Chiyoko N. Inoue

Minocycline-related lupus in childhood

Sir: Systemic lupus erythematosus (SLE) is a connective tissue disease whose specific cause or causes remain undetermined. However, drugs and other agents can produce a lupus-like syndrome, drug-related lupus [6]. We have cared for a 14-year-old girt with lupus-like syndrome caused by prolonged minocycline therapy for acne. A 14-year-old girl was admitted because of spiking fever for 8 days and arthralgia in the right elbow. Two months before admission, she had begun to receive minocycline 200 mg daily for facial acne. There was no history of hair loss or photosensitivity. Chest X-ray was normal. Laboratory data were as follows: Hb 11.6 g/dl, WBC 2900/mm 3, ESR 50 mm/h, serum GPT 284 IU/1 and CRP 0.9 mg/dl. IgG, A, M, C3, C4 and CH50 values were normal. Serum antinuclear antibody (ANA) was positive at 1 : 640 with a nucleolar pattern. Anti-DNA, anti-Sm, anti-histone antibodies and LE test were all negative. Administration of piperacillin was ineffective. On the 6th hospital day, her physician realized that she had no discontinued minocycline, and ordered that it be stopped. After the cessation of minocycline, she defervesced rapidly and the arthralgia disappeared. She was discharged from the hospital after 21 days. At home, she had a recurrence of fever and arthralgia when she took minocycline for 2 days. During follow-up for 8 months, the GTP level and ANA titre gradually normalized and the patient was asymptomatic without any special treatment. HLA study revealed that she had HLA-DR2, which is thought to be the common HLA antigen in Japanese and Caucasian patients with SLE [2]. Agents causing drug-related lupus have been classified into three groups: (1) those with definite proof of association, (2) those with possible association, and (3) those with unproved association. Tetracyclines fall into the third group [6]. The first possible case of drug-related lupus with minocycline was described by Matsuura et al. [5] in 1993. In this report, a 22-year-old Japanese woman showed lupus syndrome following administration of minocycline for 2 years for the treatment of acne [5]. The case of Matsuura and ours are very similar: both involved young woman with long histories of minocycline therapy for acne who possessed HLA-DR2, which is a marker for SLE. As ache therapy, tetracyclines are often administered for several months to a few years until the desired clinical improvement is evident. In addition to antimicrobial activity, tetracyclines have a number of anti-inflammatory actions such as inhibition of leucocyte chemotaxis [1] and enhancement of interleukin1 f3 release [31. Therefore, it is reasonable to assume that tetracyclines interact with host cell-mediated immune defenses, which may contribute to their efficacy in the treatment of inflammatory lesions, and that chronic exposure to tetracyclines could perturb the host immune system, resulting in autoimmune disorders. Furthermore, acne inflammation mainly involves T-cell mediated immunity [4], suggesting that acne may be a co-factor in the pathogenesis of minocycline-related lupus. We believe that the prolonged administration of minocycline to individuals who have a hereditary predisposition to develop SLE may induce lupus-like syndrome. We feel that physicians should be aware of this possibility, especially in the treatment of acne.

RECONSTRUCTION OF TUBULAR STRUCTURES IN THREE-DIMENSIONAL COLLAGEN GEL CULTURE USING PROXIMAL TUBULAR EPITHELIAL CELLS VOIDED IN HUMAN URINE

SummaryHuman proximal tubular (PT) epithelial cells were isolated from urine and monoclonally cultured as monolayers for 1 wk, after which they were subcultured between two layers of collagen gel, designated a “collagen gel sandwich.” Under these culture conditions, PT cells formed three-dimensional tubular structures exhibiting distinct polarized cell morphology. Scanning and transmission electron microscopic studies showed that they bore numerous microvilli at the apical surface and that they closely contacted the collagen gel at the basal surface. These studies indicate that PT cells exfoliated in urine still exhibit the potential to proliferate and form organized structures mimicking in vivo tubules. Because of the current lack of useful culture systems for human tubular epithelial cells originating from kidney tissue, we suggest that this unique culture system using voided PT cells in urine could open up new avenues to study not only the mechanisms of morphogenesis but also the physiology of human PT cells.

Nucleotide sequence determination of mouse, chicken and Xenopus laevis rig cDNAs: the rig-encoded protein is extremely conserved during vertebrate evolution.

Mouse, chicken and Xenopus laevis homologues to rig (rat insulinoma gene) cDNA were isolated and their nucleotide sequences were determined. Each homologue encoded a 145-amino acid protein; the amino acid sequence remained invariant in the murine and avian genes, and there were only 6 amino acid substitutions in the salientian gene. The evolutionary rate calculated for rig mRNA was sufficiently low to be viewed as evidence that rig is vital to vertebrate species. Southern blot analysis indicated that haploid sets of the mammalian genomes contain several copies of rig or rig-related sequences, whereas there appeared to be only one copy in the amphibian and bird genomes. The possibility that rig belongs to the class of housekeeping genes is discussed.

