Safety, tolerability, and pharmacodynamics of an anti-interleukin-1α/ß dual variable domain immunoglobulin in patients with osteoarthritis of the knee: a randomized phase 1 study.

OBJECTIVE: To investigate the safety, tolerability, pharmacokinetics, and pharmacodynamics of ABT-981, a human dual variable domain immunoglobulin simultaneously targeting interleukin (IL)-1α and IL-1ß, in patients with knee osteoarthritis (OA). METHOD: This was a randomized, double-blind, placebo-controlled, single-center study of multiple subcutaneous (SC) injections of ABT-981 in patients with mild-to-moderate OA of the knee (NCT01668511). Three cohorts received ABT-981 (0.3, 1, or 3 mg/kg) or placebo every other week for a total of four SC injections, and one cohort received ABT-981 (3 mg/kg) or placebo every 4 weeks for a total of three SC injections. Assessment of safety and tolerability were the primary objectives. A panel of serum and urine biomarkers of inflammation and joint degradation were evaluated. RESULTS: A total of 36 patients were randomized (ABT-981, n = 28; placebo, n = 8); 31 (86%) completed the study. Adverse event (AE) rates were comparable between ABT-981 and placebo (54% vs 63%). The most common AE reported with ABT-981 vs placebo was injection site erythema (14% vs 0%). ABT-981 significantly reduced absolute neutrophil count and serum concentrations of IL-1α/IL-1ß, high-sensitivity C-reactive protein, and matrix metalloproteinase (MMP)-derived type 1 collagen. Serum concentrations of MMP-derived type 3 collagen and MMP-degraded C-reactive protein demonstrated decreasing trends with ABT-981. Antidrug antibodies were found in 37% of patients but were not associated with the incidence or severity of AEs. CONCLUSION: ABT-981 was generally well tolerated in patients with knee OA and engaged relevant tissue targets, eliciting an anti-inflammatory response. Consequently, ABT-981 may provide clinical benefit to patients with inflammation-driven OA.

[Surveillance for bacteremia related to vascular access in hemodialysis]. / Sorveglianza delle infezioni batteriche ematiche e correlate all'accesso vascolare in dialisi.

Infections occur frequently in chronic dialysis patients. An incidence study of 13 months' duration (December 2003-January 2004) on nosocomial bacterial infections was performed in a dialysis center (Rivoli Hospital) in the Piedmont region of northern Italy. Data were collected according to a modified CDC methodology (CDC Dialysis Surveillance Network). The total incidence of infections was 4.55/100 patients/month and was lower than that reported for other reference series (CDC 6.16/100 patients/month). The low incidence was associated with a greater use of low-risk vascular access (fistula) than in other countries such as the U.S. The infection rates were: vascular access 1.73% (1.55% local infections, 0.18% access-related bacteremia); 0.82% wound infections; 0.91% pneumonia; 1% urinary tract infections; 0.09% other bacteremias. The hospital utilization rate was 6.9/100 patients/month. Blood cultures were performed in only 28.4% of patients who had received a course of antibiotics. Antibiotics were widely used but the use of vancomycin was less than reported elsewhere (e.g., CDC surveillance). The prevalence of methicillin-resistant Staphylococcus aureus (28 strains isolated) was high (>50%), whereas no vancomycin-resistant enterococci were isolated. Preventing nosocomial bacterial infections in chronic dialysis patients is a top priority; therefore, a surveillance system along the lines of the CDC model should be adopted by all dialysis centers. The risk of nosocomial infection is strongly associated with the type of vascular access.

The role of calcium in the regulation of [3H]hemicholinium-3 binding sites in rat brain.

The role of calcium in the regulation of sodium-dependent high-affinity uptake of choline was assessed in vitro in slices of the rat brain, by measuring the specific binding of [3H]hemicholinium-3 ([3H]HCh-3) and the uptake of [3H]choline. Depolarization with potassium of slices of hippocampus, cortex, or striatum significantly increased the specific binding of [3H]HCh-3 when compared to control slices. However, the observed potentiation of specific binding of [3H]HCh-3 was markedly inhibited by the removal of calcium from the incubation medium in cortex or hippocampus, but not in slices of striatum. Alterations in the uptake of [3H]choline directly paralleled the observed changes in the specific binding of [3H]HCh-3 in striatum of the rat and were unaffected by the reduction of calcium in the incubation medium. The inorganic calcium channel antagonists, cadmium and cobalt, but not magnesium, zinc, manganese or lanthanum, significantly inhibited the 40 mM potassium chloride-induced stimulation of the binding of [3H]HCh-3 in the striatum. Finally, the calcium ionophore A23187 significantly increased the binding of [3H]HCh-3 in slices of striatum, either in the presence or absence of calcium in the bathing medium. This study demonstrates regional differences in the role of extracellular calcium in the regulation of the uptake of choline and suggests the involvement of intracellular release of calcium in the in vitro regulation of the sodium-dependent high-affinity uptake of choline in the striatum.

