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Chromosomal rearrangements of the human KMT2A/MLLgene are associated with de novoas well as therapy-induced infant, pediatric, and adult acute leukemias. Here, we present the data obtained from 3401 acute leukemia patients that have been analyzed between 2003 and 2022. Genomic breakpoints within the KMT2Agene and the involved translocation partner genes (TPGs) and KMT2A-partial tandem duplications (PTDs) were determined. Including the published data from the literature, a total of 107 in-frame KMT2Agene fusions have been identified so far. Further 16 rearrangements were out-of-frame fusions, 18 patients had no partner gene fused to 5’-KMT2A, two patients had a 5’-KMT2Adeletion, and one ETV6::RUNX1patient had an KMT2Ainsertion at the breakpoint. The seven most frequent TPGs and PTDs account for more than 90% of all recombinations of the KMT2A, 37 occur recurrently and 63 were identified so far only once. This study provides a comprehensive analysis of the KMT2Arecombinome in acute leukemia patients. Besides the scientific gain of information, genomic breakpoint sequences of these patients were used to monitor minimal residual disease (MRD). Thus, this work may be directly translated from the bench to the bedside of patients and meet the clinical needs to improve patient survival.

PCR of TCR/Ig gene rearrangements is considered the method of choice for minimal residual disease (MRD) quantification in BCP-ALL, but flow cytometry analysis of leukemia-associated immunophenotypes (FCM-MRD) is faster and biologically more informative. FCM-MRD performed in 18 laboratories across seven countries was used for risk stratification of 1487 patients with BCP-ALL enrolled in the NOPHO ALL2008 protocol. When no informative FCM-marker was available, risk stratification was based on real-time quantitative PCR. An informative FCM-marker was found in 96.2% and only two patients (0.14%) had non-informative FCM and non-informative PCR-markers. The overall 5-year event-free survival was 86.1% with a cumulative incidence of relapse (CIR5y) of 9.5%. FCM-MRD levels on days 15 (HzR 4.0, p< 0.0001), 29 (HzR 2.7, p< 0.0001), and 79 (HzR 3.5, p< 0.0001) associated with hazard of relapse adjusted for age, cytogenetics, and WBC. The early (day 15) response associated with CIR5yadjusted for day 29 FCM-MRD, with higher levels in adults (median 2.4 × 10−2versus 5.2 × 10−3, p< 0.0001). Undetectable FCM- and/or PCR-MRD on day 29 identified patients with a very good outcome (CIR5y= 3.2%). For patients who did not undergo transplantation, day 79 FCM-MRD > 10−4associated with a CIR5y= 22.1%. In conclusion, FCM-MRD performed in a multicenter setting is a clinically useful method for MRD-based treatment stratification in BCP-ALL.

Minimal residual disease (MRD) measured by PCR of clonal IgH/TCR rearrangements predicts relapse in T-cell acute lymphoblastic leukemia (T-ALL) and serves as risk stratification tool. Since 10% of patients have no suitable PCR-marker, we evaluated flowcytometry (FCM)-based MRD for risk stratification. We included 274 T-ALL patients treated in the NOPHO-ALL2008 protocol. MRD was measured by six-color FCM and real-time quantitative PCR. Day 29 PCR-MRD (cut-off 10−3) was used for risk stratification. At diagnosis, 93% had an FCM-marker for MRD monitoring, 84% a PCR-marker, and 99.3% (272/274) had a marker when combining the two. Adjusted for age and WBC, the hazard ratio for relapse was 3.55 (95% CI 1.4–9.0, p= 0.008) for day 29 FCM-MRD ≥ 10−3and 5.6 (95% CI 2.0–16, p= 0.001) for PCR-MRD ≥ 10−3compared with MRD < 10−3. Patients stratified to intermediate-risk therapy on day 29 with MRD 10−4–<10−3had a 5-year event-free survival similar to intermediate-risk patients with MRD < 10−4or undetectable, regardless of method for monitoring. Patients with day 15 FCM-MRD < 10−4had a cumulative incidence of relapse of 2.3% (95% CI 0–6.8, n= 59). Thus, FCM-MRD allows early identification of patients eligible for reduced intensity therapy, but this needs further studies. In conclusion, FCM-MRD provides reliable risk prediction for T-ALL and can be used for stratification when no PCR-marker is available.

Adults with acute lymphoblastic leukemia (ALL) do worse than children. From 7/2008 to 12/2014, Nordic and Baltic centers treated 1509 consecutive patients aged 1–45 years with Philadelphia chromosome-negative ALL according to the NOPHO ALL2008 without cranial irradiation. Overall, 1022 patients were of age 1–9 years (A), 266 were 10–17 years (B) and 221 were 18–45 years (C). Sixteen patients (three adults) died during induction. All others achieved remission after induction or 1–3 intensive blocks. Subsequently, 45 patients (12 adults) died, 122 patients relapsed (32 adults) with a median time to relapse of 1.6 years and 13 (no adult) developed a second malignancy. Median follow-up time was 4.6 years. Among the three age groups, older patients more often had higher risk ALL due to T-ALL (32%/25%/9%, P<0.001), KMT2A rearrangements (6%/5%/3%, P<0.001) and higher day 29 residual leukemia for B-lineage (P<0.001), but not T-ALL (P=0.53). Event-free survival rates (pEFS5y) were 89±1% (A), 80±3% (B) and 74±4% (C) with significant differences only for non-high risk groups. Except for thrombosis, pancreatitis and osteonecrosis, the risk of 19 specified toxicities was not enhanced by age above 10 years. In conclusion, a pediatric-based protocol is tolerable and effective for young adults, despite their increased frequency of higher risk features.

