Bacteriophage Tail-Spike Proteins Enable Detection of Pseudaminic-Acid-Coated Pathogenic Bacteria and Guide the Development of Antiglycan Antibodies with Cross-Species Antibacterial Activity.

Pseudaminic acid (Pse), a unique carbohydrate in surface-associated glycans of pathogenic bacteria, has pivotal roles in virulence. Owing to its significant antigenicity and absence in mammals, Pse is considered an attractive target for vaccination or antibody-based therapies against bacterial infections. However, a specific and universal probe for Pse, which could also be used in immunotherapy, has not been reported. In a prior study, we used a tail spike protein from a bacteriophage (ΦAB6TSP) that digests Pse-containing exopolysaccharide (EPS) from Acinetobacter baumannii strain 54149 (Ab-54149) to form a glycoconjugate for preparing anti-Ab-54149 EPS serum. We report here that a catalytically inactive ΦAB6TSP (I-ΦAB6TSP) retains binding ability toward Pse. In addition, an I-ΦAB6TSP-DyLight-650 conjugate (Dy-I-ΦAB6TSP) was more sensitive in detecting Ab-54149 than an antibody purified from anti- Ab-54149 EPS serum. Dy-I-ΦAB6TSP also cross-reacted with other pathogenic bacteria containing Pse on their surface polysaccharides (e.g., Helicobacter pylori and Enterobacter cloacae), revealing it to be a promising probe for detecting Pse across bacterial species. We also developed a detection method that employs I-ΦAB6TSP immobilized on microtiter plate. These results suggested that the anti-Ab-54149 EPS serum would exhibit cross-reactivity to Pse on other organisms. When this was tested, this serum facilitated complement-mediated killing of H. pylori and E. cloacae, indicating its potential as a cross-species antibacterial agent. This work opens new avenues for diagnosis and treatment of multidrug resistant (MDR) bacterial infections.

Autophagy upregulation and apoptosis downregulation in DAHP and triptolide treated cerebral ischemia.

It has previously been demonstrated that ischemic stroke activates autophagy pathways; however, the mechanism remains unclear. The aim of this study is to further investigate the role that autophagy plays in cerebral ischemia. 2, 4-diamino-6-hydroxy-pyrimidine (DAHP), for its nitric oxide synthase (NOS) inhibiting neuroprotective effect, and triptolide (TP), for its anti-inflammatory property, were selected to administer pre middle cerebral artery occlusion (MCAO). The drugs were administered 12 hours prior to MCAO. Both magnetic resonance imaging (MRI) and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining showed that the drugs reduce the area of infarction. Immunoblotting analysis revealed increases in Beclin-1 and myeloid cell leukelia-1(Mcl-1) in treated rats. This could be a contributing factor to the reduction in autophagy induced damage. Immunochemistry and western blot showed that mTOR expression in treated rats was marginally different 24 h after injury, and this could also be significant in the mechanism. Furthermore, terminal deoxynucleotidyl transferase- (TdT-) mediated dUTP nick end labeling (TUNEL) staining proved that the drugs are effective in reducing apoptosis. The upregulation of Beclin-1 and Mcl-1 and downregulation of Bcl-2, caspase-3, and the Bcl-2/Beclin-1 ratio infer that the neuroprotective effect of DAHP and TP act via the mediation of autophagy and apoptosis pathways.

Inhibition of GTP cyclohydrolase reduces cancer pain in mice and enhances analgesic effects of morphine.

Noncoding polymorphisms of the GTP cyclohydrolase gene (GCH1) reduce the risk for chronic pain in humans suggesting GCH1 inhibitors as analgesics. We assessed the effects of the GCH1 inhibitor diaminohydroxypyrimidine (DAHP) on nociception and inflammation in a mouse melanoma and a sarcoma cancer pain model, and its co-effects with morphine in terms of analgesic efficacy and respiratory depression. GCH1 inhibition did not reduce the tumor-evoked nociceptive hypersensitivity of the tumor-bearing paw. However, DAHP reduced melanoma- and sarcoma-evoked systemic hyperalgesia as determined by analyzing contralateral paws. GCH1 inhibition increased the inflammatory edema and infiltration with polymorphonuclear leukocytes surrounding the tumor but reduced the tumor-evoked microglia activation in the spinal cord suggesting that an increase of the local immune attack against the tumor may avoid general pain hypersensitivity. When used in combination with morphine at high or low doses, GCH1 inhibition increased and prolonged the analgesic effects of the opioid. It did not, however, increase the respiratory depression caused by morphine. Conversely, the GCH1-product, tetrahydrobiopterin, caused hyperalgesia, antagonized antinociceptive effects of morphine, and aggravated morphine-evoked respiratory depression, the latter mimicked by a cGMP analog suggesting that respiratory effects were partly mediated through the BH4-NO-cGMP pathway. The observed effects of GCH1 inhibition in the tumor model and its enhancement of morphine-evoked antinociception without increase of morphine toxicity suggest that GCH1 inhibitors might be useful as co-therapeutics for opioids in cancer patients.