Splicing variability in HIV type 1 revealed by quantitative RNA polymerase chain reaction.

A quantitative RNA-polymerase chain reaction (PCR) method able to detect the majority of mRNAs produced by human immunodeficiency virus type 1 (HIV-1) was developed and used to study expression of different HIV-1 clones in human cells. Amplified mRNAs were compared to known cDNA standards. This comparison permitted the optimization of PCR conditions and eliminated the generation of artifactual PCR bands. The use of RNA and cDNA standards demonstrated that the RNA amplification is linear within the tested range and suggested that it can be used to quantitate individual mRNAs. The results demonstrate the overall conservation of splicing in different HIV-1 clones. Although, in general, splicing was conserved, extensive qualitative and quantitative variability was observed in different HIV-1 clones. This variability is likely one determinant of the biological characteristics of the different HIV-1 clones, and demonstrates a great plasticity of the HIV-1 genome. The described RNA-PCR methodology was used for the study of HIV-1 expression in unstimulated peripheral blood mononuclear cells (PBMCs) of infected individuals. In general, the same mRNAs were identified in HIV-infected cultured cell lines and in unstimulated PBMCs. Analysis of a variant band found after amplification of PBMC RNA from an HIV-infected individual revealed a new splice site for the generation of Rev/Nef-encoding mRNAs. The availability of a sensitive, rapid, and essentially quantitative method to examine the major HIV-1 mRNAs will facilitate the detailed analysis of HIV-1 expression in human cells.

Analysis of human immunodeficiency virus type 1 mRNA splicing patterns during disease progression in peripheral blood mononuclear cells from infected individuals.

HIV-1 produces more than 20 mRNAs encoding the viral proteins. We have used a sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) approach to determine HIV-1 transcriptional patterns during the course of viral infection in unstimulated peripheral blood mononuclear cells (PBMCs) from different patients. Several sets of PCR primers, used in parallel reactions, allowed the amplification and specific detection of almost all individual HIV-1 transcripts. We investigated the transcriptional profile in two individuals during primary acute and early chronic infection. In these individuals, HIV-1 mRNA expression was elevated at the first time points examined and declined over time. In addition, we performed a detailed study of HIV-1 expression in several individuals over a minimum of 7 years following seroconversion. We found that long-term asymptomatic individuals had undetectable or low levels of the three classes of HIV-1 transcripts (unspliced, singly spliced, and multiply spliced). Individuals who demonstrated disease progression showed either a general increase in the amount of expression of all transcripts or elevated levels of unspliced transcripts in late-stage disease. The splicing pattern in each patient was conserved over the years and differed among the different individuals. No evidence of major changes in the splicing pattern was found during disease progression within the same individual. Thus, HIV-1 transcriptional patterns are viral strain specific rather than disease stage specific. These results indicate that high-level expression of any class of HIV-1 transcripts is associated with clinical progression. Our analysis also demonstrates the importance of using more than one set of primers to evaluate HIV-1 RNA expression, since virus in patient PBMCs showed sequence heterogeneity in conserved regions.

Involvement of phospholipase A2 in the regulation of [3H]hemicholinium-3 binding.

We have examined the effects of exogenous phospholipase A2 (PLA2) on the sodium-dependent high-affinity choline uptake mechanism as assessed by the specific binding of [3H]hemicholinium-3 ([3H]HCh-3). Incubation of striatal synaptic membranes with bee venom PLA2 resulted in a concentration-dependent increase in the specific binding of [3H]HCh-3. The effect of PLA2 on [3H]HCh-3 binding was inhibited by quinacrine, a PLA2 inhibitor, and by removal of calcium. Scatchard analysis revealed that the observed changes in binding reflected a 2-fold increase in both the capacity and affinity of [3H]HCh-3 for its binding site. Choline and N-butylcholine inhibited the specific binding of [3H]HCh-3 in both control and PLA2-treated membranes with similar potency. When a low concentration of PLA2 was incubated with the striatal synaptosomes, a small but significant increase in high-affinity [3H]choline uptake was observed. However, higher concentrations of PLA2, which further increased the specific binding of [3H]HCh-3, caused a reduction of [3H]choline uptake, apparently due to disruption of synaptosomal integrity by PLA2. Finally, potassium depolarization- and PLA2-induced increases in specific [3H]HCh-3 binding were not additive. These results suggest a possible role for endogenous PLA2 in the calcium-dependent regulation of sodium-dependent high-affinity choline uptake.

