Absence of Toxemia in Clostridioides difficile Infection: Results from Ultrasensitive Toxin Assay of Serum.

Clostridioides difficile infection (CDI) is caused by Toxins A and B, secreted from pathogenic strains of C. difficle. This infection can vary greatly in symptom severity and in clinical presentation. Current assays used to diagnose CDI may lack the required sensitivity to detect the exotoxins circulating in blood. The ultrasensitive single molecule array (Simoa) assay was modified to separately detect toxin A and toxin B in serum with a limit of detection at the low picogram level. When applied to a diverse cohort, Simoa was unable to detect toxins A or B in serum from patients with CDI, including many classified as having severe disease. The detection of toxin may be limited by the inference of antitoxin antibodies circulating in serum. This result does not support the hypothesis that toxemia occurs in C. difficile infection, conflicting with the findings of other published reports.

Toxemia in Human Naturally Acquired Botulism.

Human botulism is a severe disease characterized by flaccid paralysis and inhibition of certain gland secretions, notably salivary secretions, caused by inhibition of neurotransmitter release. Naturally acquired botulism occurs in three main forms: food-borne botulism by ingestion of preformed botulinum neurotoxin (BoNT) in food, botulism by intestinal colonization (infant botulism and intestinal toxemia botulism in infants above one year and adults), and wound botulism. A rapid laboratory confirmation of botulism is required for the appropriate management of patients. Detection of BoNT in the patient's sera is the most direct way to address the diagnosis of botulism. Based on previous published reports, botulinum toxemia was identified in about 70% of food-borne and wound botulism cases, and only in about 28% of infant botulism cases, in which the diagnosis is mainly confirmed from stool sample investigation. The presence of BoNT in serum depends on the BoNT amount ingested with contaminated food or produced locally in the intestine or wound, and the timeframe between serum sampling and disease onset. BoNT levels in patient's sera are most frequently low, requiring a highly sensitive method of detection. Mouse bioassay is still the most used method of botulism identification from serum samples. However, in vitro methods based on BoNT endopeptidase activity with detection by mass spectrometry or immunoassay have been developed and depending on BoNT type, are more sensitive than the mouse bioassay. These new assays show high specificity for individual BoNT types and allow more accurate differentiation between positive toxin sera from botulism and autoimmune neuropathy patients.

Hyponatraemia compounding pre-eclamptic toxaemia in a patient with type 1 diabetes.

We report a case of profound, symptomatic hyponatraemia in association with pre-eclamptic toxaemia (PET) in a 38-year-old nulliparous woman with type 1 diabetes mellitus. This patient developed hypertension and proteinuria at 31+6 weeks' gestation and was admitted for management of pre-eclampsia. Severe headache, visual disturbance and nausea were associated with a hyponatraemia of 115 mmol/L followed by ketoacidosis. This was reversed through fluid restriction, supplementation with 1.8%-3.0% hypertonic saline and a volume-reduced variable-rate insulin infusion. Clinical stability was achieved and she was subsequently worked up for an induction of labour for worsening pre-eclampsia. Hyponatraemia in the context of PET has been previously reported as rare. However, it has complications that may significantly compound the sequelae of severe PET. We propose that specific and focused monitoring of serum sodium levels should become common practice in the management of women with this condition to allow for timely, measured correction of abnormalities.

Zeptomole per milliliter detection and quantification of edema factor in plasma by LC-MS/MS yields insights into toxemia and the progression of inhalation anthrax.