Chromosomal rearrangements of the human MLL/KMT2A gene are associated with infant, pediatric, adult and therapy-induced acute leukemias. Here we present the data obtained from 2345 acute leukemia patients. Genomic breakpoints within the MLL gene and the involved translocation partner genes (TPGs) were determined and 11 novel TPGs were identified. Thus, a total of 135 different MLL rearrangements have been identified so far, of which 94 TPGs are now characterized at the molecular level. In all, 35 out of these 94 TPGs occur recurrently, but only 9 specific gene fusions account for more than 90% of all illegitimate recombinations of the MLL gene. We observed an age-dependent breakpoint shift with breakpoints localizing within MLL intron 11 associated with acute lymphoblastic leukemia and younger patients, while breakpoints in MLL intron 9 predominate in AML or older patients. The molecular characterization of MLL breakpoints suggests different etiologies in the different age groups and allows the correlation of functional domains of the MLL gene with clinical outcome. This study provides a comprehensive analysis of the MLL recombinome in acute leukemia and demonstrates that the establishment of patient-specific chromosomal fusion sites allows the design of specific PCR primers for minimal residual disease analyses for all patients.

An envelope is introduced by using the Hilbert transform to define a complex conjugate to the excitation and response processes of a linear structure. Timelimited stationary excitation is treated in detail, and the complex correlation function is shown to follow from its stationary equivalent by use of a suitable differential operator. Simple expressions are derived for the case of rational spectral density, and a parametric study of the influence of the frequency content is carried out. It is found that envelope crossings can be predicted by use of stationary measures for the mean crossing frequency and the bandwidth of the response combined with the nonstationary intensity. This is computationally important as these parameters are often available in closed form. Finally, the envelope is used to study the firstpassage probability.

Clipped normal processes are considered from the point of view of modeling transient loads on structures. Sample curves of these types of processes are pulse like, each pulse corresponding to an exceedance Of the zero level by a Gaussian process. The pulses are the more narrow the larger negative is the mean of the Gaussian process. In contrast to current transient load models the pulses are of random shape. How to calculate bounds on the mean upcrossing rate of any level by a sum of several clipped normal processes is demonstrated. For high levels these bounds are close. Thus, they may be used to obtain an accurate evaluation of a wellknown upper bound on the probability of the sum exceeding the level within a given time period. The particular example of the sum of several mutually independent, stationary clipped normal processes permits an exact calculation of the upcrossing rate of any constant level. The upcrossing problem is, in fact, solved by observing the equivalence of the problem with a Gaussian vector process outcrossing problem from a certain convex polyhedral region in the ndimensional space. Reliability theoretical methods for this case have been reported earlier. These methods also make it possible to calculate a lower bound on the exceedance probability giving an evaluation of the upper bound as an approximation to the exact probability.

The extremevalue distribution for the von Mises stress is determined. The stress components are stationary Gaussian processes. An upcrossing of a stress level by the von Mises stress is described as the outcrossing of an ellipsoid by the stress component vector process. The extremevalue distribution for the von Mises stress is expressed in terms of the mean outcrossing rate of the ellipsoid. The mean outcrossing rate is determined by Rice's formula. Code formats for nonlinear stress combinations are compared for an example.

A small group of children with acute lymphoblastic leukemia (ALL) have a preleukemic phase of pancytopenia followed by a period of spontaneous remission before the diagnosis (pre-ALL). A 6-year-old girl presented with pancytopenia, fever, and myelodysplasia. Following transient remission pre-B ALL was diagnosed 14 months later. Clonal B-lineage blasts at the period of pancytopenia were identified retrospectively. The interval between pre-ALL and ALL-diagnosis was longer than previously reported. The infection was clinically severe and might have induced a significant endogenous corticosteroids production resulting in the long-lasting remission. The case supports the adrenal and the Coley’s toxin hypothesis in leukemogenesis.

