The risk of interstitial lung disease during biological treatment in Japanese patients with psoriasis.

BACKGROUND: With the increasing use of biological agents for the treatment of psoriasis, the numbers of patients with interstitial lung disease (ILD) associated with biologics have also increased. Many of these cases were associated with tumour necrosis factor (TNF)-α inhibitors, but cases associated with other families of biologics have also been reported in Japan. AIM: To analyse the background factors of patients who developed ILD, and to discuss better management of biological treatment. METHOD: We reviewed 246 patients with psoriasis who were treated with biological agents in our department to identify any pulmonary adverse events (AEs). Data on patients who developed ILD were extracted to analyse background factors, clinical type of psoriasis, time to onset of ILD, pre-existing ILD, smoking habit and prescribed drugs. RESULTS: Pulmonary AEs were seen in 22 cases, of which 11 were diagnosed as drug-induced ILD. The causative drugs were mainly TNF-α inhibitors, accounting for eight cases (six treated with infliximab, two with adalimumab). The remaining three cases were associated with secukinumab, ustekinumab and ixekizumab (n = 1 each). Notably, these three cases also had a history of drug-induced ILD. CONCLUSION: Patients with a history of drug-induced ILD seem to be more susceptible to developing another ILD induced by biologics, even if treated with interleukin-17 inhibitors. Thorough screening of risk factors and evaluation for eligibility, and careful monitoring during treatment are the best solutions to avoid serious pulmonary AE. Early detection and precise diagnosis of pulmonary AEs, especially differentiation from infectious diseases, is essential for managing biological treatment.

Clinical characteristics classified by the serum KL-6 level in patients with organizing pneumonia.

BACKGROUND: The serum Krebs von der Lungen-6 (KL-6) level is a useful marker correlated with the severity of various interstitial lung diseases. There have been few reports about the clinical characteristics of organizing pneumonia (OP) associated with the serum KL-6 levels. OBJECTIVE: This study was performed to determine whether the serum KL-6 levels can help determine the optimal treatment for OP. DESIGNS: Patients diagnosed with OP by clinical, radiological and histopathological findings were retrospectively reviewed. The OP patients were classified into two groups based on their serum KL-6 levels: normal KL-6 and high KL-6 groups. The two groups were compared with regard to their clinical and radiological data and therapeutic response one month after the start of treatment. RESULTS: The clinical records of twenty-two patients diagnosed with OP were reviewed. The serum KL-6 level was elevated in 11 of the 22 patients. There were no obvious differences in the clinical data between the two groups, although patients in the normal KL-6 group tended to have a fever. There were no significant differences in the chest X-ray (CXR) score or computed tomography (CT) score between the two groups. The CXR scores were correlated with the serum KL-6 levels. At 1 month after the diagnosis, 11 patients who needed treatment with prednisolone were included in the high KL-6 group. CONCLUSIONS: Patients with normal KL-6 levels showed lower CXR and CT scores. The serum KL-6 level on admission is a useful marker to judge the need for corticosteroid treatment in OP patients.

Familial interstitial pneumonia in an adolescent boy with surfactant protein C gene (Y104H) mutation.

Recent studies have suggested that some cases of familial interstitial pneumonia are associated with mutations in the gene encoding surfactant protein C (SFTPC). We report here a case of familial interstitial pneumonia in an adolescent boy whose paternal grandfather and father suffered from idiopathic interstitial pneumonia (IIP). The patient was asymptomatic but showed an abnormal shadow in the chest at his medical check-up. The surgical biopsy of the patient revealed non-specific interstitial pneumonia and showed pathological findings similar to those in his father's autopsy. Genomic DNA from blood leucocytes of the patient was sequenced for the Thy104His (Y104H) SFTPC mutation. Based on these results, he was diagnosed with SFTPC mutation-associated familial interstitial pneumonia. There has been no clinical, physiologic and radiologic progression for 4 years since the diagnosis. The relation between clinical manifestation and the mutation site of the patient may broaden the spectrum of SFTPC mutation-associated interstitial pneumonia.

