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Paper chromatographic procedures may be used to detect cephalothin and its metabolite desacetylcephalothin in urine, plasma, synovial fluid, and cerebrospinal fluid. Protein-bound antibiotics are released from plasma, cerebrospinal fluid, and synovial fluid by dilution with an equal volume of dimethylformamide. Data are presented on the sample preparation, paper chromatographic system, and other specific techniques.

For the purpose to reveal the characteris6cs of the synovial fluid of the chronic rheumatoid arthritis the proteins of the synovial fluid and blood serum have been analysed by employing the methods of electrophoresis, gel filtration on Sephadex G-200 column and ultracentrifugation. Waaler-Rose test and latex fixation test have also been made on each protein fraction, and the following results were obtained. 1) The total protein level of synovial fluid, which is 3/5 of that of the serum, is slightly higher than that of control. 2) Fractionation of the synovial proteins by electrophoresis revealed nearly the same protein contents in each fraction in percentage as that of comparable fraction of the serum protein, with a slight increase in γ-globulin fraction. 3) The fractionation by Sephadex column G-200 give three peaks both in serum and synovial fluid, 19 S, 7Sand 4S. 4) 19S fraction of the synovial fluid, which is mainly of γ-globulin, showed a higher level than that of the synovial fluid from the controls. 5) Rheumatoid tests gave positive reaction in the 1st peak containing 19S γ-globulin from the synovial fluid and blood serum.

Immunological investigation of four patients with meningococcal meningitis who developed arthritis or cutaneous lesions showed circulating meningococcal antigen at the time of presentation in each patient. It was cleared from the circulation over the next few days. Circulating antibody was detectable in three of the four patients about a week after the onset of the illness. A marked fall in the serum C(3) level occurred in two patients at about that time. Deposits of meningococcal antigen, immunoglobulin, and C(3) were detected in the synovial fluid white cells of the two patients studied and in one of three skin biopsies examined. These findings suggest that the arthritis and cutaneous lesions of meningococcal meningitis may be due to immune complex formation.

Out of 717 patients with meningococcal disease 53 showed one or more of the three allergic complications: 47 (6·6%) developed arthritis, 12 (1·7%) developed cutaneous vasculitis, and 6 developed episcleritis. These complications, which were often multiple, occurred six to nine days after the beginning of the illness and three to six days after the start of successful antibiotic therapy. Those patients with severe systemic disease were prone to the complications. Histological and bacteriological study of the arthritis and vasculitis showed that these lesions were probably not due to persisting infection and suggested that they might be due to immune complex disease.

Normal, postmortem, traumatic and rheumatoid synovial fluids were studied by paper electrophoresis, ultracentrifugation and starch gel electrophoresis. It could be demonstrated that the change in the α1/α2 globulin ratio, particularly prominent in normal and postmortem synovial fluids as compared with normal serum, is a consequence of the greatly increased relative concentration of α1-acid glycoprotein and a significant decrease in the relative concentration of the haptoglobins and the 19S α2-macroglobulin. Specimens de liquido synovial—normal, post morte, traumatic, e rheumatoidee—esseva studiate per electrophorese a papiro, ultracentrifugation, e electrophorese a gel de amylo. Il esseva possibile demonstrar que le deviation del proportion globulina α1 a globulina α2 ab le valor trovate in le sero—un deviation particularmente marcate in liquido synovial normal e post morte—es un consequentia del grandemente augmentate concentration relative de glycoproteina acide α1 e del significativemente reducite concentration relative del haptoglobinas e del macroglobulina α2 19S.

Biochemical studies concentrating on a paper electrophoretical analysis were made with the following results on the synovial fluid in an attempt to make a diagnosis of arthropathy.1. The amount of protein was greater than that in a control group and was proportional to the intensity of the inflammation.2. The severer the inflammation was, the lower was the viscosity of the synovial fluid. The viscosity indicated the local findings well.3. The flocking number reaction (Takata) and particularly the mat delayed reaction were of value in observing the course of diseases.4. Although the Tokata reaction, the Chinin reaction and the cadmium reaction did not occur in the synovial fluid the clinical significance of the Weltmann's coagulation band for serum was recognized.5. A marked increase in mucoprotein took place in patients blood serum with rheumatosis. This finding was of significance in following the course of the diseases.6. No changes between the types of disease took place in alkali phosphatase and in inorganic substances Na, K, P and Ca.7. Amarked decrease of A/G ratio was observed in rheumatoid arthritis which was proportional to the intensity of inflammation.

During thepast4years allsynovial fluids taken forexamination havebeensubmitted toelectrophoresis onpaperforestimation ofthedifferent protein fractions. A sufficient numberhasnow beenexamined toallow amoredetailed analysis of theresults thanwaspossible inourearlier report (Wilkinson andJones, 1962). Material andMethods Paper electrophoretic studies weremadeonserumand synovial fluid fromoneorbothknees ofpatients with thediagnoses listed inTableI,using themethods previously described (Wilkinson andJones, 1962). Thediagnoses wereestablished fromtheclinical and radiological features, butinaddition successful needle biopsies wereavailable frommorethanhalf thepatients asindicated inTable I.Inorder that thehistological features couldbecorrelated withthesynovial fluid changes, eachbiopsy wasstudied byoneobserver (B.S.J.) without knowledge oftheclinical orbiochemical details. Particular attention waspaid tothedegree of synoviocyte hyperplasia, vascularity, lymphocyte infiltration, andplasma cell infiltration, andeachofthese features wasgraded +, ++,or+++,tofacilitate comparisons. TABLEI PATIENTSSTUDIEDAND SPECIMENSANALYSED

The mononuclear cells from the joint fluid of certain patients with inflammatory arthritis are shown to be more cytotoxic to a polyploid strain of homologous liver cells than were equivalent numbers of peripheral blood lymphocytes from the same donors. The diagnostic categories of these patients are essentially similar to those of a group of patients studied by Hedberg (1967) in whom joint fluid mononuclear cells were cytotoxic to monolayers of human foetal fibroblasts. Evidence is presented to show that this cytotoxic activity is attributable to lymphocytes, as opposed to polymorphonuclear leucocytes or phagocytic mononuclear cells which adhere to glass. It is not excluded that the latter population may also have some cytotoxic potential. It is argued that the target cell damage described in this paper is probably not induced by any antigen on the target cell. The implications of this suggestion are discussed in relation to the pathogenesis of inflammatory joint disease.