Recently we found that the incidence of restenosis after carotid endarterectomy was significantly higher in patients homozygous for the normal genotype of mannose-binding lectin (MBL2) than in with patients with MBL2variant genotypes. Several growth factors are also known to contribute to restenosis. Therefore, we investigated whether early postoperative changes in serum vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and platelet-derived growth factor (PDGF) concentrations and MBL2genotypes interact in the development of restenosis.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

The TEL-AML1translocation, t(12;21)(p13;q22), is one of the most frequent genetic aberrations in childhood B-cell precursor acute lymphoblastic leukemia (ALL), where it occurs in 25 of all cases. In contrast, the translocation is seen in only 3 of adult ALL cases. Evidence suggests that the TEL-AML1translocation occurs in utero in 1 of all newborn children at cell levels of 10−3to 10−4. In this study, we explore the prevalence of TEL-AML1–positive cells in 2 cohorts of healthy blood donors by real-time and nested reverse transcription-polymerase chain reaction. Overall, TEL-AML1–positive cells were demonstrated in 10 of 2005 healthy donors, that is, a prevalence of 0.5 (95 confidence interval, 0.2-0.3). The level of TEL-AML1–positive cells was estimated to 10−5to 10−6. The observed prevalence of TEL-AML1–positive cells in healthy adults is of the same order of magnitude as the prevalence reported in healthy newborns, but the observed cell level of 10−5to 10−6is much lower. These data indicates that prenatal TEL-AML1subclones does not persist throughout adult life at cell levels of 10−3to 10−4. The findings are compatible with the risk of t(12;21)(p13;q22) ALL correlating with the total number of TEL-AML1–positive cells in peripheral blood in both childhood and adulthood.

Locked nucleic acid (LNA®) is a modified DNA with increased binding affinity for complementary DNA sequences. Our strategy was to use this property of LNA to inhibit undesired PCR amplification (e.g., from contaminating genomic DNA) in a cDNA-based assay. By placing a short complementary LNA sequence in intronic DNA, the aim was to inhibit the amplification of genomic DNA without affecting the amplification of reverse-transcribed spliced mRNA. LNA was designed to bind within intron 5 in the x-box binding protein 1 (XBP1) gene. An irrelevant LNA oligonucleotide served as a negative control. In both PCR and real-time PCR, the addition of LNA showed blocking of the amplification of genomic XBP1 but not cDNA XBP1. To test the effect of melting temperature (Tm) on the LNA, we investigated the number of LNA nucleotides that could be replaced with DNA nucleotides and still retain the blocking activity. More than three DNA nucleotides reduced the LNA inhibition ability. The sequence specificity of the LNA was tested by investigating the number of LNA nucleotide mismatches permitted. The introduction of one mismatch maintained the inhibition of genomic amplification whereas two mismatches reduced the amplification. Our results show that LNA may be used to enhance the specificity of PCR by eliminating unwanted PCR products.

Variant alleles in the mannose-binding lectin (MBL) gene (mbl2) causing low levels of functional MBL are associated with susceptibility to different infections and are common in areas where malaria is endemic. Therefore, we investigated whether MBL variant alleles in 551 children from Ghana were associated with the occurrence and outcome parameters of Plasmodium falciparum malaria and asked whether MBL may function as an opsonin for P. falciparum. No difference in MBL genotype frequency was observed between infected and noninfected children or between children with cerebral malaria and/or severe malarial anemia and children with uncomplicated malaria. However, patients with complicated malaria who were homozygous for MBL variant alleles had significantly higher parasite counts and lower blood glucose levels than their MBL-competent counterparts. Distinct calcium-dependent binding of MBL to the membrane of P. falciparum-infected erythrocytes, which could be inhibited by mannose, was observed. Further characterization revealed that MBL reacted with a P. falciparum glycoprotein identical to the 78-kDa glucose-regulated stress protein of P. falciparum. MBL seems to be a disease modifier in clinical malaria and to function as an opsonin for erythrocytes invaded by P. falciparum and may thus be involved in sequestration of the parasite, which in turn may explain the association between homozygosity for MBL variant alleles and high parasite counts.

ABSTRACTVariant alleles in the mannose-binding lectin (MBL) gene (mbl2) causing low levels of functional MBL are associated with susceptibility to different infections and are common in areas where malaria is endemic. Therefore, we investigated whether MBL variant alleles in 551 children from Ghana were associated with the occurrence and outcome parameters of Plasmodium falciparummalaria and asked whether MBL may function as an opsonin for P. falciparum. No difference in MBL genotype frequency was observed between infected and noninfected children or between children with cerebral malaria and/or severe malarial anemia and children with uncomplicated malaria. However, patients with complicated malaria who were homozygous for MBL variant alleles had significantly higher parasite counts and lower blood glucose levels than their MBL-competent counterparts. Distinct calcium-dependent binding of MBL to the membrane of P. falciparum-infected erythrocytes, which could be inhibited by mannose, was observed. Further characterization revealed that MBL reacted with a P. falciparumglycoprotein identical to the 78-kDa glucose-regulated stress protein of P. falciparum. MBL seems to be a disease modifier in clinical malaria and to function as an opsonin for erythrocytes invaded by P. falciparumand may thus be involved in sequestration of the parasite, which in turn may explain the association between homozygosity for MBL variant alleles and high parasite counts.

t(12;21)(p1 3;q22), the most frequent chromosomal translocation found in childhood acute lymphoblastic leukemia (ALL), occurs in ~ 25% of B-lineage ALL cases and has been claimed to carry a good prognosis. As part of the Nordic Society of Pediatric Hematology and Oncology (NOPHO) ALL-MRD 95 study, which includes children from Iceland, Norway, and Denmark diagnose d with ALL, patients were screened for the presence of t(12; 21) by reverse transcriptase-polymerase chain reaction (RT-PCR) at diagnosis, and their residual disease was quantified after 4 weeks of induction therapy (prednisolone, vincristine, doxorubicin, i.t. methotrexate) by a competitive, clone-specific, semi-nested PCR analysis. Among 96 children diagnosed with ALL, and quantified for post induction residual disease, 32 were t(12;21)-positive. The median residual disease was similar for B-precursor ALL patients with and without t(12;21) (0.009 vs. 0.03%, P  = 0.12). Al though patients with t(12;21)-positive ALL have been claimed to have a good outcome, these data indicate that this does not reflect a high sensitivity to prednisolone, vincristine, and doxo rubicin given during induction therapy. Med Pediatr Oncol 2003;40:82–87. © 2003 Wiley-Liss, Inc.

