Therapeutic potential of phages in autoimmune liver diseases.

Autoimmune liver disease (ALD) poses a difficult medical challenge, as there is a significant number of patients in whom current therapy offers questionable or no benefit, yet its side effects may be serious, including the development of malignancy. Bacterial viruses (phages) have been recognized increasingly as immunomodulators contributing to immune homeostasis and curbing inflammation. Accumulating data suggest that phages may be useful in immunotherapy of ALD. Phages have been shown to down-regulate the expression and/or production and activity of factors associated with hepatic injury [reactive oxygen species, Toll-like receptor (TLR)-4 activation, nuclear factor kappa B (NF-κB) activation, proinflammatory and procoagulant activities of platelets] and up-regulate the expression and/or production of factors demonstrated as playing a protective role [interleukin (IL)-10, IL-1 receptor antagonist].

Phages in the global fruit and vegetable industry.

From recent articles, we have learned that phages can constitute a promising alternative in the food industry to eliminate bacterial pathogens from seedlings in greenhouse and field environments, as well as from fresh-cut food products. The fruit and vegetable industry requires quite a different approach than the meat or dairy industry. Several factors can inhibit efficacy of phage treatment such as plant watering or washing ready-to-eat products (water may dilute therapeutic doses), UV irradiation or extensive spreading of phytopathogens by wind, insects or even humans. Spontaneously occurring anomalous weather conditions in different parts of the world also may have an enormous impact on phage persistence in cultivations and on yields. Despite that, some phage preparations are commercially available and, without doubt, are much safer than chemical treatments. Along with increasing worldwide fruit and vegetable consumption, plant diseases and human foodborne illnesses are becoming a serious economic problem, resulting in a focus on optimization of phage treatment.

Application of microbiological quantitative methods for evaluation of changes in the amount of bacteria in patients with wounds and purulent fistulas subjected to phage therapy and for assessment of phage preparation effectiveness (in vitro studies).

PURPOSE: Quantitative microbiological studies may provide important information required for successful phage therapy (PT), however methods for PT monitoring of purulent wounds and fistulas has never been reported before. Therefore our goal was to determine and apply microbiological quantitative methods (MQMs) for monitoring experimental PT. METHODS: Samples from agar plates with growing bacteria were collected using dry and wet sterile compresses, or swabs. After shaking the sample in saline the amount of bacteria in suspension was determined. The method was standardized. The MQM using compress was applied for comparison of in vitro activity of phage preparations with other agents for wound rinsing. The usefulness of this swabbing method was tested in the Phage Therapy Unit for monitoring of experimental PT of patients with chronic wounds or purulent fistulas. RESULTS: Minimum, maximum and standard deviation values used for standardization of the studied method showed that data repeatability was good; thus the method was used for quantitation of bacteria taken both from plates in vitro and patients samples. Effectiveness of phage preparations was compared to gentamicin in vitro. Phages were as effective as antibiotics in reducing the amount of bacteria on agar plates, and this effect was not only due to simple mechanical removal of bacteria, but dependent on their antibacterial activity. We have also observed that the results of bacteria quantitation may correlate with the local status of a wound/fistula in a particular stage of PT. CONCLUSION: The standardized swabbing method of bacteria quantitation can be used for PT monitoring. Presented MQMs are simple and may help to monitor the therapy process and to decide on its duration, frequency and a kind of the phage applied. They can also be applied in other antibacterial treatment strategies.

Eradication of Enterococcus faecalis by phage therapy in chronic bacterial prostatitis--case report.

The treatment of three patients suffering from chronic bacterial prostatitis who were qualified for an experimental phage therapy protocol managed at the Phage Therapy Unit in Wroclaw is described. They had previously been treated unsuccessfully with long-term targeted antibiotics, autovaccines, and laser biostimulation. Rectal application of phage lysates targeted against Enterococcus faecalis cultured from the prostatic fluid gave encouraging results regarding bacterial eradication, abatement of clinical symptoms of prostatitis, and lack of early disease recurrence.

Successful eradication of methicillin-resistant Staphylococcus aureus (MRSA) intestinal carrier status in a healthcare worker--case report.