Adenovirus-mediated in vivo gene transfer and expression in normal rat liver.

Replication deficient, recombinant adenovirus (Ad) vectors do not require target cell replication for transfer and expression of exogenous genes and thus may be useful for in vivo gene therapy in hepatocytes. In vitro, primary cultures of rat hepatocytes infected with a recombinant Ad containing a human alpha 1-antitrypsin cDNA (Ad-alpha 1AT) synthesized and secreted human alpha 1AT for 4 weeks. In rats, in vivo intraportal administration of a recombinant Ad containing the E. coli lacZ gene, was followed by expression of beta-galactosidase in hepatocytes 3 days after infection. Intraportal infusion of Ad-alpha 1AT produced detectable serum levels of human alpha 1AT for 4 weeks. Thus, targeted gene expression has been achieved in the liver, albeit at low levels, suggesting that adenovirus vectors may be a useful means for in vivo gene therapy in liver disorders.

Asymmetric transcription during post-germinative development of Bacillus subtilis spores.

The relative transcription from L and H strands of Bacillus subtilis DNA during consecutive stages of spore outgrowth was determined and compared to the transcription pattern during log-phase growth of vegetative cells. Pulses of [3H] uridine were administered during early, middle and late outgrowth phases of germination and the RNAs isolated. The asymmetry ratio of H/L as determined by hybridization at saturating RNA/DNA inputs showed a gradual decrease. During the period studied (10-90 and 90-160 min post-induction), about 50 and 35%, respectively, of the radioactive RNA consisted of ribosomal RNA transcripts. The decrease in the H/L asymmetry ratio was due predominantly to the appearance and accumulation of L strand transcripts and not to either changes in the quantity of H strand transcripts nor to fluctuation in the rate of rRNA synthesis.

Asymmetric transcription during post-germinative development of Bacillus subtilis spores. II. Hybrid competition analyses.

Hybrid-competition analyses were done to estimate the relatedness of 3H-labeled mRNA species synthesized during spore germination and log-phase growth. The competitions showed that early in the germination process 10--15 and 1--3% of the RNA transcribed from the H and from the L strand, respectively, were unique and absent during log-phase growth. At later stages, the amounts of the germination-specific H transcripts decreased more rapidly than the L transcripts. The competitions with pulse-labeled log-phase RNAs showed that vegetative genes were transcribed more rapidly from the H strand than from the L strand. Most of the results could be correlated with the observed decrease in the H/L asymmetry ration during spore germination.

Chromosomal organization of rRNA operons in Bacillus subtilis.

Integrative mapping with vectors containing ribosomal DNA sequences were used to complete the mapping of the 10 rRNA gene sets in the endospore forming bacterium Bacillus subtilis. Southern hybridizations allowed the assignment of nine operons to distinct BclI restriction fragments and their genetic locus identified by transductional crosses. Nine of the ten rRNA gene sets are located between 0 and 70 degrees on the genomic map. In the region surrounding cysA14, two sets of closely spaced tandem clusters are present. The first (rrnJ and rrnW) is located between purA16 and cysA14 closely linked to the latter; the second (rrnI, rrnH and rrnG) previously mapped within this area is located between attSPO2 and glpT6. The operons at or near the origin of replication (rrnO,rrnA and rrnJ,rrnW) represent "hot spots" of plasmid insertion.

Efficient generation of autologous peripheral blood-derived cytotoxic T lymphocytes against poorly immunogenic human tumors using recombinant CD80-adenovirus together with interleukin 12 and interleukin 2.

