Detection of radiolabelled bone marrow cells bearing surface ace immunoglobulins by combined autoradiography and immunoperoxidase

A method is described for the simultaneous detection of radiolabelled bone marrow cells bearing surface immunoglobulins by combined autoradiography and immunoperoxidase. Bone marrow cells from normal CBA mice prelabelled in vivo with ¹²⁵IUDR or exposed in vitro to [³H]thymidine were incubated with rabbit anti-mouse immunoglobulins under capping conditions, washed, cytocentrifuged and treated with methanol and hydrogen peroxide to destroy endogenous peroxidase. Cells were then covered with peroxidase-conjugated goat anti-rabbit immunoglobulins, washed, treated with diaminobenzidine a and hydrogen peroxide and finally covered with autoradiographic stripping film and exposed for different times. Peroxidase-positive cells were typically capped and those radiolabelled had autoradiographic silver grains overlying the nucleus.     

Retinal ganglion cells that project to the superior colliculus and pretectum in the macaque monkey

Horseradish peroxidase was injected into the superior colliculus or pretectum or both in order to label, by retrograde axoplasmic transport, the retinal ganglion whose cells axons innervate the dorsal midbrain. The dendrites of ganglion cells were sufficiently well-labelled to reveal their overall morphological characteristics. It was therefore possible to compare the number and form of ganglion cells projecting to the midbrain with the total population of ganglion cells as revealed by optic nerve injections, and with ganglion cells labelled by injections in the lateral geniculate nucleus. We found that not more than 10% of all retinal ganglion cells project to the superior colliculus in the macaque monkey. This percentage varies little over the retina, being about 6% of all ganglion cells near the fovea and increasing slightly with eccentricity. The superior colliculus does not receive a projection from P beta cells and only a few P alpha cells terminate there. The majority of cells which project to the superior colliculus have a small- to medium-sized cell body and sparsely branched dendritic tree. We have called them P gamma and P epsilon cells by analogy with the gamma cells and epsilon cells in the cat's retina. Anatomically the P gamma and P epsilon cells are heterogeneous, which would be consistent with the physiological heterogeneity found for ganglion cells which project to the midbrain in monkeys.     

Compositional and metabolic heterogeneity of alpha 2- and beta-very-low-density lipoproteins in subjects with broad beta disease and endogenous hypertriglyceridemia

The catabolism of alpha 2- and beta-very-low-density lipoproteins (VLDL) was studied in normolipidemic and hyperlipidemic subjects to determine whether differences in the catabolism of these subfractions are due to their composition. alpha 2-VLDL (cholesterol/triglyceride ratio, 00.18 +/- 0.06; and apoprotein E/C ratio, 0.27 +/- 0.22, n = 4) and beta-VLDL (cholesterol/triglyceride ratio, 0.67 +/- 0.13; and apoprotein E/C ratio, 1.05 +/- 0.52, n = 4) were isolated from subjects with broad beta disease, iodinated, and injected in five normolipidemic subjects, six with broad beta disease, and five with endogenous hypertriglyceridemia. VLDL, intermediate (IDL) and low-density lipoprotein (LDL) apoprotein (apo)-B radioactivity (tetramethylurea insoluble) following injection of 125I-labeled alpha 2- and beta-VLDL decayed biphasically in all subjects, and this decay in normolipidemic subjects was more rapid than in subjects with broad beta disease (P = 0.004) or endogenous hypertriglyceridemia (P = 0.004 for alpha 2- and P = 0.010 for beta-VLDL). The residence times, however, for the delipidation chain in alpha 2-VLDL were similar in all the subjects and varied from three to six hours. The decay of radioactivity in beta-VLDL in subjects with broad beta disease was much slower (residence time, 36.9 +/- 24.4 hr, n = 7) than in normolipidemic subjects (residence time, 7.56 +/- 4.6 hr, n = 5) or in subjects with endogenous hypertriglyceridemia (residence time, 10.6 +/- 4.65, n = 4). The residence time for alpha 2-VLDL was longer than for beta-VLDL in all subjects, suggesting that alpha 2-VLDL is a precursor to beta-VLDL. To test this directly, iodinated alpha 2-VLDL was injected into a subject with broad beta disease and the radioactivity in the subfractions was followed. The radioactivity from alpha 2-VLDL was transferred into beta-VLDL supporting, the notion that alpha 2-VLDL generated some beta-VLDL. Nicotinic acid treatment of a subject with broad beta disease accelerated the catabolism of alpha 2- and beta-VLDL without changing the VLDL composition.     

T cell subsets and cellular immunity in end-stage renal disease

The T lymphocyte population was studied by immunofluorescent staining with monoclonal antibodies and laser flow cytometry in the blood of 50 patients with end-stage renal disease undergoing long-term maintenance intermittent hemodialysis. The absolute number of T cells was lower in patients receiving dialysis for more than one year (p less than 0.001), as was the absolute count of helper T cells (p less than 0.005). In patients under 30 years of age, the absolute number of helper T cells was markedly reduced, whereas the number of suppressor/cytotoxic T lymphocytes was not changed. In patients between the ages of 30 and 60 years, both helper and suppressor cells were significantly reduced. In patients over 60 years of age, only the number of helper T cells was reduced. The in vitro response of patients' lymphocytes was reduced both in the mixed lymphocyte reaction (p less than 0.01) and after phytohemagglutinin stimulation (p less than 0.001). Natural killer cytotoxicity of patients' peripheral blood mononuclear cells, however, was unaffected.     