We describe bacteriophage therapy in the case of a healthcare worker whose gastrointestinal tract was colonized by methicillin-resistant Staphylococcus aureus (MRSA) with subsequent urinary tract infection caused by the same pathogen. Oral treatment with anti-MRSA phages resulted in eradication of the carrier status.

Bacteriophages and transplantation tolerance.

Our recent findings suggest that bacteriophages (phages) may not only eliminate bacteria, but also modulate immune functions. In this communication, we demonstrate that phages may strongly inhibit human T-cell activation and proliferation as well as activation of the nuclear transcription factor NF-kappaB in response to a viral pathogen. Phage administration in vivo can diminish cellular infiltration of allogeneic skin allografts. Thus, phage treatment should be considered in antibiotic-resistant posttransplantation infections. Furthermore, phages could find a broader application in clinical transplantation.

Alternative therapies in antibiotic-resistant infection.

CASE REPORT: A 24-year-old woman suffering from post-influenza otitis media infection was initially treated with several series of a steroid (Elocon) and a combination of steroids and antibiotics (Atecortin, Dicortineff) without significant medical benefit. The isolated bacterial strains were identified as Staphylococcus homis and Staphylococcus epidermidis. Specific phage therapy applied sequentially over a period of three weeks resulted only in a partial reduction in inflammation and limited improvement in overall health condition. Oral application of lactoferrin (LF; 50-mg daily oral doses for seven days with two-week intervals) led to a complete clearance of both bacterial strains and full recovery of the patient. The recovery was associated with increased myelopoiesis and a sustained elevation of serum endogenous LF. In conclusion, specific bacteriophage therapy combined with the administration of lactoferrin proved to be effective in the treatment of antibiotic-resistant external ear infection.

Antitumor activity of bacteriophages in murine experimental cancer models caused possibly by inhibition of beta3 integrin signaling pathway.

Bacteriophages (phages) as bacterial viruses are generally believed to have no intrinsic tropism for mammalian cells. In this study the interactions between phages and various eukaryotic cells were investigated. Binding of phages to the membranes of cancer and normal blood cells was observed. Moreover, it was shown that the wild-type phage T4 (wtT4) and its substrain HAP1 with enhanced affinity for melanoma cells inhibit markedly and significantly experimental lung metastasis of murine B16 melanoma cells by 47% and 80%, respectively. A possible molecular mechanism of these effects, namely a specific interaction between the Lys-Gly-Asp motif of the phage protein 24 and beta3-integrin receptors on target cells is proposed. It was also shown that anti-beta3 antibodies and synthetic peptides mimicking natural beta3 ligands inhibit the phage binding to cancer cells. This is in line with the well-described beta3 integrin-dependent mechanism of tumor metastasis. It is concluded that the blocking of beta3 integrins by phage preparations results in a significant decrease in tumor invasiveness.

New insights into the possible role of bacteriophages in transplantation.

Due to the increasing prevalence of drug-resistant bacterial infections in the "post-antibiotic era," bacteriophages (bacterial viruses, BP) may be useful to administer to transplant recipients without exposing them to an increased risk of rejection, which occurs consequent to some viral infections. Herein we present evidence that at least some coliphages (T4) do not pose such risk. Interestingly, they may produce immunosuppressive effects extending transplant survival. Our data suggest that BP may be used in clinical transplantation to treat drug-resistant bacterial infections and perhaps as an adjunct to immunosuppressive therapy.

[Colonization of post-operative wounds by endogenous gram-negative rods in patients after laryngectomy]. / Kolonizacja ran pooperacyjnych endogennymi paleczkami gram-ujemnymi u pacjentów po chirurgicznym usunieciu raka krtani.

The bacterial flora of the posterior laryngeal wall, larynx and surgical wounds of the patients treated surgically for carcinoma of the larynx was investigated. A similarity of some bacterial strains isolated from various habitats was demonstrated by comparing the biochemical characteristics, drug resistance patterns, and susceptibility to bacteriocins and bacteriophages. Bacteriophagotyping confirmed the similarity of rods belonging to the genera Enterobacter and Pseudomonas. Thus, colonization of surgical wounds by bacteria present in the pharynx and the larynx, but not inducing healing complications, has been confirmed. The detected strains demonstrated high susceptibility to antibiotics, including those administered within the framework of post-operative preventive treatment.