The postinduction level of minimal residual disease (MRD) was quantified with a competitive polymerase chain reaction (PCR) technique in 104 children with acute lymphoblastic leukemia (ALL) diagnosed between June 1993 and January 1998 and followed for a median of 4.2 years. A significant correlation was found between the MRD level on day 15 (D15) and day 29 (D29) after the start of induction therapy (rs = 0.70, P < .0001). The 15 patients with T-cell disease had higher D29 MRD than those with B-lineage ALL (P = .01). Age was positively related to D29 MRD (rs = 0.32, P = .001). The 16 patients who had a relapse had higher D15 and D29 MRD levels than the patients who stayed in remission (median levels D15, 1% versus 0.1%,P = .03; D29, 0.4% versus 0.01%,P = .0001). No patients with a MRD level less than 0.01% on D29 have so far had a relapse, whereas the 7-year probability of event-free survival for patients with higher MRD levels was 0.52 (P = .0007). The group of patients with a D29 MRD less than 0.01% included patients with T-cell disease, white blood cell count more than 50 × 109/L at diagnosis, or age 10 years or older, and could not be identified by up-front criteria. The best-fit Cox model to predict the risk of relapse included D29 MRD (P = .004) and age (P = .009). These findings indicate that with the present treatment protocol MRD quantification at an early stage of therapy identifies patients with a very low risk of relapse. Further trials are needed to reveal whether such patients with D29 MRD less than 0.01% can be cured with less intensive chemotherapy, which would reduce the risk of serious late effects as well as the costs of therapy.

The postinduction level of minimal residual disease (MRD) was quantified with a competitive polymerase chain reaction (PCR) technique in 104 children with acute lymphoblastic leukemia (ALL) diagnosed between June 1993 and January 1998 and followed for a median of 4.2 years. A significant correlation was found between the MRD level on day 15 (D15) and day 29 (D29) after the start of induction therapy (rs= 0.70, P< .0001). The 15 patients with T-cell disease had higher D29 MRD than those with B-lineage ALL (P= .01). Age was positively related to D29 MRD (rs= 0.32, P= .001). The 16 patients who had a relapse had higher D15 and D29 MRD levels than the patients who stayed in remission (median levels D15, 1% versus 0.1%,P= .03; D29, 0.4% versus 0.01%,P= .0001). No patients with a MRD level less than 0.01% on D29 have so far had a relapse, whereas the 7-year probability of event-free survival for patients with higher MRD levels was 0.52 (P= .0007). The group of patients with a D29 MRD less than 0.01% included patients with T-cell disease, white blood cell count more than 50 × 109/L at diagnosis, or age 10 years or older, and could not be identified by up-front criteria. The best-fit Cox model to predict the risk of relapse included D29 MRD (P= .004) and age (P= .009). These findings indicate that with the present treatment protocol MRD quantification at an early stage of therapy identifies patients with a very low risk of relapse. Further trials are needed to reveal whether such patients with D29 MRD less than 0.01% can be cured with less intensive chemotherapy, which would reduce the risk of serious late effects as well as the costs of therapy.

OBJECTIVE: To determine whether variant alleles of the mannose binding lectin (MBL) gene causing low serum concentrations of MBL are associated with increased susceptibility to rheumatoid arthritis (RA) and erosive outcome in an inception cohort of patients with early polyarthritis. METHODS: MBL and HLA-DRB1 alleles were determined by polymerase chain reaction in 68 Danish patients with incident early polyarthritis observed for one year. The associations between MBL and specific HLA-DRB1 genotypes and disease outcomes were analyzed. RESULTS: Among the patients with early polyarthritis 7.4% (5/68) and 41.2% (28/68) were homozygous and heterozygous for MBL variant alleles, compared with 2.8% (7/250) and 34.4% (86/250) of healthy controls (p = 0.09), while the corresponding figures in the patients with RA were 10% (5/50) and 42% (21/50) (p = 0.03), and in the patients with erosive RA 18.8% (3/16) and 35.3% (6/16), respectively (p = 0.004). Patients with early polyarthritis homozygous for MBL variant alleles had an increased risk of having erosive RA at inclusion by a factor of 4.7 (p = 0.02) and after one year by a factor of 3.6 (p = 0.04). MBL deficiency was associated with increased levels of C-reactive protein (CRP) and IgM rheumatoid factor (RF) at inclusion (p < 0.05). HLA-DRB1 alleles were not found to be associated with disease outcome. CONCLUSION: MBL variant alleles appear to be weak susceptibility markers for RA, and patients with early polyarthritis and homozygous for MBL structural variant alleles have a higher risk of developing early erosive RA. These findings, together with the positive association between MBL variant alleles and the increased serum levels of IgM RF and CRP, point at the MBL gene as a relevant locus in the pathophysiology of RA.