To generate CTLs against poorly immunogenic human tumor cells, we transfected the human CD80 gene into the tumor cells using a replication-deficient adenovirus (Ad) vector. The successful surface expression of CD80 was obtained in both cultured tumor cell lines and primary cultured tumor cells. Transduction of CD80 alone was not sufficient to induce cytotoxicity of peripheral blood lymphocytes against allogeneic tumor cell lines except for melanoma cells. We, therefore, investigated a combined effect of CD80-Ad-infected tumor cells and interleukin 12 (IL-12). Although 7-day cultivation of autologous or allogeneic lymphocytes with CD80-Ad-infected tumor cells and IL-12 slightly enhanced cytotoxicity against some allogeneic tumor cells, no substantial cytotoxicity was observed against autologous tumor cells. When we extended the culture period to 14 days in the presence of IL-2, a prominent enhancement of cytotoxicity was observed against both allogeneic and autologous tumor cells. Cytotoxicity against autologous tumor cells, but not against allogeneic tumor cells, was efficiently inhibited by anti-CD3 monoclonal antibody. Furthermore, the selective cytotoxicity against a panel of targets indicated that the induced CTLs recognize specific antigens on autologous tumor cells. These results suggest that stimulation with a combination of IL-12- and CD80-modified tumor cells and subsequent expansion with IL-2 may efficiently generate tumor-specific CTLs from autologous peripheral blood lymphocytes. Our data imply that the combination of CD80 transduction and suitable cytokines is useful for enhancing antitumor immunity to poorly immunogenic human tumors.

Ex vivo and in vivo gene transfer to the skin using replication-deficient recombinant adenovirus vectors.

The skin has the potential for a variety of gene therapy applications. In addition to local delivery, it is the largest organ of the body, and highly vascular, and thus is an ideal site for systemic delivery of gene products. To evaluate the potential for adenovirus-mediated skin gene transfer, the replication-deficient recombinant adenovirus vectors Ad.RSV beta gal (coding for Escherichia coli beta-galactosidase) and Ad alpha 1AT (coding for human alpha 1-antitrypsin) were used in both ex vivo and in vivo approaches. Following in vitro infection with Ad.RSV beta gal, murine keratinocytes expressed beta-galactosidase. Parallel in vitro studies with Ad alpha 1AT documented de novo synthesis and secretion of human alpha 1AT as shown by [35S]methionine labeling and immunoprecipitation. Quantification of human alpha 1AT in the culture supernatants demonstrated 0.1-0.3 microgram human alpha 1AT secreted/ml-24 h. Evaluation of the serum of mice receiving transplants (10(5) cells/mouse) of Ad alpha 1AT-infected syngeneic keratinocytes demonstrated human alpha 1AT for at least 14 d with maximum levels of 41 ng/ml. To demonstrate the feasibility of direct adenovirus-mediated in vivo transfer of genes to the skin, Ad.RSV beta gal or Ad alpha 1AT were administered subcutaneously to mice. Histologic evaluation after 4 d demonstrated expression of beta-galactosidase in various types of skin cells. Quantification of human alpha 1AT in serum of animals infected subcutaneously with Ad alpha 1AT showed levels of 53 ng/ml at day 4, with human alpha 1AT detectable for at least 14 d. These observations support the feasibility of ex vivo and in vivo gene transfer to the skin mediated by replication-deficient adenovirus vectors.

Adenovirus-mediated gene transfer of glial cell line-derived neurotrophic factor prevents ischemic brain injury after transient middle cerebral artery occlusion in rats.