Inhalation of Bacillus anthracis spores can cause a rapidly progressing fatal infection. B. anthracis secretes three protein toxins: lethal factor (LF), edema factor (EF), and protective antigen (PA). EF and LF may circulate as free or PA-bound forms. Both free EF (EF) and PA-bound-EF (ETx) have adenylyl cyclase activity converting ATP to cAMP. We developed an adenylyl cyclase activity-based method for detecting and quantifying total EF (EF+ETx) in plasma. The three-step method includes magnetic immunocapture with monoclonal antibodies, reaction with ATP generating cAMP, and quantification of cAMP by isotope-dilution HPLC-MS/MS. Total EF was quantified from 5PL regression of cAMP vs ETx concentration. The detection limit was 20 fg/mL (225 zeptomoles/mL for the 89 kDa protein). Relative standard deviations for controls with 0.3, 6.0, and 90 pg/mL were 11.7-16.6% with 91.2-99.5% accuracy. The method demonstrated 100% specificity in 238 human serum/plasma samples collected from unexposed healthy individuals, and 100% sensitivity in samples from 3 human and 5 rhesus macaques with inhalation anthrax. Analysis of EF in the rhesus macaques showed that it was detected earlier post-exposure than B. anthracis by culture and PCR. Similar to LF, the kinetics of EF over the course of infection were triphasic, with an initial rise (phase-1), decline (phase-2), and final rapid rise (phase-3). EF levels were ~ 2-4 orders of magnitude lower than LF during phase-1 and phase-2 and only ~ 6-fold lower at death/euthanasia. Analysis of EF improves early diagnosis and adds to our understanding of anthrax toxemia throughout infection. The LF/EF ratio may also indicate the stage of infection and need for advanced treatments.

Effects of acute lipopolysaccharide-induced toxemia model on some neglected blood parameters.

The presence of lipopolysaccharide (LPS) in blood induces an inflammatory response which leads to multiple organ dysfunction and numerous metabolic disorders. Uncontrolled, improper or late intervention may lead to tissue hypoxia, anaerobic glycolysis and a disturbance in the acid -base balance. The effects of LPS-induced toxemia on biological and immunological markers were well studied. However, parameters such as base excess, ions, and acid-base balance were not fully investigated. Therefore, the objective of this study was to examine these blood parameters collectively in LPS-induced inflammatory toxemia in rat's model. After induction of toxemia by injecting LPS at a rate of 5 mg/kg body weight intravenously, blood was collected from the tail vein of twenty rats and immediately analyzed. After 24 hours, the animals were sacrificed and the blood was collected from the caudal vena cava. The results revealed that the levels of pH, bicar- bonate, partial pressure of oxygen, oxygen saturation, Alveolar oxygen, hemoglobin, hematocrit, magnesium (Mg2+), and calcium (Ca2+) were significantly decreased. On the other side, the levels of Base excess blood, Base excess extracellular fluid, partial pressure of carbon dioxide, lactate, Ca2+/Mg2+, potassium, and chloride were significantly increased compared to those found pre toxemia induction. However, sodium level showed no significant change. In conclusion, Acute LPS-toxemia model disturbs acid-base balance, blood gases, and ions. These parameters can be used to monitor human and animal toxemic inflammatory response induced by bacterial LPS conditions to assist in the management of the diagnosed cases.

Identification of toxemia in patients with Clostridium difficile infection.

Toxemia can develop in Clostridium difficile-infected animals, and correlates with severe and fulminant disease outcomes. Circumstantial evidence suggests that toxemia may occur in patients with C. difficile infection (CDI), but positive diagnosis is extremely rare. We analyzed the potential for C. difficile toxemia in patients, determined its characteristics, and assessed challenges. C. difficile toxins in serum from patients were tested using an ultrasensitive cell-based assay and further confirmed by Rac1 glucosylation assay. The factors that hinder a diagnosis of toxemia were assessed, including investigation of toxin stability, the level of toxins-specific neutralizing antibodies in sera and its effect on diagnosis limits. CDI patients develop detectable toxemia in some cases (2.3%). Toxins were relatively stable in stored sera. Neutralizing anti-toxin antibodies were present during infection and positively correlated with the diagnosis limits. Thus, the masking effect of toxin-specific neutralizing antibodies is the major obstacle in diagnosing C. difficile toxemia using cell-based bioassays.

[The efficiency of reamberinum in infusion therapy of acetonemic syndrome in children].