CXC chemokine receptor 3 (CXCR3), which is known to be expressed predominately on memory and activated T lymphocytes, is a receptor for both interferon ? (IFN-?)–inducible protein 10 (?IP-10) and monokine induced by IFN-? (Mig). We report the novel finding that CXCR3 is also expressed on CD34+ hematopoietic progenitors from human cord blood stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF) but not on freshly isolated CD34+ progenitors. Freshly isolated CD34+progenitors expressed low levels of CXCR3 messenger RNA, but this expression was highly up-regulated by GM-CSF, as indicated by a real-time quantitative reverse transcriptase–polymerase chain reaction technique. ?IP-10 and Mig induced chemotaxis of GM-CSF–stimulated CD34+ progenitors by means of CXCR3, since an anti-CXCR3 monoclonal antibody (mAb) was found to block ?IP-10–induced and Mig-induced CD34+ progenitor chemotaxis. These chemotactic attracted CD34+ progenitors are colony-forming units—granulocyte-macrophage. ?IP-10 and Mig also induced GM-CSF–stimulated CD34+ progenitor adhesion and aggregation by means of CXCR3, a finding confirmed by the observation that anti-CXCR3 mAb blocked these functions of ?IP-10 and Mig but not of chemokine stromal cell–derived factor 1a. ?IP-10–induced and Mig-induced up-regulation of integrins (CD49a and CD49b) was found to play a crucial role in adhesion of GM-CSF–stimulated CD34+progenitors. Moreover, ?IP-10 and Mig stimulated CXCR3 redistribution and cellular polarization in GM-CSF–stimulated CD34+progenitors. These results indicate that CXCR3–?IP-10 and CXCR3–Mig receptor-ligand pairs, as well as the effects of GM-CSF on them, may be especially important in the cytokine/chemokine environment for the physiologic and pathophysiologic events of differentiation of CD34+ hematopoietic progenitors into lymphoid and myeloid stem cells, subsequently immune and inflammatory cells. These processes include transmigration, relocation, differentiation, and maturation of CD34+ hematopoietic progenitors.

CXC chemokine receptor 3 (CXCR3), which is known to be expressed predominately on memory and activated T lymphocytes, is a receptor for both interferon γ (IFN-γ)–inducible protein 10 (γIP-10) and monokine induced by IFN-γ (Mig). We report the novel finding that CXCR3 is also expressed on CD34+hematopoietic progenitors from human cord blood stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF) but not on freshly isolated CD34+progenitors. Freshly isolated CD34+progenitors expressed low levels of CXCR3 messenger RNA, but this expression was highly up-regulated by GM-CSF, as indicated by a real-time quantitative reverse transcriptase–polymerase chain reaction technique. γIP-10 and Mig induced chemotaxis of GM-CSF–stimulated CD34+progenitors by means of CXCR3, since an anti-CXCR3 monoclonal antibody (mAb) was found to block γIP-10–induced and Mig-induced CD34+progenitor chemotaxis. These chemotactic attracted CD34+progenitors are colony-forming units—granulocyte-macrophage. γIP-10 and Mig also induced GM-CSF–stimulated CD34+progenitor adhesion and aggregation by means of CXCR3, a finding confirmed by the observation that anti-CXCR3 mAb blocked these functions of γIP-10 and Mig but not of chemokine stromal cell–derived factor 1α. γIP-10–induced and Mig-induced up-regulation of integrins (CD49a and CD49b) was found to play a crucial role in adhesion of GM-CSF–stimulated CD34+progenitors. Moreover, γIP-10 and Mig stimulated CXCR3 redistribution and cellular polarization in GM-CSF–stimulated CD34+progenitors. These results indicate that CXCR3–γIP-10 and CXCR3–Mig receptor-ligand pairs, as well as the effects of GM-CSF on them, may be especially important in the cytokine/chemokine environment for the physiologic and pathophysiologic events of differentiation of CD34+hematopoietic progenitors into lymphoid and myeloid stem cells, subsequently immune and inflammatory cells. These processes include transmigration, relocation, differentiation, and maturation of CD34+hematopoietic progenitors.

To investigate the possible association of mannose-binding lectin (MBL) genotypes with the outcome of rheumatoid arthritis (RA). MBL genotypes and plasma concentrations were retrospectively determined in 140 RA patients who were selected from a major cohort followed up prospectively for up to 32 years. MBL-insufficient patients (those with 2 defective structural MBL alleles or with 1 defective allele combined with a low-expression variant of the normal allele) had unfavorable outcomes. The relative risk of a severe radiographic outcome event (30% of maximum radiographic destruction, or an RE30) was 3.1 (95% confidence interval 1.8–5.1) in the MBL-insufficient group versus the MBL-competent group (P &lt; 0.0001). An RE30 occurred in 50% of MBL-competent patients within 17 years, while such an event occurred 9 years earlier in MBL-insufficient patients (i.e., within 8 years) (P &lt; 0.0001). During the first 15 years, there was a significant trend toward lower hemoglobin levels (P &lt; 0.04), higher erythrocyte sedimentation rates (P &lt; 0.02), and a higher number of swollen joints (P &lt; 0.05) in the MBL-insufficient group. MBL genotypes giving rise to MBL insufficiency are highly significant risk factors for fast progression of radiographic joint destruction.