Primary basaloid squamous cell carcinoma of the trachea.

Basaloid squamous cell carcinoma (BSCC) is an unusual but well-established entity, primarily in the upper aerodigestive tract, composed of basaloid cells associated with dysplastic to neoplastic squamous cells, and characterized by an aggressive and rapidly fatal course. To date, BSCC originating in the trachea has not been reported. In this article, the authors describe the clinical, gross, and microscopic pathologic, histochemical, and immunohistochemical features of two cases of BSCC arising in the trachea. Features distinguishing this entity from other, more common tumors of the trachea, including metastases and direct extension from other sites of origin, are also discussed.

Aggressive multimodality therapy for malignant pleural mesothelioma.

Nineteen patients with clinical stage I malignant pleural mesothelioma were treated with aggressive multimodality therapy. Nine patients underwent pleurectomy and decortication followed by immediate intrapleural chemotherapy with cisplatin and mitomycin C. Ten patients required pleuropneumonectomy followed within 1 week to 2 weeks by intrapleural administration of cisplatin (100 mg). Four to 8 weeks after operation, 15 patients underwent postoperative adjuvant cisplatin-based systemic chemotherapy. There were three postoperative complications (16%) requiring reoperation and one postoperative death (5%). Intrapleural chemotherapy was well tolerated with no complications. Systemic chemotherapy was poorly tolerated, and there was one chemotherapy-related death. Sixteen patients (84%) experienced good to excellent palliation. Three patients are currently alive with no evidence of recurrent disease at 10, 35, and 43 months. The median overall survival was 13 months and the median disease-free survival, 11 months. Overall and disease-free 3-year survivals were 17% and 22%, respectively. Patients with epithelial malignant pleural mesothelioma had significantly better overall survival (p = 0.037) and disease-free survival (p = 0.02) than patients with sarcomatous or biphasic malignant pleural mesothelioma. We conclude that despite major toxicity, in select patients with clinical stage I malignant pleural mesothelioma, aggressive multimodality therapy offers effective palliation and occasional long-term disease-free survival.

Effects of calmodulin antagonists on sodium-dependent high-affinity choline uptake.

The effects of calmodulin (CaM) antagonists were investigated on the sodium-dependent high-affinity choline uptake (SDHACU) as assessed by the specific binding of [3H]hemicholinium-3 ([3H]HCh-3) and high-affinity [3H]choline uptake. Potassium depolarization caused a significant 2-fold increase in the specific binding of [3H]HCh-3 in slices of rat striatum in vitro. CaM antagonists, including trifluoperazine (TFP), W-5, W-7, promethazine and haloperidol, dose-dependently inhibited potassium depolarization-stimulated [3H]HCh-3 binding with IC50s of 20, 40, 70, 30 and 48 (microM), respectively. Scatchard analysis revealed that the inhibitory effect of TFP resulted from a decrease in Bmax but no change in Kd of [3H]HCh-3 binding. Potassium depolarization of slices also stimulated high-affinity [3H]choline uptake, which was completely inhibited by 10 microM TFP. These results are discussed in relation to the regulatory mechanisms of SDHACU.

Sex steroids and the development of the newborn mouse hypothalamus and preoptic area in vitro: III. Effects of estrogen on dendritic differentiation.

Gonadal steroids have previously been shown to enhance neuritic outgrowth from mouse preoptic area explants in vitro. To examine effects within the explant proper, dendritic fields of Golgi-stained neurons of estradiol-treated and untreated cultures were compared quantitatively. Neurons of gonadal steroid-treated cultures had significantly more dendrites arising from the cell body. Since bifurcation probability did not differ between the treatments, the primary effect of estradiol treatment appears to be induction of dendritic branches from the soma. This indicates that intra-explant dendrites, like extra-explant neurites, proliferate in response to estradiol treatment and suggests the involvement of hormone-stimulated dendrogenesis in sexual differentiation of the brain.

[3H]hemicholinium-3 binding in rats with status epilepticus induced by lithium chloride and pilocarpine.