The current paper presents the results of monitoring of 69 patients with acute respiratory viral infection, acetonemic syndrome. It was shown the effectiveness of inclusion of Reamberinum into complex therapy as a means for reduction of with acetonemic syndrome and for detoxic effect.

[Impact of the combined application of elastic bandaging of the lower extremities and autohemotransfusion on biochemical indices of the blood and severity of endogenous intoxication in patients with concomitant cardiovascular insufficiency].

Venous congestion in abdominal inner organs in surgical diseases in patients with heart insufficiency may additionally impact biochemical indices of the blood and severity of endogenous intoxication (EI). Basing on the data obtained in the investigations, th was established, that the lower extremities bandaging promotes exit of the blood from depot, where it resides in a concentrated state in cellular and biochemical aspects. It promotes more effective accomplishment of hemodilution as well as reduction of the EI severity in taking of the autologous blood.

Acid-base and electrolyte status during early induced pregnancy toxaemia in goats.

Eleven Murciano-Granadina goats in late pregnancy were separated into two groups (1) control (n=6) and (2) fasting for 72 h to induce pregnancy toxaemia (n=5). Venous blood was taken daily to determine acid-base and electrolyte parameters. Significant decreases in blood pH, bicarbonate concentration and base excess, and a significant increase in anion gap were observed after 24h of fasting. These changes were significantly correlated with non-esterified fatty acid concentration. No significant changes were observed in pCO(2), and electrolyte or lactate concentrations. Clinical signs of pregnancy toxaemia in fasted goats appeared by 72h post-fasting. These signs and the changes in acid-base balance disappeared once feed was reintroduced. Blood pH, bicarbonate concentration, base excess and anion gap could be indicators of early pregnancy toxaemia in goats.

Serologic evaluation of human microcystin exposure.

Microcystins are among the most commonly detected toxins associated with cyanobacteria blooms worldwide. Two episodes of intravenous microcystin exposures occurred among kidney dialysis patients during 1996 and 2001. Analysis of serum samples collected during these episodes suggests that microcystins are detectable as free and bound forms in human serum. Our goal was to characterize the biochemical evidence for human exposure to microcystins, to identify uncertainties associated with interpretation of these observed results, and to identify research needs. We analyzed serum samples using enzyme-linked immunosorbent assay (ELISA) methods to detect free microcystins, and gas chromatography/mass spectrometry (GC/MS) to detect 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB). MMPB is derived from both free and protein-bound microcystins by chemical oxidation, and it appears to represent total microcystins present in serum. We found evidence of free microcystins in patient serum for more than 50 days after the last documented exposure. Serum concentrations of free microcystins were consistently lower than MMPB quantification of total microcystins: free microcystins as measured by ELISA were only 8-51% of total microcystin concentrations as detected by the GC/MS method. After intravenous exposure episodes, we found evidence of microcystins in human serum in free and protein-bound forms, though the nature of the protein-bound forms is uncertain. Free microcystins appear to be a small but variable subset of total microcystins present in human serum. Research is needed to elucidate the human toxicokinetics of microcystins, in part to determine how observed serum concentrations can be used to estimate previous microcystin exposure.

Effect of ubiquinone-10 and succinic acid on functional characteristics of erythrocytes in rats with epinephrine toxemia.

Prophylactic dietary intake of synthetic ubiquinone-10, succinic acid, or mixture of these substances prevented disturbances in aggregation and electrophoretic mobility of erythrocytes and inhibited lipid peroxidation in cells of rats with experimental epinephrine-induced toxemia.

Sepsis and pathophysiology of anthrax in a nonhuman primate model.