To determine whether variant alleles in the coding portion of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to systemic lupus erythematosus (SLE) and concomitant infections. MBL alleles and serum concentrations were determined by polymerase chain reaction and enzyme-linked immunosorbent assay, respectively, in 91 Danish patients with SLE and in 250 controls. Homozygosity for MBL variant alleles was observed in 7.7% of the SLE patients compared with 2.8% of the controls (P = 0.06), while no difference was seen for heterozygosity (33.0% versus 34.4%). Homozygotes had an increased risk of acquiring serious infections compared with patients who were heterozygous or homozygous for the normal allele (odds ratio 8.6, 95% confidence interval 1.5–47.6, P = 0.01). The time interval from the diagnosis of SLE to the first infectious event was shorter (P = 0.017), and the annual number of infectious events was 4 times higher, among homozygotes (P = 0.00002). They were especially prone to acquire pneumonia (P = 0.00004). Homozygosity for MBL variant alleles may explain much of the increased risk of complicating infections seen in SLE patients. Additionally, it is a minor risk factor for acquiring SLE.

In the genetically homogeneous Scandinavian population, we have investigated chromosome 5 and the HLA(human leukocyte antigen) region on chromosome 6p21 by applying linkage and association analyses on 106 white sibling pair families with multiple sclerosis. The importance of genes within the HLA region for the susceptibility of multiple sclerosis has previously been reported. More recently, findings have suggested importance of regions on chromosome 5. Half of chromosome 5 was analyzed by using 14 microsatellite markers and a susceptibility region with a maximum LOD score of 1.1 was identified. Chromosome 6 was analyzed by HLA‐DRtyping and using the TNFamicrosatellite marker. A peak maximum LOD score of 2.0 was found at the HLA‐DRmarker. Association studies were made for all the markers, comparing 106 probands from the sibling pairs with 100 unrelated controls. None of the markers on chromosome 5 showed significant association with multiple sclerosis, whereas strong association between multiple sclerosis and DR2was found, with an odds ratio of 3.7 (p< 10−5). It is surprising that association was not seen for any of the TNFaalleles including the 121‐bp allele, although this allele was in positive linkage disequilibrium with DR2in both patients and controls. Our results support the existence of multiple sclerosis susceptibility loci on chromosomes 5p and 6p21.

In the genetically homogeneous Scandinavian population, we have investigated chromosome 5 and the HLA (human leukocyte antigen) region on chromosome 6p21 by applying linkage and association analyses on 106 white sibling pair families with multiple sclerosis. The importance of genes within the HLA region for the susceptibility of multiple sclerosis has previously been reported. More recently, findings have suggested importance of regions on chromosome 5. Half of chromosome 5 was analyzed by using 14 microsatellite markers and a susceptibility region with a maximum LOD score of 1.1 was identified. Chromosome 6 was analyzed by HLA-DR typing and using the TNFa microsatellite marker. A peak maximum LOD score of 2.0 was found at the HLA-DR marker. Association studies were made for all the markers, comparing 106 probands from the sibling pairs with 100 unrelated controls. None of the markers on chromosome 5 showed significant association with multiple sclerosis, whereas strong association between multiple sclerosis and DR2 was found, with an odds ratio of 3.7 (p &lt; 10⁻⁵). It is surprising that association was not seen for any of the TNFa alleles including the 121-bp allele, although this allele was in positive linkage disequilibrium with DR2 in both patients and controls. Our results support the existence of multiple sclerosis susceptibility loci on chromosomes 5p and 6p21.

This study investigated the involvement of cyclic adenosine monophosphate (cAMP) in contrast medium-induced renal vasomotor effects and the efficacy of selective phosphodiesterase (PDE) inhibitors influencing cAMP in preventing contrast medium-induced renal vasospasm.

Changes in contractility of corpus cavernosum (CC) smooth muscle caused by radio contrast medium may result in misinterpretations of cavernosography used diagnostically in erectile dysfunction.

Contrast media (CM)-induced renal vasoconstriction is an important factor in the pathogenesis of CM-induced nephrotoxicity. The effects of ionic, high-osmolar CM sodium/meglumine diatrizoate and nonionic, low-osmolar CM iohexol and iopamidol were studied in rabbit, dog, and pig renal arteries and compared with human tissue in an organ bath.