Effective phage therapy is associated with normalization of cytokine production by blood cell cultures.

The aim of this study was to investigate the effect of phagotherapy on tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) serum levels and the ability of blood cells to produce these cytokines in culture. Fifty one patients with long-term, suppurative infections of various tissues and organs were enrolled. The ability of cells to secrete cytokines was tested using whole blood cell cultures, unstimulated or stimulated with lipopolysaccharide (LPS) from E. coli. In addition, cytokine serum levels were determined. Measurement of cytokine activity was performed using bioassays. We showed that TNF-alpha, but not IL-6 serum levels, were regulated upon division of patients into categories exhibiting initial: low, moderate and high cytokine levels. The low spontaneous production of IL-6 by blood cell cultures was elevated significantly on day 21 of phage therapy, whereas high release of this cytokine was inhibited. No such correlation was observed with LPS-induced IL-6 production in cell cultures when cells from low-, moderately- or highly-reactive patients were studied. Phage therapy modified TNF release according to the initial ability to produce that cytokine: it reduced TNF production in high responders and increased it in low responders. Patients infected only with Gram-positive bacteria demonstrated analogous changes in the spontaneous and LPS-induced TNF-alpha production as in the whole studied group. A similar kind of regulation was observed in TNF-alpha and LPS-induced production, i.e. low production was significantly elevated, high strongly inhibited, and moderate only slightly affected. In summary, we demonstrated for the first time that effective phage therapy can normalize TNF-alpha serum levels and the production of TNF-alpha and IL-6 by blood cell cultures.

Bacteriophage therapy of bacterial infections: an update of our institute's experience.

1307 patients with suppurative bacterial infections caused by multidrug-resistant bacteria of different species were treated with specific bacteriophages (BP). BP therapy was highly effective; full recovery was noted in 1123 cases (85.9%). In 134 cases (10.9%) transient improvement was observed and only in 50 cases (3.8%) was BP treatment found to be ineffective. The results confirm the high effectiveness of BP therapy in combating bacterial infections which do not respond to treatment with the available antibiotics.

[Successful treatment with bacteriophage in purulent cerebrospinal meningitis in a newborn]. / Skuteczne zastosowanie bakteriofagoterapii w ropnym zapaleniu opon mózgowo-rdzeniowych u noworodka.

The subject of this report is the case of purulent meningitis in new-born caused by Klebsiella pneumoniae. As the intensive antibiotic therapy turned out to be ineffective phage therapy was applied. Oral administration of specific phage preparate for the period of 5 weeks resulted in complete sterilization of cerebrospinal fluid and unquestionable improvement of child's health. However, after several ventriculopunctures some complications appeared (haemorrhage into central nervous system, extra infection). They were treated in standard way. Because of increasing internal hydrocephalus and necessity of operation, the child was sent to suitable hospital for further treatment.

[The study on biology of bacteriophages and their usage in the treatment of bacterial diseases and on the influence of different bacteriophages on cytokine production by leukocytes in human peripheral blood]. / Badania nad biologia bakteriofagów i ich wykorzystaniem w leczeniu zakazen bakteryjnych oraz wplywu róznych bakteriofagów na wytwarzanie cytokin przez leukocyty krwi obwodowej czlowieka.

The authors showed the examinations of the biology bacteriophages and using them in the treatment of the bacteriology infection and influence difference bacteriophages in producing cytokinins by leukocytes of human peripheral blood.

Acute infarction of the left ventricular papillary muscle: electrocardiographic pattern and recognition of its location.

