A double blind, placebo-controlled, crossover therapy study with natural human IL-2 (nhuIL-2) in combination with regular intravenous gammaglobulin (IVIG) infusions in 10 patients with common variable immunodeficiency (CVID)

Ten CVID patients with defective IL-2 synthesis in vitro were treated with nhuIL-2 in a placebo-controlled, double blind, crossover therapy study during a period of 12 months. No severe side-effects of nhuIL-2 were recorded. Marginal serum nhuIL-2 levels were measurable in individual patients only during the therapy phase. Serum levels of soluble IL-2 receptors were unaffected by the therapy. nhuIL-2 and placebo groups did not differ significantly with respect to requirement of IVIG substitutions which were performed whenever serum IgG levels dropped below 5 g/l: a total of 53 IVIG infusions (corresponding to 17.6 g IgG/month per patient) was necessary during the placebo phase, and 48 infusions (16-4 g IgG/month per patient) during the nhuIL-2 treatment phase. Thus, nhuIL-2 therapy was ineffective in improving spontaneous IgG synthesis in vivo. Nevertheless, the group of patients receiving nhuIL-2 during the first 6 months of the study exhibited a significant reduction of severe infections (n = 25) during the following 6 months of placebo treatment (n = 7) (P < 0–045). The infection score dropped in this group from 181 to 23 (P < 0015). Patients of the second group receiving first placebo and then nhuIL-2 did not experience a significant difference in number and score of infectious episodes: 25 infections were recorded during the first 6 months and 24 during the following 6 months. We suppose that nhuIL-2 therapy of CVID patients reduces susceptibility to severe infections, possibly via the induction of a specific antibody response, which is effective at the earliest 6 months after initiating nhuIL-2 therapy.     

COMMENTS

Human spermatogenesis is regulated by a network of genes located on autosomes and on sex chromosomes, but especially on the Y chromosome. Most results concerning the germ cell function of the Y genes were obtained by genomic breakpoint mapping studies of the Y chromosome of infertile patients. Although this approach has the benefit of focussing on those Y regions that contain most likely the Y genes of functional importance, its major drawback is the fact that fertile control samples were often missing. In fertile men, molecular and cytogenetic analyses of the Y chromosome has revealed highly polymorphic chromatin domains especially in the distal euchromatic part (Yq11.23) and in the heterochromatic part (Yq12) of the long arm. In sterile patients cytogenetic analyses mapped microscopically visible Y deletions and rearrangements in the same polymorphic Y regions. The presence of a Y chromosomal spermatogenesis locus was postulated to be located in Yq11.23 and designated as AZoospermia Factor (ZF). More recently, molecular deletion mapping in Yq11 has revealed a series of microdeletions that could be mapped to one of three different AZF loci: AZFa in proximal Yq11 (Yq11.21), AZFb and AZFc in two non‐overlapping Y‐regions in distal Yq11 (Yq11.23). This view was supported by the observation that AZFa and AZFb microdeletions were associated with a specific pathology in the patients' testis tissue. Only AZFc deletions were associated with a variable testicular pathology and in rare cases AZFc deletions were even found inherited from father to son. However, AZFc deletions were found with a frequency of 10–20% only in infertile men and most of them were proved to be “de novo”, i.e. the AZFc deletion was restricted to the patient's Y chromosome. Based mainly on positional cloning experiments of testis cDNA clones and on the Y chromosomal sequence now published in GenBank, a first blueprint for the putative gene content of the AZFc locus can now be given and the gene location compared to the polymorphic DNA domains. This artwork of repetitive sequence blocks called AZFc amplicons raised the question whether the AZFc chromatin is still part of the heterochromatic domain of the Y long arm well known for its polymorphic extensions or is decondensed and part of the Yq11.23 euchromatin? We discuss also the polymorphic DAZ gene family and disclose putative origins of its molecular heterogeneity in fertile and infertile men recently identified by the analyses of Single Nucleotide Variants (SNVs) in this AZFc gene locus.     