[Comparison of the efficacy of two different iron supplements for anemia prevention in piglets]. / Vergelijkend onderzoek naar de werkzaamheid van twee verschillende ijzersupplementen voor anemiepreventie in biggen.

In a randomized, confirmatory study performed between July and October 2000 the efficacy of two iron products in preventing iron deficiency anaemia was compared. A total of 102 newborn piglets from ten litters were treated intramuscularly with 200 mg iron as iron dextran per ml, or 200 mg iron as gleptoferron per ml. For true comparison, piglets within a litter of a sow were subdivided into pairs on the basis of birth weight (one pair of the two heaviest piglets, et cetera). Within a pair, treatment with the iron supplements was randomly allocated. One group of piglets was injected at an age of 1 day (experiment 1) and the other group of piglets was injected at an age of 3 days (experiment 2). The piglets were weighed and blood samples were taken at an age of 18 days (experiment 1) or at an age of 19 days (experiment 2). Average daily weight gain and haemoglobin concentrations of both treatment groups were compared. Both products were very effective in preventing anaemia. No significant differences could be found between the two formulations. It can be concluded that iron-dextran and gleptoferron can be used with similar effect for anaemia prevention in piglets.

Synergistic interaction between 13-cis-retinoic acid and glucarate: activity against rat mammary tumor induction and MCF-7 cells.

At high dietary levels in vivo, both 13-cis-retinoic acid and calcium glucarate inhibit the induction of rat mammary tumors by 7,12-dimethylbenz(a)anthracene. The present study shows that sub-optimal dietary levels of each, which individually have no effect on tumor induction, when combined together in the diet, significantly increases tumor latency and suppresses tumor frequency in the rat system. Weight gain of animals was similar in control and experimental groups. Furthermore, ineffective sub-optimal dosages of glucarate and 13-cis-retinoic acid interacted synergistically to inhibit the growth in vitro of the MCF-7 human breast cancer cells. By varying the concentrations of glucarate and 13-cis-retinoic acid independently, evidence was obtained that in combination glucarate may play an adjuvant role, with the retinoid as the effector. Thus, the results of this experimental animal study demonstrate for the first time the potential use in synergistic combination of 2 normal metabolites in non-toxic chemoprevention and chemotherapy.

Copper heptonate for the treatment of hypocupraemia in sheep.

Trials were carried out to compare the efficacy of copper heptonate with other available therapeutic products. On nine farms with a copper deficiency problem, over 2000 ewes were treated in mid-pregnancy. All the treatments increased whole blood copper levels and plasma caeruloplasmin (ferroxidase) activity in ewes, and protected their lambs from swayback. The lambs from ewes treated with copper heptonate and copper oxide needles had significantly higher blood copper levels than lambs from untreated control ewes. Treatment with copper heptonate was shown to have a therapeutic index of at least 5 in Swaledale ewes of normal copper status. In two groups of Welsh mountain sheep from different backgrounds, both with normal blood copper levels before treatment, the therapeutic indices were 3 to 5.

Improvement of disability and akinesia of patients with Parkinson's disease by intravenous iron substitution.

Akinetic crises are one of the problems arising in patients with Parkinson's disease in particular after long term treatment with levo-dihydroxyphenylalanine (L-DOPA). They are characterized by severe disability to move. Increasing dosages of L-DOPA and decarboxylase or monoaminooxidase inhibitors do not improve these symptoms. Intravenously applied iron in the form of a ferri-ferro-complex exhibits a considerable benefit for all patients treated so far. They regained a remarkable mobility. Their disability score dropped from up to 90 percent down to 30 percent. The effect is dosage-dependent, and withdrawal of iron will lead again to akinetic crises.

Serum ferritin and total iron-binding capacity to estimate iron storage in pigs.

