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Background and Objectives Pathogen reduction technology using amustaline (S-303) was developed to reduce the risk of transfusion-transmitted infection and adverse effects of residual leucocytes. In this study, the viability of red blood cells (RBCs) prepared with a second-generation process and stored for 35 days was evaluated in two different blood centres. Materials and Methods In a single-blind, randomized, controlled, two-period crossover study (n = 42 healthy subjects), amustaline-treated (Test) or Control RBCs were prepared in random sequence and stored for 35 days. On day 35, an aliquot of 51Cr/99mTc radiolabeled RBCs was transfused. In a subgroup of 26 evaluable subjects, 24-h RBC post-transfusion recovery, mean life span, median life span (T50) and life span area under the curve (AUC) were analysed. Results The mean 24-h post-transfusion recovery of Test and Control RBCs was comparable (83·2 ± 5·2 and 84·9 ± 5·9%, respectively; P = 0·06) and consistent with the US Food and Drug Administration (FDA) criteria for acceptable RBC viability. There were differences in the T50 between Test and Control RBCs (33·5 and 39·7 days, respectively; P 0·05). Following infusion of Test RBCs, there were no clinically relevant abnormal laboratory values or adverse events. Conclusion RBCs prepared using amustaline pathogen reduction meet the FDA criteria for post-transfusion recovery and are metabolically and physiologically appropriate for transfusion following 35 days of storage.

Understanding blood volume changes in children with malaria is important for managing fluid status. Traditionally, blood/red cell volume measurements have used radioactive chromium isotopes. We applied an alternative approach, using non-radioactive chromium-53 labelling and mass spectrometry to investigate red cell volume (RCV) in Gabonese children with malaria. Nineteen children with malaria participated (10 severe, 9 moderately severe; ages 15 months to 7 years). Blood labelled with 53Cr-chromate exvivo was re-injected, then sampled 30 min later. Pre- and post-injection 53Cr content were measured by gas chromatography/electron ionisation mass spectrometry of the chromium-trifluoroacetylacetone (TFA) chelate, calibrated against 50Cr standards. Blood and red cell volumes were calculated from isotopic dilution in 15 of 19 children (in four, insufficient signal mitigated analysis). In this small pilot study, there were no significant differences between moderate and severe cases. Including all subjects, the mean RCV was reduced compared with predicted values (184 vs. 269 mL; p = 0.016) but blood volume, 71 ± 33 mL/kg (normalised for weight), was close to predicted, ∼77 mL/kg, commensurate with reduced haematocrit. Blood lactate concentration correlated negatively with RCV/weight (r = −0.56, p = 0.028), consistent with anaemia. In one case, sequential samples over 42 days gave an estimated rate of 53Cr disappearance of 1.4%/day (equivalent half-life: 70 days). 53Cr-labelling of red cells may be used to estimate blood and red cell volumes and can be used as an investigative tool in situations such as childhood diseases and resource-constrained settings. Although the red cell mass is depleted in malaria, the blood volume appears relatively well preserved. Copyright © 2009 John Wiley & Sons, Ltd.

The intravascular clearance of intravenously administered endotoxin prior to and after administration of immune serum has been compared in untreated rabbits and in rabbits pretreated with warfarin sodium for 4 and 66 days, respectively. Firstly, the present observations show that the phagocytic rate of endotoxin without immune serum is significantly higher in rabbits exposed to “short-term” warfarin pretreatment than in those exposed to “long-term” pretreatment and in untreated rabbits. The “long-term” warfarin treatment is supposed to depress the function of the reticuloendothelial system, particularly in the liver. Secondly, the phagocytic rate of endotoxin increases significantly following the administration of immune serum, which almost completely abolishes the difference between the phagocytic rates of the groups prior to administration. Administration of immune serum seems to decrease the uptake of endotoxin in some organs or provoke an immediate redistribution of the endotoxin between the organs. The warfarin pretreatment influences the relative weight of the liver, and the duration of the warfarin pretreatment seems to influence the organ localization of endotoxin.

The sites of destruction of 51Cr tagged G-6-PD deficient red cells in normal and primaquine sensitive Sardinians after drug administration have been studied by means of surface monitoring and, in one case, by direct quantitation after surgery. In practically all cases a liver to spleen ratio of one could be recorded, with a constant splenic participation in the uptake of radioactivity. It was observed that, beside the major role played by the liver in the sequestration and disposal of severely damaged G-6-PD deficient red cells after administration of primaquine, also the spleen has a significant share in the disposal of damaged cells.

To 11 patients with sickle cell anaemia pharmacological doses of androgens were given for periods varying between 3 and 16 weeks. To 3 of the patients two courses of treatment were given. During the treatment 10 of the patients showed an increase of the total red cell mass varying between 25 and 125% of the control value. One patient did not respond. The treatment had no discernible effect on the sickling tendency of the erythrocytes at different levels of oxygen tension and there was no consistent alteration of the red cell survival measured with radioactive chromium. Weight gain was seen in all patients and hirsutism was seen in most of the females. In spite of the marked increase in red cell mass there was no certain increase in the frequency of crisis among the female patients. Priapism occurred in all of the adult males responding to the therapy. The number of patients studied is, however, small and the observation time too short to allow any firm conclusion on the effect of androgen treatment on the frequency of vaso-occlusive crisis.