Programmed Cell Death of Peripheral Blood B Cells Determined by Laser Scanning Cytometry in Sjögren’s Syndrome with a Special Emphasis on BAFF

Functionally impaired B cells play an important role in the pathogenesis of Sjögrens syndrome (SS). The aim of the study was to investigate the apoptosis susceptibility of peripheral blood B cells from patients with SS and the impact of B cell activating factor (BAFF) on the apoptosis capability of these cells in correlation with IgG production. Peripheral blood B cells were isolated and stained for apoptosis markers (Bax, Bcl-2) and members of the TNF-R superfamily, CD95 and CD40. The apoptosis frequency of cells bearing these markers were assessed. Also, the apoptosis capability of cultured B-lymphocytes was investigated in medium alone, with anti-CD95 or with soluble BAFF. Quantitative ELISA was performed to detect plasma levels of sBAFF. Furthermore, the level of circulating B-cell cytokines was measured. BAFF levels were compared between patients with normal and elevated IgG levels. In SS, Bcl-2 positive B cell counts were significantly higher then in controls, also in this population the apoptosis frequency was reduced. Apoptosis within Bax+ and CD40+ B cells were significantly decreased in patients. BAFF induced a significant antiapoptotic effect in SS; also this effect was clearly evident in B cells from SS with hypergammaglobulinaemia. Plasma BAFF levels were significantly higher in SS, mostly in patients with hypergammaglobulinaemia. Plasma B-cell cytokines were raised in SS. In Sjögrens syndrome B cells, a general antiapoptotic tendency might lead to prolonged B-cell survival driven at least partly by elevated levels of BAFF and supposedly by B-cell cytokines. Also, the exaggerated BAFF stimulation might lead to excessive immunoglobulin production. The B-cell apoptosis defects, the increased BAFF levels—correlating with hypergammaglobulinaemia—together with the raised B-cell cytokine levels indicates the disturbed B-cell biology in the disease.     

Regional differences in cortical electroencephalogram (EEG) slow wave activity and interhemispheric EEG asymmetry in the fur seal

Slow wave sleep (SWS) in the northern fur seal (Callorhinus ursinus) is characterized by a highly expressed interhemispheric electroencephalogram (EEG) asymmetry, called ‘unihemispheric’ or ‘asymmetrical’ SWS. The aim of this study was to examine the regional differences in slow wave activity (SWA; power in the range of 1.2–4.0 Hz) within one hemisphere and differences in the degree of interhemispheric EEG asymmetry within this species. Three seals were implanted with 10 EEG electrodes, positioned bilaterally (five in each hemisphere) over the frontal, occipital and parietal cortex. The expression of interhemispheric SWA asymmetry between symmetrical monopolar recordings was estimated based on the asymmetry index [AI = (L?R)/(L+R), where L and R are the power in the left and right hemispheres, respectively]. Our findings indicate an anterior–posterior gradient in SWA during asymmetrical SWS in fur seals, which is opposite to that described for other mammals, including humans, with a larger SWA recorded in the parietal and occipital cortex. Interhemispheric EEG asymmetry in fur seals was recorded across the entire dorsal cerebral cortex, including sensory (visual and somatosensory), motor and associative (parietal or suprasylvian) cortical areas. The expression of asymmetry was greatest in occipital–lateral and parietal derivations and smallest in frontal–medial derivations. Regardless of regional differences in SWA, the majority (90%) of SWS episodes with interhemispheric EEG asymmetry meet the criteria for ‘unihemispheric SWS’ (one hemisphere is asleep while the other is awake). The remaining episodes can be described as episodes of bilateral SWS with a local activation in one cerebral hemisphere.     