To examine a possible protective effect of exogenous glial cell line-derived neurotrophic factor (GDNF) gene expression against ischemic brain injury, a replication-defective adenoviral vector containing GDNF gene (Ad-GDNF) was directly injected into the cerebral cortex at 1 day before 90 minutes of transient middle cerebral artery occlusion (MCAO) in rats. 2,3,5-Triphenyltetrazolium chloride staining showed that infarct volume of the Ad-GDNF-injected group at 24 hours after the transient MCAO was significantly smaller than that of vehicle- or Ad-LacZ-treated group. Enzyme-linked immunosorbent assay (ELISA) for immunoreactive GDNF demonstrated that GDNF gene products in the Ad-GDNF-injected group were higher than those of vehicle-treated group at 24 hours after transient MCAO. Immunoreactive GDNF staining was obviously detected in the cortex around the needle track just before or 24 hours after MCAO in the Ad-GDNF group, whereas no or slight GDNF staining was detected in the vehicle group. The numbers of TUNEL, immunoreactive caspase-3, and cytochrome c-positive neurons induced in the ipsilateral cerebral cortex at 24 hours after transient MCAO were markedly reduced by the Ad-GDNF group. These results suggest that the successful exogenous GDNF gene transfer ameliorates ischemic brain injury after transient MCAO in association with the reduction of apoptotic signals.

Nucleotide sequence of a member of the chicken H2B histone-encoding gene family.

The nucleotide sequence of a gene from the chicken H2B histone-encoding gene family has been determined. Our findings, together with those in a previous paper [Grandy and Dodgson, Nucleic Acids Res. 15 (1987) 1063-1080], show that the seven H2B genes encode three different histone variants.

Family functioning, temperament, and psychologic adaptation in children with congenital or acquired limb deficiencies.

Family functioning and child temperament variables were investigated as predictors of psychologic and social adaptation in 42 children with congenital or acquired limb deficiencies. Higher psychologic and social adaptation were seen when there was more family cohesion and moral-religious emphasis and organization, in combination with less family conflict. With regard to child temperament, more emotionality predicted greater internalizing and externalizing behavior problems and less social competence. In addition to the main effects of the family functioning and child temperament predictor variables, the interaction between family cohesion and child emotionality significantly predicted both internalizing and externalizing behavior problems. The findings are discussed in terms of the risk and protective effects of family functioning domains and temperament on the psychologic and social adaptation of children with visible physical handicaps.

Soluble interleukin-2 receptor in sera of patients with pulmonary tuberculosis.

Interleukin-2 receptor, the complex of IL-2R-alpha and/or IL-2R-beta, is expressed mainly on T-lymphocytes, and the soluble form of IL-2R-alpha (sIL-2R-alpha) has been reported to be detected in the serum of patients with lymphoproliferative disorders or disease characterized by the cellular immune reaction. We measured serum sIL-2R-alpha levels among patients with pulmonary diseases and found that sIL-2R-alpha levels were significantly elevated in patients with active pulmonary tuberculosis (1,327 +/- 209 U/ml) and sarcoidosis (1,037 +/- 115 U/ml) when compared with healthy volunteers (468 +/- 49 U/ml, p less than 0.01). Among patients with pulmonary tuberculosis, the sIL-2R-alpha levels were high in sera from patients with extensive parenchymal lesions on the roentgenogram (2,745 +/- 705 U/ml) and patients with tuberculous pleurisy (2,111 +/- 679 U/ml). In contrast, the sIL-2R-alpha levels in tuberculous patients with minimal lesion (455 +/- 92 U/ml) or moderate lesion (1,082 +/- 189 U/ml) were not significantly elevated when compared with healthy volunteers. After the treatment with antituberculosis agents, serum sIL-2R-alpha levels decreased in accordance with improvement of roentgenographic findings and laboratory data. These results suggest that serum sIL-2R-alpha level may be useful as a monitor for the disease activity in patients with pulmonary tuberculosis.

Adenovirus-mediated glial cell line-derived neurotrophic factor gene delivery reduces motor neuron injury after transient spinal cord ischemia in rabbits.