3 patients with myelosclerosis have subsequently developed paroxysmal nocturnal haemoglobinuria, with positive in-vitro lysis tests (acidified-serum lysis, cold-antibody lysis and sucrose lysis), characteristic of PNH. These cases provide further evidence of a relationship between PNH and myeloproliferative disorders; they support the proposition that PNH is an acquired defect of erythropoiesis due to an abnormal clone. In two of the patients PNH has been severe with marked intravascular haemolysis, so that in these cases the clinical course and management of the original disease have been influenced by the PNH. In the third patient the PNH defect has, so far, been essentially only a phenomenon of in-vitro tests and apparently without clinical effect.

Two sisters had a congenital hypochromic anaemia in spite of hyperferraemia with a fully saturated serum transferrin, and in spite of heavy iron deposits in the liver. Bone marrow iron was reduced. The anaemia was markedly increased during pregnancy, when each of the sisters needed 8 blood transfusions. Erythrokinetic studies were compatible with hypoplastic anaemia. The bone marrow, however, was hyper-cellular with an increased erythropoiesis. Our patients resembled the two patients described by Shahid & Diamond (1964). It is postulated that the cause of the disease is a congenital defect in the one-way transport mechanism for iron from transferrin and into the erythroid cell.

The erythrocyte sequestration in the spleen and liver was investigated in 25 cases of immunohaemolysis by surface counting after infusion of 51Cr-labelled autologous erythrocytes. The erythrocyte sequestration pattern was found to depend upon the treatment with prednisone given at the time of the investigation and furthermore upon the total haemolytic intensity. Increased sequestration of erythrocytes in the liver is always present in cases of severe hyperhaemolysis. After prednisone treatment, hypersequestration of erythrocytes in the spleen predominates. In the present study increased sequestration in the liver was even demonstrated in about 50 % of the cases in which incomplete non-complement-fixing antibodies were found. In the majority of cases prednisone therapy gave only short-lasting remission. Frequently, it is difficult to reduce the dose of prednisone because of increasing haemolytic intensity. Complete, long-lasting remission seems to follow splenectomy in cases characterized by a splenic sequestration pattern during periods without prednisone therapy, but if hepatic hypersequestration has been demonstrated the remissions are not complete. The need for steroids decreased after splenectomy in practically all cases. A combination of prednisone therapy and early splenectomy is advisable in most cases of immunohaemolysis.

A case of infectious mononucleosis with longstanding thrombocytopenia, not responsive to corticosteroid therapy, is presented. Rapid normalization of the platelet level was revealed by splenectomy. The patient is the fourth known case in which it has been necessary to perform splenectomy. A study made with 51Cr-labelled homologous platelets showed a very short half-life, along with a high spleen/liver ratio of the activity. These examinations have not previously been performed in this condition.

Platelet kinetics, volume of megakaryocytes per μl sternal bone marrow, and the volume of the spleen have been quantitated in 20 cases of untreated polycythaemia vera (PV). There was a very good relationship between platelet production rate and megakaryocyte volume/μl bone marrow. The results suggest that megakaryopoiesis in PV represents mainly an effective platelet production. The megakaryocyte volume/μl bone marrow was significantly related to the body platelet mass (p < 0.001), but its relationship to the peripheral platelet count was significant only at the 10 % level. This was due to altered platelet distribution caused by differences in spleen and blood volumes. There was a negative relationship between platelet half-life time (T 1/2) and spleen volume (0.10 > p > 0.05) suggesting that enlargement of the spleen as such influences platelet survival. A highly significant correlation between platelet production rate and spleen volume, as well as between body platelet mass and spleen volume was found. The influence of the spleen size on platelet kinetics in PV has been discussed.

45 patients with chronic non-specific obstructive lung disease had a whole body/venous haematocrit ratio of 0.86 ± 0.008 SEM. This is lower than the ratio found in normal adults. For greatest accuracy independent measurement of red cell and plasma volumes should be undertaken in this condition.

The relationship between the exchangeable splenic platelet pool, determined by recovery of infused 51Cr-labelled platelets, and the spleen size, determined by 99mTc scintigraphy, has been investigated in 22 cases of polycythaemia vera and 6 control subjects. The results have shown a highly significant negative correlation, indicating that with increasing splenomegaly a larger proportion of the body platelet mass is pooled in the spleen. A scintigraphic determination of the spleen size gives a rough estimation of the splenic platelet pool.