BACKGROUND AND HYPOTHESIS: ST-segment depression during acute myocardial infarction (AMI) is known to herald serious hemodynamic complications. Since the mechanism of this dependence is not clear, we reinvestigated the old concept of papillary muscle infarction (PMI) as a cause of marked ST depression. METHODS: Autopsies and morpho-electrocardiographic cor-relations were performed in 53 patients with AMI involving one or both left ventricular papillary muscles, and in 10 patients with AMI, but without acute PMI. RESULTS: ST-segment depression > or = 1 mm in at least two leads (mean 3.6 +/- 2.2 mm) was found in 46 (86.8%) patients with, and in one without acute PMI. Thus, the sensitivity and specificity in selecting patients with acute PMI from among those with AMI were 86.8 and 90%, respectively, with an overall accuracy of diagnosis of acute PMI in the course of AMI of 87.3%. Among 26 patients with ST elevation consistent with diagnosis of AMI, ST depression, recorded in 22 patients, was insignificantly greater than in 24 of 27 patients without ST elevation: 4.1 +/- 2.9 versus 3.1 +/- 1.2 mm. Localization of ST depression in the limb leads allowed recognition of which papillary muscle suffered from acute infarction: ST depression in the inferior leads was seen only in patients with anterolateral PMI, whereas in leads I and/or a VL it was seen only in cases with posteromedial PMI. This rule was also valid in patients without concomitant ST elevation. CONCLUSION: Patients with acute PMI show marked ST-segment depression. Its location in the limb leads allows recognition of which papillary muscle has undergone necrosis. This cause of marked ST depression in patients with AMI may explain the high mortality in this particular group.

Parasympathetic withdrawal precedes spontaneous blood pressure elevations in women with primary hypertension.

Exaggerated sympathetic activity is widely accepted as one of the fundamental mechanisms leading to primary hypertension and being responsible for frequent episodes of blood pressure elevation in hypertensive patients. Some data suggest also that basal parasympathetic tone in this entity is lowered. However, the effects of autonomic nervous activity on heart rate variability during spontaneous blood pressure elevations have not been yet evaluated. That is why we present the preliminary results of 24-hour electrocardiogram and blood pressure monitoring in 13 women with mild primary hypertension and with 25 episodes of blood pressure elevations. Time- and frequency domain measurements of heart rate variability found during the 24- and 10- hour daily periods were compared with those obtained during four 5-min records: 25-20, 15-10 min and immediately before, as well as immediately after the recording of blood pressure elevation. Significant decrease in parameters representing vagal tone was found during 5-min periods not only immediately preceding or following blood pressure elevations, but also 10 and 20 min before these episodes. Moreover, low-frequency component of heart rate variability was significantly lowered 10 min before and immediately after the recording of blood pressure elevation. These results suggest that among various pathogenetic mechanisms of spontaneous blood pressure elevations in women with primary hypertension, sudden withdrawal of parasympathetic tone should be taken into account.

Heart rate variability before sudden blood pressure elevations or complex cardiac arrhythmias in phaeochromocytoma.

In phaeochromocytoma, sudden hypertensive or arrhythmic episodes are believed to be associated with excessive free catecholamine excretion. However, lack of correlation between blood pressure (BP) and plasma catecholamine levels has been reported. Therefore an attempt was made to assess the sympathovagal balance before and during episodes of BP elevation or complex cardiac arrhythmias in this disease. Ten patients with phaeochromocytoma and 10 matched controls with essential hypertension underwent simultaneous 24 h Holter ECG and BP monitoring. BP elevation was diagnosed when the BP exceeded the mean 24 h values by 40 mm Hg systolic or 30 mm Hg diastolic, respectively. Heart rate variability (HRV) was measured for 5 min periods 1 h before, 15 min before and during 13 episodes of BP elevation in phaeochromocytoma and 13 episodes in the control group, as well as at 1 h, 15 min and immediately before five arrhythmic events in phaeochromocytoma. In phaeochromocytoma, vagal activity measured 1 h before BP elevation was markedly higher than in control hypertensives. However, in both groups at 15 min before and during the hypertensive events, the vagal tone decreased significantly. In contrast, just before the arrhythmic events HRV remained unaltered with a slight insignificant increase in sympathetic activity. We conclude that in phaeochromocytoma, pronounced BP elevations during daily activities are preceded by a parasympathetic withdrawal, similar to the findings in essential hypertension. Such a sequence does not seem to precede sudden complex arrhythmic events in phaeochromocytoma.