c-myc GENE ABNORMALITIES IN MUCOSA-ASSOCIATED LYMPHOID TISSUE (MALT) LYMPHOMAS

c-myc gene abnormalities associated with lymphomagenesis, including rearrangements and mutations in the regulatory region between exon I and intron I, have been studied in 54 MALT lymphomas (43 low and 11 high grade) and 36 nodal lymphomas (27 low and 9 high grade). By Southern blot analysis, none of the 54 MALT lymphomas but 2 of 36 nodal lymphomas had c-myc gene rearrangements. Defined tumour cell populations from all MALT lymphoma cases were isolated by microdissection from frozen tissue sections and analysed by polymerase chain reaction–single-strand conformational polymorphism (PCR–SSCP) and direct sequencing for somatic mutations in the exon I/intron I region of the gene. Point mutations in this region were identified in nine cases of MALT lymphoma (7/43=16·2 per cent of low grade; 2/11=18·1 per cent of high grade). These mutations were located at either the exon I/intron I border of myc intron factor (MIF) binding sites, which are critical in the negative regulation of c-myc expression. Of the nodal lymphomas, only the two cases (5·6 per cent) with c-myc gene rearrangement showed scattered or clustered mutations. These results suggest that c-myc mutations in MALT lymphomas are unlikely to be associated with chromosome translocation, which is the main cause of somatic mutations observed in other types of lymphoma. The mutations involving the c-myc regulatory regions may play a pathogenetic role in at least a proportion of MALT lymphomas. © 1997 by John Wiley & Sons, Ltd.     

Anesthetic management of the hybrid stage 1 procedure for hypoplastic left heart syndrome (HLHS)

Introduction: Despite advances in the surgical and perioperative management of patients with hypoplastic left heart syndrome (HLHS), outcomes for this high‐risk group of patients remains suboptimal. The hybrid approach [bilateral pulmonary artery (PA) banding, ductal stenting, balloon atrial septostomy], is an emerging alternative therapy for the management of HLHS, which defers the risks of a major surgical repair until the infants are older. This article will describe our experience providing the anesthetic management of patients undergoing the hybrid procedure. Methods: After Institutional Review Board approval, we retrospectively reviewed the records of 77 patients who underwent the hybrid procedure as neonates between July 2002 and August 2008. We reviewed both the anesthetic and intensive care records. Results: The hybrid procedure was performed in 77 patients (31 female and 46 male). The average age of the patients was 11.8 days with an average weight of 2.98 kg. Fentanyl was used for analgesia at an average dose of 5.7 mcg·kg^?1. The average increase in the systolic blood pressure after placement of the right and left PA bands was 11.3 mmHg. The average drop in the systemic saturation after placement of the bands was 7%, with an average postband and stent SaO₂ of 82%. Twenty‐one patients received blood transfusion (27.3%) at an average dose of 43.5 ml (14.5 ml·kg^?1). Forty patients received albumin during the case (51.9%) at an average dose of 23.2 ml (7.7 ml·kg^?1). Seventeen patients arrived at the hybrid suite already intubated, and no attempt was made to extubate these patients at the end of the case. Thirty‐six patients were extubated at the end of the procedure, and a total of 64.9% of patients were extubated within the first 24 h postoperatively. Patients had notably stable hemodynamics throughout the first 24 h in the intensive care unit. Discussion: Patients undergoing the hybrid procedure have relatively stable intraoperative and early postoperative hemodynamics. The procedure is performed without cardiopulmonary bypass (CPB) and with minimal narcotic and anesthetic exposure. Patients typically do not require blood transfusions or inotropic support and are extubated at either the end of the procedure or within 24 h of ICU admission. In our experience, the anesthetic management of patients undergoing the hybrid procedure is straightforward and requires relatively few interventions when compared to traditional neonatal surgical repairs. Deferring the risks of anesthesia, CPB, hypothermic circulatory arrest, and prolonged postoperative sedation may yield developmental advantages to patients born with HLHS.     

A functional study of low molecular weight IgM from patients with autoimmune disease