A simple analytical model has been worked out for quantitative assessment of temporary shifts in erythrocyte survival curves after labelling with radioactive tracers. This model was applied to 51Cr and DF32P curves from rabbits exposed to experimental fever induced by either bacterial pyrogen, heated milk or external heating in a climatic chamber. Thereby a quantitative expression of the fever-induced erythrocyte destruction was obtained. In all three methods for elevating the body temperature experimentally the studies showed an increased loss of activity during the fever, either in terms of the slope of the curve segment concerned with fever compared to the pre-or post-febrile periods or in terms of the deviation of the observed curves from the expected, spontaneous course in the rabbit. The maximum loss occurred soon after the initiation of fever, and prolongation of the fever period did not cause a continued increase in the destruction of the labelled erythrocytes. A certain correlation was found between the initial action of heat and the magnitude of the erythrocyte destruction. After the cessation of a short-lasting period of fever the tracer curve again approached the expected spontaneous course, explained by an age-dependent variation in the heat susceptibility of the erythrocytes. By using the age-distribution function for rabbit erythrocytes, an explanation of the demonstrated differences in the simultaneous changes in radioactivity and haemoglobin or PCV is attempted. It is concluded that elevated body temperature shortens the mean survival time of the red cells. The average reduction of the red cell mass in the present study was about 15% and the maximum loss about 35%.

Eleven patients with essential hypertension were treated with hydrochlorothiazide/KCl for three months. A significant fall in total blood volume, exchangeable sodium and blood pressure could be demonstrated both after one to two weeks and after three months of treatment, while a fall in serum sodium and body weight was only significant after one to two weeks of treatment. Thus, hypovolemia may, perhaps, be the explanation for the antipressor effect of thiazide even after three months of treatment.

Erythrocyte pooling and sequestration in the spleen were studied in 73 patients with splenomegaly due to various haematological disorders using 51Cr-labelled autologous erythrocytes and surface counting. Arbitrary values for the accumulation of activity in spleens of different sizes were obtained by correcting measured splenic surface activities (correction factors have been experimentally estimated). Absolute values for the splenic content of 51Cr and 125I were obtained by comparative surface activity measurements over the enlarged spleen and a model of the spleen charged with a known amount of activity. In this way the splenic erythrocyte and plasma volume may be estimated. In cases of pronounced splenomegaly without complicating immunohaemolysis the splenic erythrocyte pool (splenic erythrocyte volume/total erythrocyte volume) increases with increasing splenic weight, apparently independently of the underlying haematological disorder, the splenic erythrocyte content varying only with body haematocrit. Hyperhaemolysis is a regular finding in enlarged spleens. The splenic erythrocyte sequestration rate increases with increasing splenic erythrocyte pool and is much higher in myeloproliferative than in lymphoproliferative disorders. The total plasma volume is expanded in splenomegalic patients, but the splenic plasma volume constitutes only a minor part of this increment. The anaemia of splenomegalic patients is a consequence of splenic erythrocyte concentration and hypersequestration combined with expansion of the plasma volume. Frequently the total erythropoietic capacity is reduced. Splenectomy is regularly followed by an increased venous haemoglobin concentration.

To determine the fate and distribution of chromium during lactation, six lactating women (25-38 y old) were given three doses of the tracer 53Cr (7.55 micromol/d, or 400 microg/d) on days 1, 2, and 3 of the study. Diet records, blood samples taken while subjects were fasting, and 24-h composite milk and urine samples were collected from day 0 to day 6. Fasting blood samples, morning milk samples, and 24-h urine samples were also collected on days 8, 10, 15, 30, 60, and 90. 53Cr and natural and total chromium concentrations in biological fluids were measured with gas chromatography-mass spectrometry and total urinary chromium was measured with atomic-absorption spectrometry. 53Cr was detectable in serum 2 h after dosing and continued to be detected from day 30 to day 60. Changes in total serum chromium concentration in response to the oral dose suggested that chromium concentrations in blood were not tightly regulated. 53Cr was not detected in breast milk and no significant changes in natural chromium concentration in milk were observed in response to the oral doses, suggesting that breast-milk chromium concentrations are independent of intake. The estimated chromium intake of exclusively breast-fed infants was 2.5 nmol/d (0.13 microg/d), below the lower end of the range of estimated safe and adequate daily dietary intakes (10-40 microg/d) for infants 0-6 mo of age. The baseline chromium concentration in urine and the minimum 53Cr absorption in lactating women were comparable with values for nonpregnant, nonlactating subjects. Chromium losses in breast milk do not appear to be compensated for via increased absorption or decreased excretion.

There has been a resurgence of interest in the use of metal on metal bearings in total hip arthroplasty. Although the use of metal on metal bearing couples would eliminate or substantially reduce particulate polyethylene generation (depending on the presence or absence of polyethylene in the implant system), there is concern about the potential for increased particulate and ionic metal generation in comparison with polyethylene on metal bearings. These metallic degradation products may be transported away from the implant site and distributed systemically. Chromium concentrations in the serum and urine and cobalt concentrations in the serum were measured in subjects with cobalt chromium alloy metal on metal total hip replacements and in controls without implants. Eight subjects with long term (> 20 years) McKee-Farrar total hip replacements had 9-fold elevations in serum chromium, 35-fold elevations in urine chromium, and at least 3-fold elevations in serum cobalt concentrations in comparison with controls. Six subjects with short term (< 2 years) metal on metal surface replacement arthroplasties had 3-fold elevations in serum chromium, 4-fold elevations in urine chromium, and 4-fold elevations in serum cobalt concentrations in comparison with subjects with McKee-Farrar implants. Although the toxicologic importance of these trace metal elevations has not been established, serum and urine metal concentrations may be useful markers for the tribologic performance of metal on metal bearings.

