Purification and characterization of a novel human red cell enzyme with diphenol oxidase activity.

A new enzyme protein, diphenol oxidase, has been isolated and purified from human red cells and catalyzes the in vitro formation of adrenochrome from epinephrine and of melanin from dihydroxyphenylalanine. Some immunological and physicochemical properties of this protein have been studied.

Rectal swab testing before prostate biopsy: experience in a VA Medical Center urology practice.

BACKGROUND: Infectious complications following transrectal ultrasound prostate biopsy (TRUSBx) have been increasing. Pre-biopsy prophylaxis with fluoroquinolone (FQL) antibiotics is a recommended and accepted practice. Increasing emergence of FQL-resistant bacteria is believed to be related to the increase in infectious complications. We sought to determine the effect of targeted antibiotic prophylaxis (TAP) before TRUSBx on infectious complications in our practice. METHODS: TAP was introduced in our practice in 2012. A retrospective analysis was performed analyzing infectious complications from TRUSBx before and after TAP was introduced. Two hundred forty-four patients underwent TRUSBx with TAP directed by bacterial antibiotic sensitivity identified on rectal swab. A group of two hundred sixty-four consecutive patients who underwent TRUSBx in our practice before introduction of TAP were chosen for comparison. Infectious complications were recorded and compared between groups. Prostate volume, PSA, number of biopsy cores, finding of prostate cancer, presence of diabetes, race and age were also compared. RESULTS: The infectious complication rate after TRUSBx in the pre-TAP group was 7/264 (2.65%), the rate in the TAP group was 1/244 (0.41%), a statistically significant difference (P=<0.05). There were no differences between groups in regards to prostate volume, number of prostate biopsy cores, race and presence of diabetes. The rectal swab group was younger (65.4 ± 6.0) than the non-swab group (67.9 ± 6.2), had higher PSA values, and a higher chance of prostate cancer on biopsy. CONCLUSIONS: The use of TAP based on rectal swab testing significantly lowered our infectious complication rate for TRUSBx. TAP is now adopted as standard practice before TRUSBx in our center. The younger age and higher chance of prostate cancer on biopsy in the rectal swab group, we believe, is due to implementation of recent guidelines directing urologists be more selective in recommending prostate biopsy to older men.

[Diphenoloxydase (DPOx) in red cell membranes]. / Etude des diphénoloxydases (DPOx) dans les membranes de globules rouges.

Two diphenoloxydases (DPOx) with similar molecular weights (158 000) are found in normal human red cell hemolysates. One of these enzymes appears to be tightly bound to the membranes while the other is not, or at most only weakly bound.

Progressive muscular dystrophy. Functional improvement after a renal allograft.

From his first years a child showed signs of a primary and rapidly developing muscular dystrophy. The diagnosis was established by an increased serum CK level and by electromyography and muscle biopsies. Afterwards this child developed a severe renal deficiency which needed binephrectomy and the graft of a normal kidney. During the few months just after the graft, the disability increased and the patient could not stand upright by himself. Later on, he gradually became able to walk on his own and without bracing. He could climb stairs and stand up from the floor. The CK activity returned to normal. At present, 4 years after the graft (the patient is 16 years), the improvement of his functional abilities is constant, although the CK activity has increased again. In this article we give evidence that this patient suffers from a primary muscular dystrophy. We discuss the type of dystrophy concerned. We believe that it is the graft of a normal kidney which was responsible for the improvement observed, and not the physiotherapy or the drugs administered after the graft.

[Distribution of diphenoloxidases (DPOx) in various tissues of various animal species (author's transl)]. / Distribution des diphenoloxydases (DPOx) dans divers tissus de diverses especes animales

An oxidative activity has been demonstrated in different tissues of various animals species (man, rabbit, hen, rat, mouse) measuring it by means of adrenaline, by starch gel electrophoresis and acrylamide gel electrofocusing, followed by incubation of these gel with DOPA. Kidney, liver and erythrocytes of rabbit, rat, mouse and hen, human erythrocytes and platelets display a high oxidative activity. This activity, which is not completely inhibited by KCN, is found in one band on starch gel electrophoresis and in two distinct bands on electrofocusing in the case of platelets and erythrocytes of man, rabbit and rat. The starch gel electrophoretic migration and the electrofocusing pattern are tissue and species dependent.