Initiation of two distinct forms of atrioventricular nodal reentrant tachycardia during programmed ventricular stimulation in man

Of 42 patients with supraventricular tachycardia related to dual atrioventricular (A-V) nodal pathway conduction, 8 had sustained tachycardia induced during programmed ventricular stimulation. The characteristics of the tachycardia in three patients suggested that the A-V nodal reentrant tachycardia used a slow pathway for anterograde conduction and a fast pathway for retrograde conduction (slow-fast form). In these patients, the retrograde effective refractory period was longer in the slow than in the fast pathway. Ventriculoatrial (V-A) conduction curves (V₁-V₂, A₁-A₂) were smooth. Ventricular premature beats, being conducted retrograde over the fast pathway, could activate the slow pathway in an anterograde direction, initiating the slow-fast form of A-V nodal reentrant tachycardia. In the remaining five patients, the tachycardia used a fast pathway for anterograde conduction and a slow pathway for retrograde conduction (fast-slow form). In these patients, the retrograde effective refractory period was longer in the fast than in the slow pathway. V-A conduction curves (V₁-V₂, A₁-A₂) could be either smooth or discontinuous if there was a sudden increase in V-A conduction time. Ventricular premature beats, conducted retrograde over the slow pathway, could activate the fast pathway in an anterograde direction, establishing a tachycardia circuit in reverse of the slow-fast form. In both groups of patients, the ventricular pacing cycle length appeared to be a crucial factor in the ability to expose functional discordance between the two A-V nodal pathways during retrograde conduction.The fast-slow form of A-V nodal reentrant tachycardia, similar to the slow-fast form, could also be induced during atrial premature stimulation in two patients. In this situation, the slow pathway having an anterograde effective refractory period longer, than that of the fast pathway was a requisite condition; anterograde A-V nodal conduction curves (A₁-A₂, H₁-H₂) were smooth. Atrial premature beats, conducted anterograde over the fast pathway, could activate the slow pathway in a retrograde direction resulting In an atrial echo or sustained fast-slow form of A-V nodal reentrant tachycardia.     

Long-term efficacy and urological toxicity of low-dose-rate brachytherapy (LDR-BT) as monotherapy in localized prostate cancer

PurposeThe purpose of this study was to evaluate the incidence of late severe (≥Grade 3) urinary toxicity and the long-term efficacy after low-dose-rate brachytherapy (LDR-BT) in patients with localized prostate cancer (PCa).Methods and MaterialsDuring the years 1999–2008, 241 patients with PCa who underwent LDR-BT with I¹²⁵ and were followed up in Kuopio University Hospital were included to this analysis. The incidence of late severe (Grade 3) urinary toxicity and the long-term efficacy results were analyzed.ResultsAll D'Amico risk groups were represented, as 58.9%, 35.3%, and 5.8% of the patients were classified as low-, intermediate-, and high-risk patients, respectively. With a median followup of 11.4 years after implantation, the incidence of severe urinary toxicity increased throughout the followup period. The risk of Grade 3 urinary toxicity was highest among patients with higher Gleason scores (p = 0.016) and higher initial urine residual volumes (p = 0.017) and the cumulative incidence of severe urinary toxicity was 10.0%. The crude rate for transurethral prostatic resection was 5.8%. The relapse-free survival, the cause-specific survival, and the overall survival were 79.3%, 95.0%, and 66.4%, respectively.ConclusionsThe treatment was well tolerated as 90% of patients avoided any Grade 3 urinary toxicity. LDR-BT for localized PCa achieved high and durable efficacy. These results support the role of LDR-BT monotherapy as one of the valid primary treatment options for low-risk and favorable intermediate-risk patients.     

Implementation of an HDR brachytherapy–based breast IORT program: Initial experiences

PurposeA multidisciplinary team at our institution developed a novel method of intraoperative breast radiation therapy (precision breast intraoperative radiation therapy [PB-IORT]) that uses high-dose-rate brachytherapy with CT on-rails imaging to deliver high-dose, customized radiotherapy to patients with early-stage breast cancer. This report summarizes our program's experience developing and implementing PB-IORT.Methods and MaterialsLiterature on PB-IORT was reviewed including published articles and abstracts. To evaluate case volume, all patients with a breast cancer diagnosis who underwent breast surgery or breast radiation (2010–2017) at our academic institution were identified. Patients were stratified into pre-IORT and post-IORT eras with initiation of our PB-IORT program in October 2013. Overall trends in surgical and radiation therapy volume in each era were analyzed by linear regression. Travel distance for all surgical patients was calculated using Google Maps (Alphabet Inc.) and then compared between IORT and non-IORT patients.ResultsData from a PB-IORT Phase 1 trial found that the primary endpoints were met and that PB-IORT is feasible and safe. The direct health system's delivery costs for PB-IORT exceed those of 16-fraction whole-breast irradiation when accounting for consumable supplies (multilumen balloon applicator = $2,750 per patient). There was a significant increase in yearly growth of breast cancer surgical volume with PB-IORT.ConclusionsAccrual rates for the ongoing Phase II trial have been quicker than expected in an area where more research is needed. The rapid accrual indicates patient interest and demand for this treatment and that it is very feasible to get more data from randomized trials.