One new approach to cancer therapy is based on the adoptive transfer of tumor-specific cytotoxic T cells and anti-CD25 antibodies. In the present study, CD8+ and IFN-gamma secreting T lymphocytes (CTLs) were enriched as tumor-specific cytotoxic T cells from spleen lymphocytes of mice bearing the Renca tumor (a murine renal carcinoma line originating from a BALB/c mouse) after stimulation with tumor cells. An anti-CD25 IL-2Ralpha(anti-CD25) mAb from hybridoma PC61 was used for depletion for CD4(+)CD25(+) regulatory T (Treg) cells. Treatment-efficacy for tumor-bearing mice was compared using 4 systems: 1, whole spleen lymphocytes stimulated with tumor cells in vitro from tumor-bearing mice; 2, CTLs; 3, anti-CD25 mAbs; 4, CTLs and anti-CD25 mAbs. At the 50th day after tumor inoculation, in the group which received anti-CD25 mAb for depletion of T cells and inoculation of CTLs, tumors had disappeared and no re-growth was observed. In contrast, all mice of the non-treated and other three groups, treated with whole spleen cells alone, CTLs alone and anti-CD25 mAb alone, had died. These results showed that a combination of Treg cell-depletion using anti-CD25 mAbs and CTL administration is a feasible approach for treatment of cancers which warrants further exploration in the clinical setting.

Irritable bowel syndrome (IBS) is a heterogeneous condition and defined according to symptoms. Low-grade inflammation has been associated with IBS, particularly that following infection, but whether altered intestinal permeability profiles relate to irritable bowel subtype or onset is uncertain. Our aim was to compare small and large intestinal permeability in various subtypes of IBS to healthy controls.Intestinal permeability was measured using 1.8 MBq of 51Cr-EDTA and collecting urine over 24 h; Study 1: patients with diarrhea-predominant postinfectious IBS (N=15), constipation-predominant IBS (N=15), and healthy controls (N=15); Study 2: two groups of diarrhea-predominant IBS (D-IBS), one with a history of onset after acute gastroenteritis (postinfectious) (N=15) and the other without such a history (nonpostinfectious) (N=15) both compared with healthy controls (N=12).Permeability expressed as percentage of total dose excreted in urine (median [inter-quartile range]). Study 1: Proximal small intestinal permeability was increased in postinfectious IBS (0.19 [0.12-0.23]) in contrast to constipated IBS (0.085 [0.043-0.13]) and controls (0.07 [0.035-0.19]) (p=0.02). IBS patients with eczema, asthma, or hayfever had increased proximal small intestinal permeability compared with IBS patients without atopy (p=0.02). Study 2: Small intestinal permeability was greater in nonpostinfectious diarrhea-predominant IBS (0.84 [0.69-1.49]) compared with postinfectious IBS (0.43 [0.29-0.63], p=0.028) or controls (0.27 [0.2-0.39]), p=0.001).Small intestinal permeability is frequently abnormal in diarrhea-predominant IBS. Those without a history of infectious onset appear to have a more severe defect.

Analysis of perforin-deficient mice has identified the cytolytic pathway and perforin as the preeminent effector molecule in T cell-mediated control of virus infections. In this paper, we show that mice lacking both granzyme A (gzmA) and granzyme B (gzmB), which are, beside perforin, key constituents of cytolytic vesicles, are as incapable as are perforin-deficient mice of controlling primary infections by the natural mouse pathogen ectromelia, a poxvirus. Death of gzmA×gzmB double knockout mice occurred in a dose-dependent manner, despite the expression of functionally active perforin and the absence of an intrinsic defect to generate splenic cytolytic T cells. These results establish that both gzmA and gzmB are indispensable effector molecules acting in concert with perforin in granule exocytosis-mediated host defense against natural viral pathogens.

To validate and improve a method of red cell volume determination by use of a stable isotope of chromium.Twelve subjects were sequentially injected with red blood cells labeled with a stable isotope of chromium (53Cr) and red blood cells labeled with radioisotopic chromium (51Cr). Measurement of 53Cr dilution was by gas chromatography/mass spectrometry. Measurement of 51Cr dilution was by gamma counter.Comparison of the two methods led to results that differed on average by 34.5 +/- 45.0 mL (1.8 +/- 2.2%), 0.3 to 3.2%, 95% confidence interval.Measurement of red cell volume by use of a stable isotope of chromium is accurate, with potential applications including measurement in pregnant women and children or other groups in whom exposure to radioisotopes is undesirable.