The inability to accurately determine storage iron in baby pigs limits the development of new treatment programs. In pigs treated neonatally with iron dextran, serum ferritin had increased dramatically at ten days of age and then returned to near preinjection levels by 50 days of age. In contrast, serum ferritin in untreated pigs declined until they were offered creep feed at 21 days of age. When serum ferritin, serum iron, serum total iron-binding capacity, erythrocyte number, packed cell volume, and blood hemoglobin were measured in three-week-old pigs, serum ferritin combined with serum total iron-binding capacity correlated significantly with the total nonheme iron in the liver and spleen. The nonheme iron (in mg) could be predicted (r2 = 0.71) by the following expression: 8.7 + 0.6 (ferritin in ng/ml).

[Clinical study on prophylaxis of diacetyl-glucaro-(1-4) (6-3) dilactone for recurrence of bladder cancer].

In order to investigate the effect of Diacetyl-glucaro-(1-4) (6-3) dilactone to prevent post-operative recurrence of bladder cancer, we estimated the recurrent rate of 34 patients who were diagnosed as having bladder cancer and treated by several surgical methods and oral administration of the drug in our hospital during 1971 and 1980. The following results were obtained: The 6, 12, 18, 24, 30, 36 and over 36 months-recurrent rates were 3.1%, 26.4%, 32.5%, 32.5%, 32.5%, 46% and 46%. In control group, they were 10%, 23.3%, 34.8%, 46.2%, 73.1%, 73.1% and 100% In a 24 month follow-up, there was no difference of recurrent rate between the group receiving the drug and control group: a recurrent rate of administrated group was lower than that of the control after 24 months. The recurrent rate of the group receiving Diacetyl-glucaro-(1-4) (6-3) dilactone combined with instillation was lower in a 2 year follow-up. It was anticipated that the combined therapy (Diacetyl-glucaro-(1-4) (6-3) dilactone administration and intravesical instillation of anticancer drugs) was useful in order to prevent recurrence of bladder cancer.

Animal studies on the reduction of aminoglycoside-induced nephrotoxicity by D-glucaro-1,5-lactam.

We studied the effect of D-glucaro-1,5-lactam on aminoglycoside-induced nephrotoxicity in rats. Parameters of nephrotoxicity were urinary excretion of tubule cells and malate dehydrogenase. When given in appropriate doses, either i. m. or via an oral tube, D-glucaro-1,5-lactam significantly reduced the excretion of cells and enzymes during the administration of gentamicin, tobramycin, dibekacin, netilmicin and ribostamycin. It did not impair the therapeutic efficacy of ribostamycin in the experimental treatment of acute pyelonephritis in rats. The protective effect of D-glucaro-1,5-lactam could be ascribed to its inhibition of beta-glucuronidase, an enzyme which is located in renal lysosomes and which is activated by aminoglycosides.

Comparison of gleptoferron with iron dextran for anemia prevention in young pigs.

Gleptoferron, a sterile aqueous colloidal solution of beta-ferric oxyhydroxide and dextran glucoheptonic acid, was compared with iron dextran for the prevention of Fe deficiency anemia in young pigs. Using 26 litters, pigs (within each litter) were randomly allotted to one of three treatments: 1) control (no Fe), 2) iron dextran (200 mg) and 3) gleptoferron (200 mg). Blood was collected at 0, 10, 21 and 50 d post-treatment for red blood cell count (RBC), hematocrit (HCT), hemoglobin (HGB) concentration, serum Fe concentration (Fe) and serum Fe-binding capacity (IBC). At 21 d, 30 pigs (one pig/treatment from each of 10 litters) were killed to determine milligrams nonheme Fe (NHFe) in liver and spleen, bile IBC and concentrations of bile and fecal Fe. There were no differences (P greater than .05) between Fe sources in 3- or 8-wk body weight or in any of the blood or tissue characteristics. In contrast, control pigs gained less (P less than .05) weight and had lower (P less than .05) RBC, HGB, HCT, serum Fe and liver and spleen NHFe than those that received iron dextran or gleptoferron. Serum IBC was greater (P less than .05) for the control than for Fe-treated pigs. These results demonstrate that the iron from iron dextran and gleptoferron is used with similar efficiency for anemia prevention in young pigs.

Parathyroidectomy in chronic renal failure.