Studies that define natural responses to bacterial sepsis assumed new relevance after the lethal bioterrorist attacks with Bacillus anthracis (anthrax), a spore-forming, toxigenic gram-positive bacillus. Considerable effort has focused on identifying adjunctive therapeutics and vaccines to prevent future deaths, but translation of promising compounds into the clinical setting necessitates an animal model that recapitulates responses observed in humans. Here we describe a nonhuman primate (Papio c. cynocephalus) model of B. anthracis infection using infusion of toxigenic B. anthracis Sterne 34F2 bacteria (5 x 10(5) to 6.5 x 10(9) CFU/kg). Similar to that seen in human patients, we observed changes in vascular permeability, disseminated intravascular coagulation, and systemic inflammation. The lung was a primary target organ with serosanguinous pleural effusions, intra-alveolar edema, and hemorrhagic lesions. This animal model reveals that a fatal outcome is dominated by the host septic response, thereby providing important insights into approaches for treatment and prevention of anthrax in humans.

[Endogenous intoxication in patients with stomach cancer observed immediately after surgery].

Severe endogenous intoxication (El) can trigger immediately after surgery many complications right up to multiple organ failure. The evaluation of El according to a degree of toxemia is most objective up to now. The purpose of the case study was to follow up the dynamics of the main El laboratory indices and to evaluate the correlation between them in patients with stomach cancer during the early postoperative period. The El parameters were determined in 2 hours before surgery as well as on days 1, 3, 5, 7 and 14 after surgery. The below indices were determined in blood of patients: toxicity index (TI), count of mean weight molecules (MWM), leukocytic intoxication index (LII) and total count of leukocytes (TCL). The study showed that, when patients with stomach cancer are admitted for surgical treatment, they have pronounced El, which intensifies even more postoperatively. The investigated laboratory indices are independently significant in the evaluation of the El degree. A reliable but faint correlation was found between the parameters of MWM and IT, and MWM and LII. TCL does not correlate reliably with other studied El parameters except for LII. The full-fledged evaluation of the El nature must be used in choosing purposefully detoxication methods with the aim of promoting the quality of postoperative treatment of patients with stomach cancer.

[Criteria for assessment of endogenous intoxication in burn trauma]. / Kriterii otsenki éndogennoi intoksikatsii pri ozhogovoi travme.

Registered in patients with burn trauma was the presence of endogenous intoxication (EI), which was determined by a higher value of coefficient CLP/AOS, a higher contents of medium-molecular peptides (MMP), a lower total and effective concentration of albumin (TAC and EAC) and by a reserve of the binding albumin ability. Intoxication coefficients, i.e. IC MMP/EAC and IC C/EAC showing the deposition and binding of toxic ligands, were acknowledged as the most informative EI parameters.

[Degree of endogenous intoxication in children with acute pneumonia in the presence of hypotrophy]. / Stepen' éndogennoi intoksikatsii u detei s ostroi pnevmoniei na fone gipotrofii.

Children with acute pneumonia in the presence of hypotrophy derive no benefit from conventional therapy, with no change for the better being noted in the degree of endogenous intoxication. Endotoxemia has been shown to be alleviated by endovascular laser irradiation of blood included in the complex of therapeutic means of remediation used in acute pneumonia against the background of hypotrophy, which fact permits reducing the volume of medicamentous therapy. The above treatment option has also been found to exert a beneficial effect on the course of the illness.

[Endogenic intoxication markers: data from capillary electrophoresis of trichloroacetic acid supernatant]. / Markery éndogennoi intoksikatsii po dannym kapilliarnogo élektrophoreza TKhU-supernatanta.

Capillary electrophoresis is suggested for characterization of trichloroacetic acid (TCA) supernatant in various visceral diseases involving endogenous intoxication. The technique of electrophoresis is described. Examples of electrophoregrams are offered with extrapolation to common clinical laboratory values. The appearance of a positive "cationic peak" instead of a negative systemic peak observed in health is a specific electrophoretic sign of endogenous intoxication. The area of the peak (proportionate to intoxication) and its ratio to the group undivided peak observed in the distal part of electrophoregram (minutes 8-14, anionic constituents) are significant. The values of the latter peak characterize the defense characteristics of the organism and are in proportion to them. Capillary electrophoresis is a perspective diagnostic method.