OBJECTIVE: To study the association of brain magnetic resonance imaging (MRI) findings and HLA findings to clarify the relationship between monosymptomatic optic neuritis (ON) and ON as part of clinically definite multiple sclerosis (CDMS). DESIGN: Population-based cohort of patients with ON referred prospectively during 6 years by neurologists and ophthalmologists within 4 weeks of onset of ON. SETTING: Referral center in the general community of greater Copenhagen (Denmark) (population, 1.5 million). PATIENTS: A consecutive sample of 199 patients aged 12 to 59 years with ON (133 with idiopathic ON, 66 with ON+CDMS), ethnically matched with 192 healthy volunteers. MAIN OUTCOME MEASURES: Relation between the HLA-DR15, -DR17, -DQA-1B, and -DQB-1B polymorphisms as defined by restriction fragment length polymorphism analysis, and presence of plaques on T2-weighted brain MRI. RESULTS: The frequency of HLA-DR15 was significantly increased in patients with ON+CDMS (52%) and ON (47%) compared with control subjects (31%). The frequency of HLA-DR17 was almost equal in the ON + CDMS (18%), ON (23%), and control (23%) groups. The frequencies of HLA-DQA-1B (55% in ON+CDMS, 58% in ON) and HLA-DQB-1B (49% in ON+CDMS, 59% in ON) were significantly increased compared with control subjects (41%, HLA-DQA-1B; 37%, HLA-DQB-1B). Brain MRI was abnormal in 48 of 56 examined patients with ON+CDMS and in 64 of 120 examined patients with ON (P&lt;.001). In contrast, the frequencies of HLA alleles did not differ between patients with and without demyelinating lesions. However, patients with ON and normal MRI findings did not show association with HLA-DR15. CONCLUSIONS: The frequencies of alleles were similar in patients with ON and ON+CDMS, confirming that they are not 2 immunogenetically distinct disease entities. The heterogeneity within the group of patients with ON suggests that the HLA-DR15 molecule is involved in susceptibility to initial demyelinating lesion formation.

The new quinolones have broad antimicrobial spectra covering all aerobic gram-negative and gram-positive bacteria encountered in urinary tract infections. All are administered orally, some also parenterally, low degree of resistance, few side effects and bacteriological and clinical cure rates similar to or higher than traditional antimicrobials make them especially suitable for treatment of complicated urinary tract infections including bacterial prostatitis. Noncritical use of quinolones in simple infections where standard drugs may be equally effective and safe should be discouraged.

The principles and techniques of uroflowmetry, cystometry, pressure-flow studies and urethral pressure profilometry are reviewed. The interpretation of urodynamic tests depends on the methodology used and whether the patients usual voiding symptoms during the examination. The more complicated urodynamic tests may require computer assistance both for data storage but also for test interpretation. One of the main challenge in future urodynamic is to transform these tests to clinical usable tools.

The compensatory renal hypertrophy and hyperfunction following unilateral nephrectomy have been studied in dogs over a two-year period. It was found that a sharp increase in function and volume of kidney tissue takes place during the first two weeks. CCr and volume of kidney tissue continue to increase reaching average values after two years of 88% and 92% respectively of those for two kidneys. CPAH and TmPAH remain almost stable at 65% and 57% respectively after an initial increase during the first two weeks. All values are expressed in percent of the function of two normal kidneys before unilateral nephrectomy. The filtration seems to be increasingly efficient and the functional integrity of the nephrons is preserved, although a glomerular predominance is encountered two years following unilateral nephrectomy.

Bacterial prostatitis in dogs was induced by injection of an E. coli 06 suspension into a branch of the prostatic artery.

Compensatory renal hypertrophy and hyperfunction during contralateral chronic hydronephrosis was investigated. It was found that graded impairment of the function in hydronephrotic kidneys causes proportional compensatory hypertrophy and hyperfunction in the contralateral kidney. The functional pattern both “per kidney” and “per nephron” was the same in the kidneys investigated here and in kidneys where contralateral nephrectomy had been performed, but whereas a decreased ability to concentrate the urine maximally and to reabsorb sodium was encountered in the latter, this could not be detected in this study. On the other hand, a decreased COSM in the diseased kidney did not stimulate a corresponding increase in the contralateral kidney as did unilateral nephrectomy. Adaptive and growth dependent changes in compensatory renal hypertrophy are discussed.

Monolayer tissue culture cells from benign nodular hyperplasia of the prostate transferred at 1–2 weeks intervals were examined under the electron microscope after the 3rd, 9th, and 10th transfers. Changes seen after 9 to 10 transfers were interpreted as an “aging process” and consisted of the presence of lysosomes of various types and variations in the mitochondria profile. These changes were described in detail and illustrated and compared to the ultrastructural appearance of monolayer cell cultures in the early transfer stage.

Renal function, rate of urine turnover in the renal pelvis, and the role of the renal lymphatics were studied in dogs during total ureteral occlusion lasting from 6 to 34 days with and without induced infection. — Glomerular filtration rate, effective renal plasma flow, concentration ability, and tubular reabsorption of sodium and potassium were followed. The concentrations of 2 radiolabelled clearance substances, iothalamate and o-iodohippurate, as well as those of sodium, potassium, and total protein were determined in hilar and/or subcapsular lymph obtained by cannulization of the lymphatics. — In uninfected hydronephrosis, concentration ability as well as sodium and potassium reabsorption were impaired as compared with control kidneys. In infected hydronephrosis, these partial functions were not maintained. Turnover in the occluded renal pelvis, ranged from 0.04 to 0.16 ml per minute. Following ureteral occlusion of 1 week, pyelolymphatic reabsorption into hilar, but not subcapsular lymph could be demonstrated.