Bimodal effects of platelet-derived growth factor on rat mesangial cell proliferation and death, and the role of lysophosphatidic acid in cell survival

Although mesangial cell death has been shown to be correlated with mesangial cell mitosis in vivo, little is known about how these two apparently opposite events are regulated. We show that the addition of platelet-derived growth factor (PDGF; 10-50 ng/ml) to primary cultured rat mesangial cells for 24 h caused continuous proliferation along with simultaneous cell death. This process was accompanied by the fragmentation of DNA into nucleosomal oligomers, the development of apoptotic morphological changes in the nucleus, and increased expression of p53. Accumulation of lactate dehydrogenase (LDH) was also observed in the culture medium, suggesting that both apoptosis and necrosis are involved in the cell death mechanisms observed. We also observed that addition of 30 microM lysophosphatidic acid (LPA) to the culture medium greatly suppressed PDGF-induced cell death, leading to synergistically enhanced mesangial cell proliferation. DNA fragmentation, p53 expression and LDH release were all suppressed by LPA. We suggest that PDGF is a bifunctional molecule in mesangial cells that evokes both cell proliferation and cell death simultaneously, whereas LPA is a survival factor. We speculate that PDGF and LPA may play important roles in the progression or exacerbation of proliferative glomerulonephritis.

Expression of the insulinoma gene rig during liver regeneration and in primary cultured hepatocytes

We have isolated a novel gene, rig (rat insulinoma gene) from rat insulinomas. In the present study, rig was found to be expressed in rat regenerating liver and in primary cultured rat hepatocytes. The level of rig mRNA was increased at the proliferative phase of liver regeneration. In synchronously cultured hepatocytes, the rig mRNA level was elevated at the G1 phase of the cell cycle and the rig-protein was accumulated in the nuclei during the S phase. These results indicated that rig could be involved in a more general way in growth or cell replication.

Mitogenic action of lysophosphatidic acid in proximal tubular epithelial cells obtained from voided human urine

Focal tubular cell multiplication at sites on an injured nephron is a critical event in the recovery phase following acute tubular necrosis. During this process, numerous viable tubular cells exfoliate and are shed into the urine. Lysophosphatidic acid (LPA) is generated in the plasma membrane of injured cells and acts as an intercellular mediator of various biological processes, including inflammation, proliferation and repair. In the present study, exfoliated proximal tubule (PT) cells were isolated from human urine and the mitogenic effects of LPA were investigated as a model of repair and proliferation following renal injury. LPA stimulated a 23. 5% increase in DNA synthesis, a 29.4% increase in cell number and an 86.6% decrease in cAMP content. All of these responses were pertussis toxin sensitive, indicating the involvement of G(i)-type G-proteins in LPA signalling. Conversely, the LPA-induced DNA synthesis and the decrease in intracellular cAMP content were insensitive to wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K), suggesting a mitogenic response via PI3K-independent mechanisms. Furthermore, we detected specific mRNA transcripts for the recently cloned human LPA-receptors, endothelial differentiation gene (Edg)-2 and Edg-4 (Edg-2>>Edg-4) by reverse transcription-PCR in PT cells. Our data suggest that LPA may behave as a local growth factor in PT cells following tubular injury.

Steroid-sparing effect of mizoribine in long-term nephrotic syndrome of children.

were reported in a collaborative study of 661 patients [5-7] . In conclusion, three patients, in whom Hodgkins disease followed steroid-resistant idiopathic nephrotic syndrome treated with cyclosporine A, have been observed. Despite the well-known relationship between these diseases, the possibility that cyclosporine A could induce Hodgkins disease in patients with idiopathic nephrotic syndrome must be considered.

A new approach to mRNA in proximal tubule cells of patients with CLCN5 channelopathy

Abstract ClC-5 is a chloride channel whose gene mutations have been reported to be associated with X-linked nephrolithiasis (XRN), X-linked recessive hypophosphatemic rickets (XLRH), Dent disease, and idiopathic low-molecular-weight proteinuria (ILMWP) in Japanese children. To establish more efficient screening for CLCN5 abnormalities, we developed a new diagnostic method using reverse transcription and polymerase chain reaction (RT-PCR) of cultured renal tubular cells from the urine of patients. Using this new method, we successfully detected microdeletion of ClC-5 mRNA in a patient and splicing abnormality of the CLCN5 Cl channel.

Possible Mechanisms Underlying Epipharyngeal Abrasive Therapy (EAT) with ZnCl 2 Solution for the Treatment of Autoimmune Diseases and Functional Somatic Syndrome

Chronic epipharyngitis is a common latent but serious condition that may contribute to a wide range of diseases in humans, including collagen diseases, glomerulonephritis and autonomic nervous disorders. In a previous study, we presented a putative causal role of chronic epipharyngitis in the development of functional somatic symptoms and syndromes following human papillomavirus vaccination by demonstrating a significant improvement in symptoms following abrasive therapy using ZnCl2 on the epipharynx. Since this initial study, we have expanded our clinical experience, providing epipharyngeal abrasive therapy to 988 patients with confirmed chronic epipharyngitis associated with a wide variety of clinical symptoms. These symptoms could be classified into three broad categories, namely local inflammation-referred, autoimmune-related, and neuroendocrine symptoms. Symptom alleviation was achieved in the majority of patients with repeated epipharyngeal abrasive therapy.Through an in-depth review of the literature on epipharyngeal abrasive therapy, combined with our clinical experience, we propose three mechanisms underlying the therapeutic effects of epipharyngeal abrasive therapy: the astringent anti-inflammatory effect of ZnCl2, a blood-letting effect that promotes removal of epipharyngeal activated lymphocytes and drainage of excess inflammatory fluids containing various antigens, cytokines or noxious substances, and a neuromodulation effect achieved through stimulation of the vagus nerve. These effects can be explained within the context of current understanding of immunology, lymphology and neuroscience.Our hypothesis-driven review provides a theoretical basis for the observed therapeutic effects of epipharyngeal abrasive therapy in ameliorating various diseases, including functional somatic symptoms and syndromes following human papillomavirus vaccination.