[Diagnostic significance of evaluating erythrocyte membrane permeability as an intoxication syndrome criterion]. / Diagnosticheskaia tsennost' otsenki pronitsaemosti membran éritrotsitov v kachestve kriteriia intoksikatsionnogo sindroma.

Twenty-five patients with pyoinflammatory maxillofacial diseases treated in an inpatient setting in the Regional Dentistry Clinic of Krasnodar and 8 healthy volunteers were examined. After opening and draining of the purulent focus the patients were divided into 2 groups. Group 1 consisted of 21 patients treated by intravenous 0.04% sodium hypochlorite (SHC) and intravenous infusions of SHC-oxidized autoblood for 2-3 days. Group 2 (6 pts) was treated by intravenous infusions of SHC-oxidized autoblood for 6 days. The criteria of intoxication were erythrocyte resistance tests in hypotonic sodium chloride and at different concentrations of urea in isotonic medium. A new method for evaluating erythrogram deviations from reference values is proposed, which helps adequately assess the severity of intoxication.

[A method for determining plasminogen with a Russian chromogenic substrate and its diagnostic significance]. / Metod opredeleniia plazminogena s otechestvennym khromogennym substratom i ego diagnosticheskoe znachenie.

Measurement of plasminogen, the key component of fibrinolysis system, is one of the basic methods for estimation of fibrinolysis. Methods based on the use of chromogenic substrates are often used in diagnosis. Plasminogen measurements are important for laboratory diagnosis of thrombophilia caused by deficiency or abnormalities of this fiber, for detection and evaluation of the DIC syndrome, and for monitoring the treatment by fibrinolytic preparations (streptokinase, t-PA, urokinase, etc.). An original chromogenic substrate having no foreign analogs has been created at Institute of Genetics and Selection of Industrial Microorganisms and Research Center of Hematology (Moscow). Unlike previously described plasmin substrates, pNa has been obtained by microbiological methods with Russian commercial enzymes subtilisine 72 and megaterine. This paper presents the results of plasminogen measurements in patients with DIC with the use of the original chromogenic substrate. The results were compared with those of tests with Berihrom-Plasminogen diagnostic kit (Behringwerke AG).

Deliganded albumin as a liquid adsorbent in the treatment of burn toxemia.

A new generation of synthetic carbon adsorbents was used in production of deliganded human serum albumin preparation. Thermal effects of officinal and deliganded albumin interaction with specific chemical markers were analyzed by flow microcalorimetry. The results demonstrated a 2.5-fold increase of the complexing ability for the deliganded one. The detoxifying potentials of deliganded albumin were studied in comparison with officinal preparation in rats with burn toxemia after IIIB-IV degree thermal injury and in model experiments with blood serum of patients after severe thermal burn. The transfusion of a 5% officinal albumin solution in rats 1 h after burn trauma resulted in a decrease of serum and liver cytosols cytotoxicity 2.2 and 2.4 times, respectively, in comparison with those of burned rats. After deliganded albumin transfusion the cytotoxic activity of blood serum dropped 8.5 times and that of the liver cytosols 18.5 times. The incubation of blood serum of injured patients with equal amounts of a 5% solutions of officinal or deliganded albumin resulted in a fall of the cytotoxicity level and the growth of binding ability. A comparative analysis of detoxifying potentials of albumin preparations has unambiguously demonstrated deliganded albumin advantages.

[New methods for assessing the severity of endogenous intoxication in surgical patients]. / Novye metody otsenki tiazhesti éndogennoi intoksikatsii u khirurgicheskikh bol'nykh.

New methods (a reticulocytic test, inhibition of erythrocyte sedimentation, registration of the frog's heart rate) were used in studying the toxicity of the serum, blood plasma and urine of surgical patients. The most informative methods for the assessment of the toxemia degree are thought to be the reticulocytic test and registration of the frog's heart rate. The maximum time for the determination of endotoxicity is 15 minutes. The pathological process severity can be estimated not only by the toxemia degree but also by its dynamics. The determination of endotoxicity is of value for choosing the methods for the individual detoxication therapy.