Summary We investigated the effects of bladder irrigation with various irrigation fluids at various temperatures and irrigation pressures on the adherence of3H-labeledEscherichia coli O6 to bladder urothelium in a rat model. Sixty-four control and 108 experimental animals were studied. The latter were divided into five groups according to type of irrigation fluid (glycine 1.5%, sorbitol 2.7%, mannitol 0.54%, and H2O), fluid temperature, and intravesical pressure during irrigation. Only rats undergoing high pressure (60 cm H2O) bladder irrigation demonstrated significant increases in bacterial adherence compared to controls. No differences were found with the different irrigation fluids. This study suggests that high intravesical pressure during transurethral surgery might enhance bacterial adherence to urothelium and thereby increase the risk of manifest infection in the postoperative course.

Summary Constant infusion experiments were performed on dogs to determine the concentration levels of co-trimazine (a combination of trimethoprim and sulphadiazine) in the dogs' prostatic secretion, prostatic interstitial fluid, prostatic tissue, urine, and other tissues. Trimethoprim concentrations in prostatic secretion, interstitial fluid and tissue were higher than corresponding serum values, whereas the concentrations of sulphadiazine were far below these values. These results were compared to those of a similar study which combined trimethoprim and sulphamethoxazole (co-trimoxazole). The prostatic secretion to serum ratios of sulphadiazine were lower than those for sulphamethoxazole. Urine concentrations of sulphadiazine, however, were much higher than those of sulphamethoxazole. In serum and urine, only co-trimazine had a trimethoprim/sulphonamide ratio approximating the optimal synergy ratio of 1:20. Neither trimethoprim/sulphonamide combination appears to have any theoretical advantage over the other in the treatment of prostatitis. However, co-trimazine may be more effective in the treatment of urinary tract infections in general because of the high concentrations of sulphadiazine in urine.

Summary Renal clearance and renal hilar lymph and interstitial fluid concentrations of co-trimazine were investigated in dogs during constant intravenous infusion. The lymph was obtained by direct cannulation of the lymphatics, and the interstitial fluid from small plastic tissue chambers, implanted 4–6 weeks before the experiments. The clearance values for trimethoprim and sulphadiazine, compared to the glomerular filtration rate, as measured by125I-iothalamate clearance, showed a net tubular secretion of trimethoprim and tubular re-absorption of sulphadiazine. Renal lymph and interstitial fluid concentrations were found to be lower than the plasma concentrations, corresponding well with earlier findings for several other antimicrobial agents. We found a statistically significant difference between lymph/plasma and interstitial fluid/plasma ratios, indicating different composition and origin of lymph and interstitial fluid.

Summary Lipid solubility and a basic pKa are the necessary requirements for the secretion of antibacterial agents into the prostatic fluid in dogs in concentrations exceeding the simultaneous serum concentrations. Protein binding appears to be of less importance. This was demonstrated in constant infusion experiments in dogs. Ampicillin concentrations in prostatic fluid in a patient with urinary diversion were found to be much higher following oral administration of the lipid-soluble hetacillin ester than following administration of ampicillin. This had also previously been found to be true in prostatic fluid and tissue of dogs. The prostatic tissue ampicillin concentrations in humans were not found to be increased following hetacillin ester administration, possibly because the drug was washed out of the tissue during surgery. The importance of the basic pKa is shown in constant infusion experiments in dogs with trimethoprim, erythromycin and rosamicin, all 3 lipid-soluble antibiotics having a high pKa and all being found to concentrate in prostatic fluid and tissue of dogs.

Summary The renal clearance, renal lymph concentrations and serum protein binding were determined in the dog for 5 cephalosporine derivatives (cephaloridine, cephalothin, cephazoline, cephacetrile, and cephradine). The protein binding for these cephalosporines was 11%, 50%, 24%, 24%, and 10% respectively. These cephalosporines were either excreted exclusively by glomerular filtration (cephaloridine and cephradine) or in addition by tubular secretion (cephalothin, cephazoline, and cephacetrile). The renal lymph concentrations of all antibiotics were significantly lower than the simultaneous arterial plasma concentrations (mean values between 64% and 84%). There was a significant negative correlation between the renal clearance and the lymph/plasma ratio of the antibiotic. There was also an indirect negative correlation between serum protein binding and the relative renal lymph concentrations. The renal clearance, therefore, but not the protein binding, has a direct effect on the lymph concentration. The renal interstitium is divided into a perivascular and a peritubular interstitium. Since lymph vessels are only present in the perivascular connective tissue, lymph concentrations can only reflect the conditions in the perivascular interstitium. Since the post-glomerular capillaries contain pores offering permeation possibilities for even high molecular substances, good tissue permeability for free as well as protein-bound antibiotics can be assumed. The interstitial concentration depends on the capillary plasma concentration and this in turn on the renal excretion mechanism of the antibiotic.