Parathyroidectomy was carried out in 26 patients over a 14-year period. Excellent results were obtained in patients with severe hyperparathyroidism. Vascular calcification, hypercalcaemia and pruritus did not justify surgery unless associated with unequivocal hyperparathyroidism. 13 patients required intravenous calcium infusion for up to 2 weeks to control post-operative hypocalcaemia. Calcium requirements could be predicted from the pre-operative plasma alkaline phosphatase level. Following operation continued treatment with vitamin D was necessary to prevent hypocalcaemia. Hyperparathyroidism recurred in 1 patient after 8 years and 4 patients developed osteomalacia. Since parathyroid hormone may have toxic effects other than those on bone, maintenance of normal levels should be a long-term objective in the treatment of patients with chronic renal failure. Where large parathyroid glands are present, surgical reduction in gland mass is a logical prelude to long-term suppression of parathyroid hormone with vitamin D and phosphate-binding agents.

Comparative effects of calcium chloride and calcium gluceptate.

Calcium chloride and calcium gluceptate were compared in their ability to increase plasma ionized calcium concentrations ([Ca2+]). To correct a low ionized calcium concentration, each of 10 critically ml of a 10% solution, containing elemental calcium 27 mg ml-1) and calcium gluceptate (20 ml, containing elemental calcium 18 mg ml-1) over a 5-min period in randomized order approximately 6 h apart. [Ca2+] and haemodynamic variables (mean arterial pressure (MAP), mean right atrial pressure (RAP) and heart rate (HR)) were monitored for a 30-min period following completion of calcium infusion. Infusion of either calcium preparation was associated with similar increases in [Ca2+] (5 min after infusion of calcium chloride: 33 +/- 3.1%; calcium gluceptate: 32 +/- 4.3% (mean +/- SEM)) and the effects on MAP were similar for each solution (11.1 +/- 1.8% and 9.7 +/- 2.4%, respectively).

Randomized double-masked trial of sodium oxyferriscorbone for the treatment of gastric ulcers.

A double-blind 4-week trial of sodium oxyferriscorbone versus placebo (distilled water) was conducted in 46 outpatients with endoscopically confirmed gastric ulcer. Ulcer healing occurred in 15 of 20 patients receiving sodium oxyferriscorbone (75%) and in 7 of 20 patients receiving placebo (35%). Patients receiving sodium oxyferriscorbone experienced less pain and required less antacid than those receiving placebo (P less than 0.05). Side effects were reported in 12 patients, 7 while receiving sodium oxyferriscorbone and 5 while receiving placebo. Six patients did not complete the study due to ulcer complications. Routine laboratory tests revealed no persistent abnormalities that could be related to the treatment. Five placebo-treated patients that were therapeutic failures were switched to sodium oxyferriscorbone and healing was observed within 3 weeks. It is concluded that sodium oxyferriscorbone is effective in enhancing healing of gastric ulcers.

Protective effect of D-glucaro-delta-lactam against amino-glycoside-induced nephrotoxicity in rats.

The nephrotoxicity of rats caused by dibekacin (3',4'-dideoxykanamycin B) or kanamycin with or without dextran was effectively reduced by D-glucaro-delta-lactam potassium salt, as evidenced by lower levels of blood urea nitrogen and kidney edema rate, better excretion of antibiotics,and less morphological damage. Protection was dosage related, and potentiated with increasing doses, but only when the two drugs were given simultaneously. Among three alkali-metal salts examined, the potassium salt was almost equal to the lithium salt, but surpassed the sodium salt in effectiveness. Inorganic salts, in particular potassium chloride were found to be effective for the protection of normal rats, but their effect decreased for the dehydrated rats, especially in the presence of dextran.

Effect of D-Glucarates on basic antibiotic-induced renal damage in rats.

Dehydrated rats regularly develop acute renal failure following single injection of aminoglycoside antibiotics combined with dextran or of antibiotics only. Oral administration of 2,5-di-O-acetyl-D-glucaro-1,4-6,3-dilactone protected rats against renal failure induced by kanamycin-dextran. The protective effect was prevalent among D-glucarates, and also to other saccharic acid, hexauronic acids and hexaaldonic acids, although to a lesser degree, but not to a hexaaldose, sugar alcohols, substances inthe TCA cycle and other acidic compounds. D-Glucarates were effective against renal damage induced by peptide antibiotics as well as various aminoglycoside antibitocis. Dose-responses were observed in the protective effect of D-Glucarates. With a D-glucarate of a fixed size of dose, approximately the same degree of protection was obtained against renal damages induced by different basic antibiotics despite large disparities in administration doses of different antibiotics. D-Glucarates had the ability to prevent renal damage but not to cure it. Rats excreted acidic urine when they were spared from renal lesions by monosaccharides. The reduction effect of D-glucarates against nephrotoxicity of basic antibiotics was discussed.