Erythrocyte volumes of thoroughbred horses were measured. The volumes of splenectomized horses and sham-operated horses 2 hr after injection of 50Cr-tagged erythrocytes (at rest) and during maximal exercise were measured using the non-radioactive isotope 50Cr. Because splenic erythrocytes are released into circulation during exercise, it was estimated that the erythrocyte volumes of the sham-operated horses during maximal exercise are larger than those of the horses at rest. However, the erythrocyte volumes of the sham-operated horses at rest were about equal to those during maximal exercise. In the splenectomized horses, furthermore, erythrocyte volumes at rest and those at exercise were nearly equal. From these results, blood stored in the equine spleen is gradually mixed with circulating blood, and it was clarified that the phenomenon was completed within 2 hr. Although it is basically impossible to measure the circulating erythrocyte volume at rest using the erythrocyte tagged method, we observed that it is possible to measure the total erythrocyte volume using the 50Cr method. Also, the plasma volumes of the splenectomized horses during maximal exercise were found to be slightly smaller than those at rest. On the other hand, in the sham-operated horses, the plasma was decreased by a large quantity after maximal exercise. Therefore, it was suggested that the spleen participates in the phenomenon involving the disappearance of plasma from circulation due to exercise.

We examined utilization of selenomethionine (SeMet) and selenite in six lactating (L) and six nonlactating (NL) women, 2-3 mo postpartum, and seven never-pregnant (NP) women by use of stable-isotope tracers. All groups had similar selenium status at the start of the study. Significantly more selenium from SeMet than from selenite was absorbed and appeared in plasma in all groups. Milk contained more selenium from apparently absorbed SeMet than from selenite. More selenium from apparently absorbed selenite than from SeMet appeared in urine of NP and NL subjects whereas L subjects had approximately the same amount of selenium from apparently absorbed selenite and SeMet in their urine. All groups retained significantly more selenium from SeMet than from selenite; L women retained more selenium from selenite than did the other two groups. Absorption and retention of selenium from SeMet in L women did not appear to be significantly different from that in other women, suggesting that selenium requirements during lactation are increased mainly because of milk losses.

Specific lymphocyte-mediated cytotoxicity to cytomegalovirus (CMV) in eight infants (six to 27 months old) with congenital CMV infection and in the mothers of six of these infants was evaluated with use of a 51chromium (51Cr)-release microassay. The control population consisted of 25 normal newborns, children, and adults. The titers of indirect hemagglutinating (IHA) antibody to CMV in the infected infants ranged from 1:16 to 1:1,024. All of these infants had detectable specific immune release of 51Cr that ranged from 3.3% to 48.9% (mean +/-SE, 21.0%+/-5.6%). The mothers of these infants demonstrated significantly elevated titers of IHA antibody to CMV (geometric mean titer, 1:410) as compared with a mean titer of 1:22 in controls (t = 5.71; P less than 0.001) but showed significantly depressed specific immune release (9.2% +/- 3.2%) compared with that of normal seropositive controls (24.8% +/- 2.8%; t = 3.31; P less than 0.001). In addition, two adult nulliparous women with persistent CMV viruria were also found to have depressed specific immune release to CMV (10.8% and 16.2%). These data suggest that a specific impairment in cell-mediated immunity to CMV occurs in mothers of infants with congenital CMV infection and in some persons who persistently excrete CMV.

Hemodynamic data were obtained in 13 cirrhotic patients with severe portal hypertension, undergoing combined hepatic vein, umbilicoportal vein, and superior mesenteric artery catheterization. The relative clearance of indocyanine green, the portohepatic gradient (difference between the free portal venous pressure and the free hepatic venous pressure), and the estimated hepatic blood flow were measured. The portal fraction (PF) of total hepatic blood flow was calculated in all patients using indicator dilution curves obtained from the portal bifurcation, a right hepatic vein, and when possible a left hepatic vein (six cases) after injection of 51Cr-labeled red blood cells (51Cr RBC) into the superior mesenteric artery. Flows were overestimated because of loss of indicator through spontaneous portosystemic shunts; however, the ratio between hepatic and portal indicator dilution curves can be used to calculate the portal fraction of total hepatic blood flow since no extrahepatic shunts existed after the bifurcation of the portal vein (as shown on portography). In 10 patients, 15 series of curves were calculable and the PF varied between 30.1 and 100% (mean ± SE: 71.1 ± 6.2%). In the three other patients, only delayed activity from recirculation was detected from portal and hepatic vein samples and PF was 0%; in these three cases, portography and arteriography revealed spontaneous portacaval shunting with reverse and/or stagnant circulation in the portal vein. In the 13 patients, no correlation existed between PF and the relative clearance of indocyanine green or the portohepatic gradient, parameters generally used as indices of severity in cirrhosis. In 10 patients, no correlation was found between PF and the estimated hepatic blood flow. These data indicate that 51Cr RBC dilution curves can be used for the estimation of the portal fraction of total hepatic blood flow in conscious cirrhotic patients before portacaval shunts. Using this methodology, it could be assessed whether any critical level of portal fraction exists above which poor clinical results occur after portacaval shunting. This measurement could eventually be helpful in determining the appropriate surgical procedure to be applied in individual cases. ImagesFIG. 4FIG. 5aFIG. 5bFIG. 5c