OBJECTIVE: Glial cell line-derived neurotrophic factor (GDNF) has protective effects on various injuries involving the central and peripheral nervous systems in vitro and vivo. However, the possible protective effect of GDNF on spinal cord ischemia and the exact mechanism involved in the ameliorative effect of GDNF on ischemic spinal cord injuries are not fully understood. Therefore, we investigated the possible protective effect of the adenovirus-mediated GDNF gene delivery on transient spinal cord ischemia in rabbits. METHODS: The adenoviral vector (lacZ gene as a control or GDNF gene contained) was injected directly into the lumbar spinal cord via a needle inserted into the dorsal spine 2 days before the animal was subjected to 15 minutes of spinal cord ischemia induced by infrarenal aortic occlusion. In situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL staining) was performed, and temporal profiles of the GDNF and caspase-3 (caspase-3 is the marker of apoptotic change) immunoreactivity were investigated. RESULTS: In the control rabbit, the majority of motor neurons showed selective cell death at 7 days of reperfusion. Immunocytochemistry showed that in situ TUNEL staining was selectively detected at 2 days of reperfusion in motor neuron nuclei. GDNF and caspase-3 were selectively induced in the motor neuron cells at 8 hours of reperfusion. In the GDNF-treated group, a large population of motor neuron cells was still surviving at 7 days after having been subjected to 15 minutes of ischemia. Unlike the control group, the GDNF-treated group expressed GDNF persistently. Induction of TUNEL staining and immunoreactivity for caspase-3 were greatly reduced by the GDNF treatment. CONCLUSION: These results suggest that the reduction in motor neuron death by GDNF was greatly associated with a reduction in DNA fragmentation and apoptotic signals of the caspase-3 cascade; they further suggest a great potential for gene therapy for paraplegic patients in the future.

Intrathecal injection of high-dose meglumine amidotrizoate with complete recovery.

Rarely we are faced with accidental spinal injection of potentially toxic substances. We present 2 cases in which amidetrizoate, water-soluble ionic contrast medium, was accidentally injected intrathecally. Our treatment consisted of vigorous hydration and barbiturate coma. This report suggests that for water-soluble ionic contrast media increasing cerebrospinal fluid circulation by vigorous hydration may be as effective as spinal lavage in diminishing toxicity.

Serum lipoprotein(a) levels before and after subtotal thyroidectomy in subjects with hyperthyroidism.

Lipoprotein(a) [Lp(a)], a lipoprotein that structurally resembles low-density lipoprotein (LDL), contains apolipoprotein(a) [apo(a)] and apolipoprotein B-100 (apo B). There exists a close inverse correlation between serum concentrations of LDL or apo B and concentrations of thyroid hormone in patients with thyroid disease, probably due to a change in LDL receptor activity. To clarify the relations between thyroid hormone and Lp(a), we measured serum Lp(a) levels in 13 hyperthyroid subjects before treatment (stage H), during the euthyroid stage induced immediately before performing a subtotal thyroidectomy (stage E), and during the hypothyroid stage observed transiently after the operation (stage L). The mean serum concentration of Lp(a) increased significantly (P = .01) from 9.4 mg/dL in stage H to 26.8 in stage L through the level of 15.5 mg/dL in stage E. There was no significant difference between the mean serum concentration of Lp(a) in these patients in stage E and healthy controls (14.2 mg/dL). There was a low but statistically significant negative correlation between the Lp(a) level and the serum free thyroxine (fT4) concentration (r = .31, P < .05). The results suggest that thyroid hormone is a potent modulator of Lp(a) metabolism.

Neuron-targeted gene transfer by adenovirus carrying neural-restrictive silencer element.

Adenovirus transfers genes to a wide range of cell types, but its application to neurons has been hampered by its reduced efficiency of infection as compared with that for glia. To achieve neuron-targeted gene transfer, we have produced an adenovirus carrying the reporter lacZ gene driven by the SCG10 minimum promoter containing the neural-restrictive silencer element (NRSE), which element selectively represses the transcription of genes in non-neuronal cells. When rat hippocampal slice cultures were infected with NRSE-bearing adenovirus, beta-galactosidase-positive cells were mostly pyramidal and granular neurons, whereas infection with virus carrying a mutated NRSE resulted in beta-galactosidase expression in both neurons and glia. The results suggest that the adenovirus carrying NRSE to be a useful tool for neurontargeted gene transfer.