The specific binding of [3H]hemicholinium-3 ([3H]HCh-3) and high-affinity [3H]choline uptake were measured in rats with status epilepticus induced by lithium and pilocarpine. The specific binding of [3H]HCh-3 in cortex and hippocampus from rats with status epilepticus increased to 2- to 3-fold of control while the striatal [3H]HCh-3 binding increased minimally. Scatchard analyses revealed that the observed changes resulted from an increase in Bmax of [3H]HCh-3 binding. High-affinity [3H]choline uptake remained unchanged. These results further implicate phospholipase A2 in the regulation of [3H]HCh-3 binding sites.

Rapid in vitro modulation of [3H]hemicholinium-3 binding sites in rat striatal slices.

The effects of depolarizing concentrations of potassium chloride on the modulation of [3H]hemicholinium-3 binding sites and high affinity choline uptake were examined in vitro. When rat striatal slices were incubated in Krebs buffer for 20 min, [3H]hemicholinium-3 binding sites diminished to 60% of binding measured in fresh un-incubated tissue, and remained stable for 60 min. Upon addition of Krebs buffer containing 40 mM KCl, the number of binding sites increased during a 20 min period, and remained stable for 40 min. Changes in [3H]hemicholinium-3 binding sites closely paralleled changes in high affinity choline uptake. Scatchard analysis revealed that changes in binding result from alterations in the number of binding sites (Bmax), and not in the affinity (KD). These results suggest that neuronal depolarization rapidly alters the velocity of choline transport into cholinergic neurons by increasing the number of available carriers.

Rare pulmonary manifestation of Ehlers-Danlos syndrome.

Ehlers-Danlos syndrome (EDS) is an inherited disorder of connective tissue with multiple thoracic manifestations. We present an unusual thoracic manifestation of EDS consisting of parenchymal cysts and fibrous and fibroosseous nodules. These manifestations may be related to an abnormal attempt at repair of parenchymal or vascular tears.

[Anti-HCV in patients undergoing dialysis in Piedmont and in Valle d'Aosta]. / Anti-HCV nei pazienti in dialisi in Piemonte ed in Valle d'Aosta.

Special questionnaires were distributed in order to assess the current incidence of anti-HCV antibodies in Dialysis Centres in Piedmont and the Aosta Valley. The analysis of findings showed an overall positivity of 23.9%, with higher values in those cases treated with extracorporal methods and lower values in those patients treated with peritoneal dialysis. The data also confirmed the correlations between anti-HCV antibodies and the number of transfusions performed, and suggested the possible influence of previous operations, high risk manual occupation, or more than one of these factors, in addition to environmental conditions. Lastly, the paper reports the prophylactic measures currently in use for patients and medical staff in the various centres in Piedmont and the Aosta Valley.

[Clinical and epidemiologic characteristics of selected long-term survivors undergoing dialysis]. / Caratteristiche cliniche ed epidemiologiche di pazienti selezionati da una lunga sopravvivenza in dialisi.

At the end of December, 1993, 389 patients treated by means of dialysis for more than 15 years were registered in the Piedmont Dialysis and Transplantation Register; among them, 183 were alive. Characteristics of these patients as age at the beginning of dialytic treatment, casual nephropathies, causes of death, high risk conditions, dialytic schedules, vascular access, hospitalization requirements, rehabilitation, dialysis efficacy, gross mortality, were compared with those of patients treated for a shorter time, concluding that a very long dialytic survival can be achieved with a good clinical and social rehabilitation.

[Comorbidity factors in the dialysis registries and the experience of the Piedmontese Registry]. / I fattori di comorbidità nei Registri di Dialisi e nell'esperienza del Registro Piemontese.

The parameters used at present by the Dialysis and Transplant Registries of various countries to evaluate dialyzed patients' comorbidity show great differences, mostly owing to the different epidemiological, social and racial characteristics of the studied populations. Moreover, the typology of the dialyzed patient is changing: the mean age is increasing, patients with high-risk conditions as vasculopathy and diabetes are widely accepted to the treatment. Thus the Piedmont Registry will be implemented as follows: new fields about comorbidity for clinical (blindness, cachexy and dementia), social (smoking, alcohol and drugs addiction) and diagnostic (type of diabetes, of neoplasm, of cardiovascular problem) parameters, and questions needing dichotomic response (vasculopathy yes or no) will be added. The exact time of appearance of any risk factor will be requested, and a field for "others" risk factors will be added as well, trying to deeply identify the dialytic population not affected by any comorbidity factor. Finally, a new improved control system of the collected data will be used: our registry needs to be implemented in the future by such evaluations, to go on giving useful informations about epidemiology of the dialyzed patients.