[Diphenoloxidases: presence in the membrane of human erythrocyte (author's transl)]. / Diphénoloxydases: présence dans les membranes de globules rouges

Diphenoloxidases, enzymes which accelerate the auto-oxidation of epinephrine and Dopa, have been described by one of us in blood platelets. Earlier we identified these enzymes in different animal species and particularly in human red blood cells. With the object of localising these enzymes and of understanding their function in vivo, we separated the ghosts of red blood cells according to the method described by Fairbanks G., Steck, T.L. and Wallach, D.F.H. (1971) (Biochemistry 1, 2606) and using the protease inhibitors diisopropylfuorophosphate (DFP) (10(-3) M) and 6-aminocaproic acid 10(-2) M in sodium phosphate buffer, 5 X 10(-3) M (pH 8). These ghosts, totally free of haemoglobin, were first of all pulverised in liquid nitrogen then treated ultrasonically. The supernant shows the presence of a band of diphenoloxidase activity on starch gel electrophoresis and two bands on isoelectrofocusing in polyacrylamide gel pH 5 to pH 8 after incubation with 0.02 M Dopa, 0.076 M Tris, 0.005 M citric acid, 0.004 M magnesium, pH 8.7 for 2 h at 37 degrees C. These enzymes differ from (Na, K)-ATPase in that they are neither inhibited by DFP (10(-1) M) nor by EDTA (10(-2) M) but are inhibited by lead acetate 10(-2) M. Like (Na, K)-ATPase diphenoloxidases are present at membranes level. The role in vivo of these diphenoloxidases in ATPase activity of red blood cells is discussed.

[Change in heart function in Duchênne de Boulogne myopathies at the severe progressive stage]. / Altération de la fonction cardiaque dans les myopathies de Duchênne de Boulogne au stade évolutif sévère.

Left ventricular function was studied in 15 patients with rapidly progressive Duchenne de Boulogne muscular dystrophy. Only one patient had a previous history of cardiac failure. All patients had typical electrocardiographical changes. Haemodynamic and angiographic investigation was performed in all patients. Three groups of patients were distinguished according to angiographic criteria : group I comprising 7 normal patients. group II comprising 4 "intermediary" patients, and group III of 4 patients with hypokinetic cardiomyopathy. Segmental ventricular dyskinesis was observed in 6 patients. Mitral incompetence was present in 2 cases and mitral valve prolpse in 1 case. All patients had a normal resting cardiac index. Patients in group II and III had end diastolic left ventricular pressures greater than 15 mmHg. An intravenous angiotensin test was performed in 4 patients from groups I and II. Impaired left ventricular function was demonstrated in patients thought to be normal in the basal state. Finally, myocardial involvement in this group of patients runs a parallel course to the duration and severity of the peripheral muscular disease.

[Results of the treatment of Duchenne de Boulogne myopathy by early physiotherapy. A comparative study with untreated subjects]. / Résultats du traitement de la myopathie de Duchenne de Boulogne par physiothérapie précoce. Etude comparative avec des sujets non traités.

163 patients presenting with Duchenne Muscular Dystrophy (DMD) were treated with daily long-term physiotherapy in two specialized centers. This treatment was undertaken at a time when patients were still able to walk alone. II was continued in the centers up to an age when patients could no longer walk alone. As an average, the latter age was delayed from 3 to 4 years (according to the precocity of treatment) as compared with untreated patients. The comparisons between treated and untreated siblings established the significance of the results obtained. These results may be favourably compared with those obtained with other methods. The reduction of the progression of the disease at the level of the pelvic girdle mostly accounts for the gain.

A simplified, improved techniqe for the Collis gastroplasty for dilatable esophageal strictures.

A safe, simple, effective and fast gastrophasty for the dilatable strictured esophagus has proved sound in dogs and feasible and effective in twenty-five patients. Several of the patients who had been operated upon elsewhere had recurrences. Postoperative follow-up studies of the 25 patients operated upon have revealed freedom from substernal pain, dysphagia, aspiration and bleeding in the upper part of the gastrointestinal tract. Encouraged by the success of the surgical techinique in patients with stricture of the esophagus, we have recently applied the same technique in the repair of hiatal hernia without stricture.

Diphenoloxidases in various forms of myopathy which are transmitted by different genetic mechanisms.

In a previous article (Demos et al. 1981), we reported a significant and specific reduction of the activity index (AI) of the diphenoloxidases (DPox) in patients and heterozygotes with progressive Duchenne muscular dystrophy (DMD), which is transmitted genetically by female subjects by a sex-linked recessive mechanism (SLR). This same anomaly was detected in patients suffering from other types of dystrophy: Becker, limbgirdle, fascio-scapulo-humeral, and in heterozygotes of either sex in diseases transmitted by an obviously recessive autosomic mechanism. These anomalies were detected using blood spots collected on absorbent paper and stored at 4 degrees C for different periods. They were of the same type as had previously been detected using blood platelets (Demos 1973).

Diphenoloxidases in X-linked recessive (Duchenne) muscular dystrophy.

In extracts derived from whole blood, a high molecular weight fraction of the diphenoloxidase enzymes has a significantly diminished specific activity in patients and definite carriers (heterozygotes) of the X-linked, recessive (Duchenne) form of muscular dystrophy. This anomaly was studied using spots of blood which had been collected on absorbent paper and stored at 4 degrees C for variable periods of time. Fractions enriched in the enzymes were obtained by subjecting aqueous extracts of the spots to treatment with an anion exchange resin (DEAE Sephadex A 50) followed by gel filtration on Sephadex G-25. It is of interest that this anomaly was observed in some definite carriers of the mutant gene who had on several occasions a serum creatine kinase level in the normal range. The significance of these observations is discussed.