In vivo adenovirus-mediated gene transfer and the expression in ischemic and reperfused rat brain.

In an attempt to study whether ischemic brain could express a foreign gene in vivo, a replication-defective adenoviral vector containing the Escherichia coli lacZ gene was directly injected into the ischemic or reperfused cerebral cortex of rat, and temporal and spatial profiles of the exogenous gene expression were compared with that of the control brain. Right middle cerebral artery (MCA) of rat was continuously occluded by an insertion of nylon thread for 2 days, or only transiently occluded for 90 min and then the blood flow was restored for 21 days. The adenoviral vector was administered just after the MCA occlusion or reperfusion in the case of continuous ischemia and reperfusion, respectively. Adenoviral vector was transferred into the continuous ischemic brain, and the lacZ gene was expressed until 2 days of the occlusion in the cerebral cortex of the occluded MCA territory with the number of expressing cells smaller and the staining just weaker than that of the control brain. In contrast, expression of the lacZ gene was not or only minimally observed in the reperfused brain until 2 days. However, the expression dramatically exploded at 7 days of reperfusion at a level similar to that of the control, and the expression diminished by 21 days. A few neurons in the ipsilateral thalamus, hypothalamus, and basal ganglia, and in the contralateral cerebral cortex expressed the lacZ gene at 7 days after reperfusion, a phenomenon similar to the case of the control. The majority of brain cells that expressed the lacZ gene were neurons, and a part (5-10%) were astroglial cells. Traumatic injury and immunological response in the brain were minimal both in the cases of control and ischemia/reperfusion. The present study shows an effective gene transfer and the expression in neural cells of ischemic and reperfused brains in vivo, and suggests a great potential of the gene therapy for ischemic stroke patients in the future.

Dissociative expression of adenoviral-mediated E. coli LacZ gene between ischemic and reperfused rat brains.

A replication-defective adenoviral vector containing the E. coli lacZ gene was directly injected into the ischemic or reperfused cerebral cortex of rats. An administration of adenoviral vector showed a slight to moderate expression of the lacZ gene in the cerebral cortex of the middle cerebral artery (MCA) region until 2 days after the MCA occlusion. In contrast, expression of the lacZ gene was not observed, or only minimally so, in the reperfused brain until 2 days after a 90 min of transient MCA occlusion. However, the lacZ expression dramatically increased at 7 days after the reperfusion, then diminished by 21 days. The majority of brain cells that expressed the lacZ gene were neurons and a fraction (5-10%) were astroglial cells. The present study showed that an exogenous gene was transferred and expressed in neural cells of ischemic and reperfused brains in vivo, but the temporal profile of the expression is dissociative.

Expression of adenovirus-mediated E. coli lacZ gene in skeletal muscles and spinal motor neurons of transgenic mice with a mutant superoxide dismutase gene.

A replication-defective recombinant adenoviral vector containing E. coli lacZ gene was injected into the right biceps brachii muscles of transgenic mice carrying mutant human Cu/Zn superoxide dismutase (SOD1) gene and non-transgenic wild-type mice at 27 weeks of age. Although the transgenic mice showed remarkable neurogenic muscular changes and a marked motor neuron loss in the anterior horn of spinal cord, the lacZ gene was widely expressed in all the injected muscles of transgenic mice as well as of wild-type mice at 7 days after the injection. In one transgenic and two wild-type mice, the lacZ gene expression was first detected in a few motor neurons of right lower cervical cord (C5-C6). These results demonstrate that an adenovirus-mediated foreign gene is transferred and expressed in skeletal muscles both of normal and transgenic mice model for familial amyotrophic lateral sclerosis (FALS), and also, in the spinal motor neurons, may be transferred by retrograde transport from innervated muscles.