The ability of guinea pigs to make immune responses to GA, a linear random copolymer of L-glutamic acid and L-alanine, GT, a random linear copolymer of L-glutamic acid and L-tyrosine, and PLL, a linear homopolymer of L-lysine, is controlled by different autosomal dominant genes specific for each of those polymers. We have investigated the relationship between the PLL gene and the GA and GT immune response genes by simultaneously immunizing random-bred Hartley strain guinea pigs with GA and PLL, GT and PLL, or GA and GT. In most Hartley guinea pigs the ability to respond immunologically to GA and to PLL is inherited together; that is, most animals responding to GA respond to PLL and vice versa. However, a few animals respond to either GA or to PLL but not both, demonstrating that the GA and PLL immune response genes are not identical but linked in most Hartley animals. Conversely, when simultaneously immunized with GT and PLL, most Hartley guinea pigs respond to either PLL or GT but not both, indicating that GT and PLL responsiveness tends to segregate away from each other. Thus, the GT and PLL immune response genes also are not inherited independently but, rather, behave as alleles or pseudoalleles. Similar results are observed when Hartley guinea pigs are simultaneously immunized with GA and GT. The ability to respond to GA segregates away from the ability to respond to GT. Our studies demonstrated that the specific immune response genes thus far identified in guinea pigs controlling the ability to respond to GA, GT, and PLL, respectively, are found on the same chromosome. In most Hartley animals, the GA and PLL immune response genes are often linked, i.e. occur on the same chromosome strand, and tend to behave as alleles or pseudoalleles to the GT immune response gene.

Summary. The exchangeable splenic platelet pool (ESPP) was studied with epinephrine infusion and platelet labelling with 51Cr in five healthy students, 10 patients with idiopathic thrombocytopenic purpura (ITP) and 10 patients with splenomegaly. Five of the ITP-patients were studied after splenectomy. Platelet recovery of infused labelled platelets was calculated in all subjects and also in nine healthy volunteers who had been splenectomized for traumatic rupture of the spleen. Spleen size was determined by gamma camera scintigraphy. It was shown that the spleen is the only site of an exchangeable platelet pool in ITP and that this pool was of the same size in ITP-patients as in the normal controls, viz ∼30% of the total body platelet mass. In patients with splenomegaly the ESPP was larger than that in controls and ITP-patients. A highly significant correlation was found between the ESPP and the spleen volume. In splenectomized and in non-splenectomized ITP-patients platelet recovery was significantly less than in their respective control groups, indicating that a proportion of the labelled platelets was immediately removed from the circulation after infusion into an ITP recipient and that the recovery of labelled platelets cannot be used as a measure of the ESPP in ITP. It is suggested that the early destruction of platelets may be due to slight damage to the platelets during the labelling procedure. These damaged platelets can survive in a normal recipient, but are destroyed when infused into the‘milieu’ of an ITP-patient.

The cells in the mouse thymus which respond to PHA and have the distribution characteristics of recirculating lymphocytes upon infusion into isologous recipients are markedly enriched 2 days after treatment of mice with proper doses of cortisone acetate. However, a similar increase is not observed in lymph nodes and spleen. During regeneration of the steroidinvoluted thymus, the proportion of recirculating thymic cells disappeared more rapidly than the relative PHA-responsiveness of the cell population. These findings indicate that all PHA-responsive thymocytes may not have recirculating capacity. The recirculating cells present in the thymus and lymph nodes have no tendency to distribute to the thymus, but tend to accumulate in lymph nodes. This fact, together with the finding that in vitro trypsin treatment of the cells temporarily abolishes their lymph node-seeking capacity, are taken as evidence that the recirculating cells in the thymus have undergone their maturation within this organ whereby they have gained membrane receptors which determine their distribution characteristics.

SummaryIn an attempt to define the mechanism by which the reticuloendothelial stimulant, glucan, profoundly enhances susceptibility and conversely that the RES depressant, methyl palmitate, produces a resistance to intravenously administered endotoxins, the vascular clearance and tissue distribution of intravenously administered 51Crlabeled S. enteritidis endotoxin were studied. The vascular clearance of the labeled endotoxin preparation was biphasic consisting of an early, or fast, component followed by a slow component. The RES stimulant, glucan, enhanced both phases of removal and produced a significant increase in the hepatic localization of the endotoxin. Splenic and lung uptake were unaltered. In contrast, methyl palmitate exerted no influence on the vascular removal rate of endotoxin, but induced a significant decrease in splenic uptake. The administration of heparin, which has been previously demonstrated to modify endotoxin lethality, decreased the early phase of vascular clearance, but did not m...

Two siblings with the rare phenotype Rhnull are described; the parents are related to each other. Serologic investigation of the family gives indirect support for the action of an amorphous gene. As demonstrated in the propositus, the splenectomy resulted in health improvement. Titration tests in 1,803 unrelated blood donors revealed 4 heterozygous — types. On the basis of this calculation one can assume one Rhnull patient among 6 million inhabitants.

