascinating breakthroughs indicate that free synthetic peptides may be used in the future as powerful tools in basic investigations and in therapeutic and diagnostic applications.     

Time-dependent study of the antibody response to sperm-whale myoglobin: recognition of the antigenic sites is unaltered over an extended period of immunization

We have recently shown that the antigenic structure of sperm-whale myoglobin is not dependent on the host species in which the antisera are raised. The purpose of the present work was to determine whether or not the molecular immune recognition of a protein by antibody is subject to a time-dependency. We have investigated the recognition of the antigenic sites by serial antisera obtained in two rabbits at different times after the initial immunization, from the earliest bleeding with detectable antibodies (9 days) up to a year. The specificity of these antisera was determined by their cross-reactions with 13 myoglobins from different species. The reactivity was measured by quantitative immunoadsorbent titration studies from the amount of radioiodinated antibodies that could be bound maximally (i.e. plateau binding values) by immunoadsorbents of each myoglobin. From these studies and our recent structural analysis, which enabled us to identify for each of these myoglobins the regions retaining reactivity with antibodies to sperm-whale myoglobin, we have concluded that following maturation of the immune response the antigenic recognition of a protein (i.e. the regions that are recognized as antigenic) is not dependent upon the time antisera are obtained after the first immunization. In the early periods of the response, fluctuations are observed in the relative immunodominance of the sites. This further confirms our earlier conclusions that the antigenic sites on a protein are structurally inherent in the protein.     

Vaccination with a MHC class II peptide in Alum and inactive pertussis strongly ameliorates clinical MG in C57BL/6 mice

We have investigated the efficacy of immunization against peptides from predisposing MHC class II molecules in human-compatible adjuvants for ameliorating experimental autoimmune myasthenia gravis (EAMG). C57BL/6 mice were immunized three times with the peptide I-Abetab62-76 in Alum+killed pertussis organisms (PT) prior to two injections with tAChR. The treatment greatly reduced the occurrence and severity of clinical MG relative to controls that received saline/Alum+PT or none. It also reduced antibody and T-cell responses against tAChR. The results have important implications for the possible immunotherapy of MG by targeting disease-associated MHC.     

Value of power doppler and 3D vascular sonography as a method for diagnosis and staging of prostate cancer

OBJECTIVES: To compare the value of Power Doppler Sonography (PDS) and B mode sonography in the diagnosis of prostate cancer and to assess the value of PDS to specify capsular effraction of the cancer. PATIENTS AND METHODS: 323 patients were investigated: 41 control subjects allowed the establishment of normal vascular semiology and 282 patients with suspected cancer (PSA >4ng/ml). Power Doppler Sonography with 3D reconstruction was used to describe Power Doppler Sonography features of normal or abnormal vessels. Three types of blood supply(a: regular avascular posterior peripheral margin, b: irregular avascular posterior peripheral margin, c: vessels crossing the posterior peripheral margin) were described as a function of the presumed stage of cancer (a: intraprostatic, b: undetermined, c: extraprostatic). Comparison with histology was performed on random biopsies without Doppler (282 cases) (median PSA level = 15.8ng/ml), on second biopsies indicated with PDS (72 cases), and radical prostatectomy specimens (63 cases). RESULTS: A cancer was diagnosed in 157 of the 282 patients (55.7%) with suspected cancer. The overall sensitivity of PDS in the initial diagnosis of prostatic cancer was 92.4% and its specificity was 72% (versus 87.9% and 57.6% for sonography alone respectively). The negative predictive value of PDS was elevated to 80.6% (p<0.0001). Targeting area presenting abnormal blood flow in any part of the prostate was useful to detect isoechoic or lesions in patients with first negative biopsy results (in 41 of 72 targeted patients with first negative biopsies with PDS a cancer was diagnosed: 58% of these cancers had less than 3 positive biopsies and 34% only one positive biopsy). The 3 vascular types a, b, c were evaluated prospectively in the detection of capsular effraction. The presence or absence of vessels crossing the capsule to determine an extracapsular extension was a significant sign (p<0.0001). Capsular effraction was detected in 3 of the 27 cases (11%) of type a cancer and in 16 of the 18 cases (87%) of type c cancer. CONCLUSION: PDS improves the accuracy of echographic imaging in the diagnosis of cancer. Combining first sextant biopsies and targeted areas presenting abnormal blood flow using PDS can increase cancer detection with an optimized number of biopsy cores. The risk of extracapsular involvement can be evaluated by the presence of vessels perforating the capsule.     

A pilot trial of RNS60 in amyotrophic lateral sclerosis

Introduction: RNS60 is a novel immune-modulatory agent that has shown neuroprotective effects in amytrophic lateral sclerosis (ALS) preclinical models. RNS60 is administered by weekly intravenous infusion and daily nebulization. The objective of this pilot open-label trial was to test the feasibility, safety, and tolerability of long-term RNS60 administration in ALS patients. Methods: The planned treatment duration was 23 weeks and the primary outcomes were safety and tolerability. Secondary outcomes included PBR28 positron emission tomography (PET) imaging and plasma biomarkers of inflammation. Results: Sixteen participants with ALS received RNS60 and 13 (81%) completed 23 weeks of RNS60 treatment. There were no serious adverse events and no participants withdrew from the trial due to drug-related adverse events. There were no significant changes in the biomarkers. Discussion: Long-term RNS60 administration was safe and well-tolerated. A large, multicenter, phase II trial of RNS60 is currently enrolling participants to test the effects of RNS60 on ALS biomarkers and disease progression. Muscle Nerve 59 :303–308, 2019     

Prediction of Caesarean Section from Ultrasound and Clinical Assessment of Fetal Size

EDITORIAL COMMENT: We accepted this paper for publication because the authors have explored the possible value of ultrasound versus clinical assessment of fetal size to see whether they can predict the need for Caesarean section when the baby is large. In this study both the ultrasound and the clinical assessment were impressively accurate and fetal femur length was the most accurate of the ultrasound parameters in assessing fetal weight in predicting the need for Caesarean section. However, as the authors indicate, none of these methods of assessment of fetal size are recommended as an absolute indication for Caesarean section. Every obstetrician uses clinical assessment of the fetus when evaluating management of the patient but the need for clinical judgment remains.
Summary: One hundred and five women with singleton pregnancies and cephalic presentation were assessed. Fundal height and a clinical estimate of fetal weight were recorded. The fetal biparietal diameter, abdominal circumference and femur length were measured with ultrasound. Ultrasound estimated fetal weight was calculated using 3 different formulas (Shepard, Campbell and CUHK). The liquor volume was assessed using the amniotic fluid index.
Ultrasound was able to predict Caesarean section with more reliability than clinical assessment of fetal size or weight. The biparietal diameter, fundal height and amniotic fluid index were poor predictors of mode of delivery. The measurements which best predicted the mode of delivery were the fetal femur length and abdominal circumference. Femur length, but not abdominal circumference, was a statistically better predictor of Caesarean section than clinical estimation of fetal weight. There was no improvement in prediction using ultrasound estimated fetal weight.