[Survival of patients starting dialysis at an advanced age in the Piedmont]. / La sopravvivenza del paziente entrato in dialisi con un'età molto avanzata in Piemonte.

BACKGROUND: Patients starting dialysis in old age (age > 70 and > 80) in Piedmont are widely increasing: the survival curves of this group of patients can give very important information to evaluate the quality of both the delivered therapy and our very wide criteria of acceptance to the treatment. To this end, using data from the Piedmont Dialysis and Transplant Register, the survival curves of patients with age over 70 and 80, beginning dialysis in all Piedmont Dialysis Units between 1981 and 1996, have been examined. METHODS: These curves have been considered both in a general way and according to the presence or absence of further high risk conditions; they show results better than expected and improving from 1981 to 1995. RESULTS: If the survival curves of these patients are considered according to the kind of dialytic treatment performed, they do not show any significative difference. CONCLUSIONS: The conclusion is drawn that these data strongly support first, the fitness of criteria of very wide acceptance to the treatment and modulated choice of the kind of dialytic treatment at present followed in Piedmont; and second, that dialysis treatment can give very good results also in elderly patients. So, it is suggested that the economic and structural difficulties of dialysis Units must not influence the nephrologist's choice towards elderly patients.

[Helicobacter pylori in patients undergoing periodic hemodialysis]. / L'Helicobacter pylori in pazienti sottoposti ad emodialisi periodica.

Forty-nine haemodialyzed patients have been submitted consecutively, under informed consent, to endoscopy with multiple antral gastric mucosa biopsies for Helicobacter pylori (HP) identification, performed by urease, microscopic and cultural tests, as well as histologic examination. Patients have been considered HP negative when negative for all tests; positivity for HP has been correlated with gastritis histologically evaluated according to Whitehead; at endoscopy, blood samples for HP specific IgG, IgA, IgM have been collected; patient's life style concerning smoke, alcohol and drugs as FANS has been investigated as well. HP prevalence in our haemodialyzed patients is 38.8 per cent, similar to general population submitted to endoscopy; a statistically significant correlation between HP and gastritis and specific IgG, but no correlation between HP and age, dialysis duration, IgA, IgM, smoking, alcohol or drugs consumption has been found.

[Social rehabilitation, state of the disease and quality of life. What are the parameters to evaluate rehabilitation of the patient under dialysis in the '90s?]. / Riabilitazione sociolavorativa, stato di malattia e qualità di vita. Quali indicatori per il paziente in dialisi negli anni '90?

The parameters used at present by the Piedmont Dialysis and Transplant Register to evaluate rehabilitation and quality of life of the dialyzed patients have been useful in the past, but they will no longer be so in the future, as the typology of the dialyzed patient is changing: the mean age is increasing, patients with high-risk conditions as vasculopathy and diabetes are widely accepted to the treatment. Thus, rehabilitation has to be evaluated not only from a social and professional viewpoint, but also as "illness situation" and "quality of life" and this can be done using the Karnofsky scale and the test of Spitzer: our Register needs to be implemented in the future by such evaluations, to go on giving us useful information about rehabilitation of the dialyzed patients.

[The status of peritoneal dialysis in Piedmont by 12/31/1997]. / La situazione della dialisi peritoneale in Piemonte al 31/12/1997.

BACKGROUND: The aim of the study is to examine the situation of chronic uremia substitutive treatment by means of peritoneal dialysis in Piedmont on December 31, 1997 using data from the Piedmont regional dialysis and transplant register. METHODS: Starting from the year 1981, data are reported (absolute, per million population, and according to different patient's anagraphic ages) about peritoneal dialysis prevalence and incidence; about basic nephropathies; drop-out from treatment; patient's rehabilitation; complications incidence, particularly peritonitis; patient's survival compared with survival of patients treated, during the same years with extracorporeal techniques. RESULTS: The data demonstrate, during these years, an increase of peritoneal dialysis incidence and prevalence; patients' survival curves, compared with those of patients treated with extracorporeal techniques, are very similar during the first years of treatment and worse afterwards, but never reaching statistical significance. CONCLUSIONS: They support Peritoneal Dialysis as a very good kind of substitutive treatment for some years, but this opinion deserves further examination and investigation on a longer period of time.