Dilutional Anemia of Splenomegaly: An Indication for Splenectomy Charles Hess;Carlos Ayers;Richard Wetzel;Daniel Mohler;William Sandusky; Annals of Surgery

Effect of Estradiol-17, β on the Magnitude and Distribution of Uterine Blood Flow in Nonpregnant, Oophorectomized Ewes

SUMMARY The pathogenicity of Fasciola hepatica was studied in a group of six sheep experimentally infected seven weeks after mating. Eosinophilia and raised levels of gamma globulin were early changes in the blood, followed later by the onset of a progressive anaemia accompanied by loss of liveweight. At eight weeks after infection, voluntary food intake and dry matter retention were significantly (P

SUMMARY Red cell survival studies using 51 Cr-labelled erythrocytes were carried out on 14 subjects with protein malnutrition (12 kwashiorkor and two marasmus). The T½ varied from 3 to 20 days with a mean of 12.4 days. There was significant accumulation of radioactivity demonstrable on scanning over the liver and spleen in three kwashiorkor patients. Erythrocytes of three kwashiorkor patients showed shortened T½ both when auto-transfused and when injected into three normal controls. The T½ of erythrocytes from two normal donors injected into four subjects with kwashiorkor was also considerably reduced. After protein repletion, auto-transfusion in nine cases showed a T½ which varied from 20 to 29 days with a mean of 23.7 days. This investigation has shown considerably reduced erythrocyte survival in patients with protein malnutrition (kwashiorkor and marasmus); this shortened survival time appears to be due to both corpuscular and extra-corpuscular factors. Following protein feeding considerable improvement in the erythrocyte survival took place. Since improvement in erythrocyte survival occurred on a low iron-containing protein diet without haematinics or vitamin supplements it is considered that protein depletion is principally responsible for this shortened survival.

BILIRUBIN metabolism and excretion in children with biliary atresia is complex and poorly understood. Jaundice may not be apparent until the infant is three or four weeks of age. Rarely, as in a case reported by Sterling,1 the jaundice disappears, and for weeks the serum bilirubin levels are normal. Usually, however, the serum bilirubin levels continue to rise for several weeks after birth and thereafter remain relatively stable. Like the normal infant, the child with biliary atresia is continuously producing bile pigment from the breakdown of hemoglobin. Since the serum bilirubin levels do not continue to rise one must assume . . .

Mean blood volume in 13 patients with acute intermittent porphyria was 80 per cent of normal (p less than 0.01). There was a parallel reduction in both plasma and total red-cell volumes. In seven patients the half-life of 51Cr-labeled red cells ranged from 24 to 32 days. The finding of a reduced red-cell volume without accelerated red-cell destruction indicated that effective erythropoiesis was reduced. Ferrokinetic studies in one patient also demonstrated decreased erythropoiesis (plasma iron turnover was 70 per cent of normal). The bilirubin production rate in six patients was proportional to the circulating red-cell mass (r = 0.84), indicating that there was no increase in the early labeled fraction of bilirubin. In one patient a normal estimate for this fraction was found with the use of glycine-2–14C. These results indicate that ineffective erythropoiesis is not increased in acute intermittent porphyria and also suggest that there is no marked increase in hepatic bilirubin production.

The red cell mass, splenic red cell pool, and plasma volume have been measured in a series of 64 patients with splenomegaly due to haematological disorders. In 45 patients the total blood volume was increased, this being due to expansion of the plasma volume which was closely correlated with spleen size. There was no significant relationship between the red cell mass and spleen size but the splenic red cell pool increased with increasing spleen size. In 12 patients serial estimations were made whilst changes in spleen size occurred as a result of therapy or progression of the disease. The red cell mass did not alter but the splenic red cell pool showed a significant change from 8.9 +/- 6.1% of the red cell mass at minimum spleen size to 17.5 +/- 10.5% at maximum spleen size. There was progressive expansion of the plasma volume from 42.9 +/- 5.0 ml/kg at minimum spleen size to 57.4 +/- 15.3 ml/kg at maximum spleen size.

Glomerular filtration rate and renal plasma flow were simultaneously determined in comparable groups of 43 diabetics less than 40 years of age and with a duration of diabetes less than 10 years and 32 control subjects. The average glomerular filtration rate in the diabetic group was significantly higher than that in the control group (P0.01). The average renal plasma flow in the diabetic group was found to be significantly lower than that in the control group (P0.05). The filtration fraction in both male and female diabetics was significantly higher than in the male and female control groups (P0.001). These changes were found to be present even in recent juvenile diabetics with disease of a duration of less than one year. No correlation was apparent between the average levels of serum growth hormone and glomerular filtration rate.The urinary protein excretion was determined in 36 diabetic and 38 healthy subjects comparable with regard to glomerular filtration rate. In the diabetic group there was a greater frequency of cases with higher protein excretion rates (P0.02). The average protein excretion rate was increased even in diabetics with less than one year's duration of the disease.The results of the changes in renal haemodynamics in subjects with recent and short-term diabetes are compatible with the presence of a constrictive state of the vas efferens leading to an increase in the filtration pressure. The increase in protein excretion rate may similarly be a consequence of this process or of an increase in the glomerular permeability with augmented molecular sieving of proteins or both.

(1966). Direct Intravenous Injection of 51Chromic Chloride Compared with 125I-Polyvinylpyrrolidone and 131I-Albumin in the Detection of Gastrotestinal Protein Loss. Scandinavian Journal of Clinical and Laboratory Investigation: Vol. 18, No. 4, pp. 405-416.