High levels of low molecular weight (LMW) IgM in certain diseases are associated with clinical and laboratory indices which reflect the severity of the disease. These associations suggest that LMW IgM may play an important role in the immunopathogenesis of these diseases. To further approach the question concerning the functional activity of LMW IgM in disease, a panel of LMW IgM and high molecular weight (HMW) IgM preparations with or without rheumatoid factor (RF) activity were used to investigate their antibody binding activity and their effector function. It was found that LMW IgM-RF and HMW IgM-RF had a similar binding capacity to Fc fragment as there was no significant difference in the affinity index between them. It further showed that the rate of activation and total amount of utilization of complement by LMW IgM and HMW IgM was similar, although the mean fluorescence of C3 deposition by IgM-RF and HMW IgM-RF was slightly higher than that of LMW IgM-RF and other control RF antibodies. However, the current study demonstrated that LMW IgM had strong neutrophil activating properties when compared with HMW IgM. These findings suggest that one mechanism of LMW IgM contributing to the immunopathogenesis of RA may be due to the formation of circulating immune complex ( CIC) by LMW IgM with subsequent activation of neutrophils. Whether LMW IgM has other functional activity in disease is unclear and needs further investigation.     

Synthesis of N-oxide derivatives of metyrapone and their detection as in vitro metabolites†

A variety of possible N-oxidation products of 2-methyl-1, 2-bis(3-pyridyl)propan-1-one (metyrapone) have been synthesized by peracid oxidation, and characterized using various spectroscopic techniques. Specific and sensitive chromatographic techniques have been developed for the separation and identification of its in vitro metabolites. Incubation of metyrapone with rat or mouse hepatic microsomes, in the presence of a NADPH-regenerating system, leads to the formation of metyrapol (keto-reduction), and two mono-N-oxides.     

Patterns of Granulocyte Kinetics in Health, Infection and in Carcinoma

Leukokinetic studies were performed using granulocytes labeled in vitrowith radioactive diisopropylfluorophosphate (DFP³²). The half-time of thegranulocytes in the circulation, blood granulocyte mass and granulocyteturnover rates were determined.

In control subjects the mean half-life was 6.44 hours with a range of 5.1 to7.7 hours. The mean blood granulocyte mass was 38 x 10⁹ cells with a rangeof 19.9 to 36.4 x 10⁹ cells and the granulocyte turnover rate was 4.08 x 10⁹ cellsper hour with a range of 2.51 to 5.50 x 10⁹ cells per hour. There was a directrelationship between the half-life and the blood granulocyte mass in thecontrol subjects.

In 6 subjects with infection the blood granulocyte mass was uniformlyincreased. The mean half-life and mean granulocyte turnover rate were bothincreased above the normal range.

In 11 subjects with carcinoma several different leukokinetic patterns werefound. The blood granulocyte mass was raised in 5 patients, but in only one ofthese was the granulocyte turnover rate increased above the normal range.In 6 subjects the blood granulocyte mass was within the normal range anddeviations from the mean control value were accompanied by proportionatechanges in the granulocyte turnover rate in all but 1 patient.

No relation was found between the half-life and the blood granulocyte massin subjects with infection and/or carcinoma. The possibility that this was dueto the establishment of a new steady state of blood granulocyte mass at alteredlevels of granulocyte production, or that steady state conditions did not existhas been considered. However the data are interpreted no evidence forsuppressed granulopoiesis was found in subjects with advanced malignantdisease.

Submitted on May 8, 1964 Accepted on July 26, 1964     

HORMONAL CHANGES ASSOCIATED WITH TESTICULAR ATROPHY AND GYNAECOMASTIA IN PATIENTS WITH LEPROSY

Basal LH, FSH, 17 beta-oestradiol and testosterone and the gonadotrophin responses to luteinizing hormone releasing hormone (LHRH) were studied in male patients with leprosy (twenty-four with lepromatous and six with tuberculoid leprosy). The mean basal LH and FSH was significantly elevated in the lepromatous group and was associated with an excessive response of both gonadotrophins following LHRH administration. The mean basal testosterone and 17 beta-oestradiol values in the lepromatous group were significantly lower than those of the tuberculoid and control groups. The abnormal gonadotrophin and sex steroid values in the lepromatous group are in keeping with the testicular atrophy and gynaecomastia accompanying this form of leprosy. However, the lack of a significant correlation between basal FSH and testicular atrophy should be noted. In addition, no correlation between any of these hormonal values and gynaecomastia could be demonstrated. The patients with tuberculoid leprosy had essentially normal hormonal profiles (except for two who had raised 17 beta-oestradiol values). This is compatible with the lack of gonadal involvement in these patients.