Muscle capillary permeability to 51Cr-EDTA was studied in the exercising forearm of 17 normal human adults. — The permeability data were obtained by means of the indicator dilution technique and the fractional unidirectional transcapillary loss or extraction, E, of 51Cr-EDTA was calculated using five conceptually different equations. They all yielded practically the same E value. This supports the contention that the indicator dilution technique can give a valid measure of the permeability characteristics of the skeletal muscle capillary wall. These permeability characteristics are expressed as the capillary diffusion capacity, CDC, which is defined as the maximal unidirectional transcapillary flux of a permeable tracer per unit concentration difference per 100 g skeletal muscle. In normal man CDCCr-EDTA reaches its maximum value of 3.95, SD ± 0.50 moles/100 g·min per mole/ml at plasma flow levels exceeding 15 ml/100g·min.

In the preceding paper (Lockner 1966 b) a new method for the quantitative estimation of erythropoiesis is introduced. This method is based on the principle that the fraction of plasma iron transported per day (PIT) and incorporated into the circulating erythrocytes (RBCIT) becomes a quantitative measure of erythropoiesis, if recircula-tion of radioiron can be inhibited. This was achieved by intravenous injections of inactive iron. It is the purpose of this paper to report the application of this method to cancer patients. A preliminary account of this work has been presented (Lockner 1964).

Cellular cytotoxicity reactions to human leukemia cells were measured with a sensitive, quantitative assay of cell-mediated immunity. Freshly explanted leukemia cells, rather than tissue culture cells, were tested to ensure that the antigens detected were not acquired in vitro. Identical twins, one member of each pair having leukemia, were used to minimize genetic variables and histocompatibility differences, thereby providing comparable normal cells to control for the leukemic cells. Leukemia-associated antigens were detected on the cells of seven of ten leukemic patients. In no instance was reactivity observed against only the cells of the normal twin. Lymphocytes from three identical twins, seven parents, two siblings and eight normal unrelated individuals were cytotoxic for cells from the leukemic patients but not for cells from these patients' normal identical twins. Lymphocytes from adults had a much higher incidence of reactivity (46%) against the leukemic cells than did the cells of children (9%). Positive lymphocyte cytotoxicity reactions to leukemia-associated antigens indicate previous sensitization which could have resulted from infection with an environmental agent such as a virus.

The urinary excretion of 51Cr was studied in conjunction with studies of erythrocyte survival, fecal blood loss, and erythrocyte volume in 54 patients with various hematologic disorders. It was confirmed that excretion of 51Cr is predominantly urinary, and the rate approximates a simple exponential function. In anemias unaccompanied by hemolysis or blood loss, the halftime for urinary excretion of erythrocyte-bound 51Cr (Tu½) ranged from 43 to 63 days. Tu½ was shortened in patients with hemolytic disease but prolonged in patients with severe blood loss; these deviations were proportional to the severity of hemolysis or blood loss, respectively. Determination of the rate of urinary 51Cr excretion may prove to be useful in differentiating various causes of anemia, especially because (1) it distinguishes severe or chronic blood loss from hemolysis, which is not possible with conventional erythrocyte survival studies; (2) the need for multiple venipunctures may be obviated; (3) fluctuations in blood volume or erythrocyte volume do not appear to affect the rate of urinary 51Cr excretion; (4) only a very small dose of radiochromium is needed.

The aim of the present work was to study to what extent the vascular area of the leg contributes to the body hematocrit phenomenon, i.e. the fact that the total body hematocrit (hctbody) is about 10% lower than the large vessel hematocrit (hctl.v.).The hematocrit of the leg (hctleg) was calculated from the mean circulation times of plasma and red cells through the vascular bed of the lower extremity. These values were determined at rest and during exercise by the arteriovenous equilibration technique after injection of 125I-labelled albumin and 51Cr-labelled red cells into an antecubital vein and sampling from a peripheral artery and the femoral vein.In 14 normal resting subjects the average hctleg was 90.2% and hctbody 86.9% of hctl.v.. Hctleg was significantly higher than hctbody. During exercise hctleg was also found to be significantly lower than hctl.v. and higher than hctbody, but the ratio between hctleg and hctl.v. was not significantly lower during exercise than during rest.It is concluded that t...

SummaryStudies in three hypophysectomized monkeys revealed markedly reduced red cell volume, plasma volume, percentage of nucleated erythron in the bone marrow, and plasma erythropoietin activity. Corticosteroids and thyroxine, given alone or in combination, had no significant effect on these abnormalities; the combined growth hormone and thyroxine therapy, however, restored all the abnormalities to normal.

In late toxemia of pregnancy the volume of the whole blood and the absolute amount of the serum proteins decreases not only in the developed clinical picture of illness, but also even before any clinical symptoms occur. In these potential patients the volume of the whole blood was lower by about 495 ml. and the absolute amount of the serum proteins about 23.60 Gm. than in physiologic pregnancy. These findings at the beginning of late toxemia of pregnancy may serve as an “early indicator” and may also help in the prevention of this disease.