Pre-emptive analgesic effect of lornoxicam in mandibular third molar surgery: a prospective, randomized, double-blind clinical trial.

The aim of this study was to establish whether the pre-emptive use of lornoxicam (16mg) in third molar surgery ensures successful postoperative analgesia and reduces rescue analgesic intake when compared to postoperative application, and in comparison with placebo. Ninety patients were split randomly into three groups: group A received lornoxicam 60min before surgery and placebo 60min after surgery; group B received placebo 60min before surgery and lornoxicam 60min after surgery; group C received placebo 60min before surgery and placebo 60min after surgery. Postoperative pain was recorded on a visual analogue scale and on a numerical rating scale at 1, 2, 4, 6, 8, 12, and 24h after surgery. The patients recorded total dose of paracetamol intake during the 24h after the procedure. The efficacy of postoperative analgesia was greater in lornoxicam groups when compared to the placebo group; there was no difference between the two lornoxicam groups (A and B). Patients in group C took their first rescue analgesic dose earlier after surgery than patients in the two lornoxicam groups. The average dose of paracetamol taken in group C was 1000mg, while it was500 mg in the lornoxicam groups.

Effect of submucosal dexamethasone injection on postoperative pain, oedema, and trismus following mandibular third molar surgery: a prospective, randomized, double-blind clinical trial.

The aim of this study was to investigate the effect of the submucosal injection of 1ml dexamethasone (4mg/ml) on pain, swelling, and trismus following the extraction of retained lower third molars. Ninety patients (mean age 23.5 years) were split randomly into three equal study groups (30 patients in each): the 'before' group received dexamethasone 15min before surgery and placebo 15min after surgery; the 'after' group received placebo 15min before surgery and dexamethasone 15min after surgery; the 'placebo' group received placebo 15min before surgery and placebo 15min after surgery. Postoperative pain was recorded by the patients using a visual analogue scale, numerical rating scale, and the McGill Pain Questionnaire at 1, 2, 4, 6, 8, 12, and 24h after surgery. The patients also recorded the total number of analgesic doses consumed during the 24h after the procedure. Swelling (determined using linear measurements of the face) and trismus (determined through measurement of maximum mouth opening) were assessed at 48h, 72h, and 7 days following surgery. Better control of pain, swelling, and trismus was demonstrated for dexamethasone in comparison to placebo. Postoperative dexamethasone provided better pain control than preoperative dexamethasone. There was no difference in total rescue analgesic intake between the preoperative and postoperative dexamethasone groups.

Hyperkeratosis in human lower limb lymphedema: the effect of stagnant tissue fluid/lymph.

BACKGROUND: Hyperkeratosis of skin in lower limb lymphedema is one of the sequelae of tissue fluid/lymph (TF/L) stasis, but its mechanisms remain unknown. It is noteworthy, nonetheless, that human TF/L contains high levels of growth factors and cytokines, and may serve as the physiological environment for keratinocyte (KC) proliferation. OBJECTIVE: The aim of the study was to investigate the effect of human TF/L on human KC proliferation, differentiation and on the expression of epidermal stem cell markers on them. METHODS: KC were isolated from lymphedema and normal skin, and cultured for 1-14 days in TF/L with neutralized Interleukin 1ß, Interleukin 6, tumour necrosis factor α (TNF-α), keratinocyte growth factor (KGF) or tumour growth factor ß (TGF-ß). Alternatively, KC receptors for these factors were blocked. RESULTS: The number of KC cultured in TF/L was increased, as was the percentage of mitotic figures. There was a higher percentage of p63, CD29, Ki67, PCNA, CK6, CK17, CK16 and a lower of CK10, CK14, filaggrin and involucrin-positive KC. Neutralization of TF/L IL-1ß, IL-6, TNF-α and KGF as well as blockage of their receptors resulted in decreased percentage of mitotic KC. TGF-ß had a limited effect on KC proliferation. CONCLUSION: Hyperkeratosis in lymphedema may be the effect of a high concentration of cytokines in the stagnant TF/L tissue, but not because of presumed changes in the KC.

Transplantation of keratinocyte spore-like stem cells.

BACKGROUND: Wound granulation tissue should be covered by epidermal cells migrating from the basal layer of the epidermis or hair "bulge" of the wound edge. However, new epidermal islands are frequently formed on the granulation tissue remote from the wound edge. Thus, current theory of "bulge"-originating stem cells does not necessarily correspond to the histological pictures of the healing wound. We took imprints of a leg ulcer surface and found single dispersed, large nucleated cells, some of them in mitosis. These cells resembled those from epidermal spinosum layer. The question arouse as to whether these cells might be the "spore-like" stem cells creating epidermal island. We found similarly shaped cells among the keratinocyte preserved in pulverized sodium chloride as the only surviving population in culture and revealing enzymatic activity. The aim of this work was to study whether the population of human keratinocytes surviving sodium chloride preservation and transplanted to SCID mice may form epidermis. METHODS: The 12-month sodium chloride-preserved and cultured keratinocytes (KC) were transplanted to the wound on the dorsum of SCID mice for 14 and 21 days. RESULTS: Ninety-five percent of cultured KC were enzymatically active "large" cells; they did not express p63 and CD29 claimed as specific for stem cells, and they did not proliferate. Transplanted to the center of the wound, they formed small KC islands and became confluent after 14 days. CONCLUSIONS: The "large" epidermal keratinocytes survived the 12-month preservation in anhydrous sodium chloride. Transplanted to the wound, they formed epidermal islands of human phenotype. These cells may be the so-called "spore-like" stem cells.

Alternative method of oral administration by peanut butter pellet formulation results in target engagement of BACE1 and attenuation of gavage-induced stress responses in mice.

Development of novel therapeutic agents aimed at treating neurodegenerative disorders such as Alzheimer's and Parkinson's diseases require chronic and preferentially oral dosing in appropriate preclinical rodent models. Since many of these disease models involve transgenic mice that are frequently aged and fragile, the commonly used oro-gastric gavage method of drug administration often confounds measured outcomes due to repeated stress and high attrition rates caused by esophageal complications. We employed a novel drug formulation in a peanut butter (PB) pellet readily consumed by mice and compared the stress response as measured by plasma corticosterone levels relative to oral administration via traditional gavage. Acute gavage produced significant elevations in plasma corticosterone comparable to those observed in mice subjected to stress-induced hyperthermia. In contrast, corticosterone levels following consumption of PB pellets were similar to levels in naive mice and significantly lower than in mice subjected to traditional gavage. Following sub-chronic administration, corticosterone levels remained significantly higher in mice subjected to gavage, relative to mice administered PB pellets or naive controls. Furthermore, chronic 30day dosing of a BACE inhibitor administered via PB pellets to PSAPP mice resulted in expected plasma drug exposure and Aß40 lowering consistent with drug treatment demonstrating target engagement. Taken together, this alternative method of oral administration by drug formulated in PB pellets results in the expected pharmacokinetics and pharmacodynamics with attenuated stress levels, and is devoid of the detrimental effects of repetitive oral gavage.

Chronic lower limb wounds evoke systemic response of the lymphatic (immune) system.

Wound healing should not be considered as a process limited only to the damaged tissues. It is always accompanied by an intensive local immune response and in advanced stages, the systemic lymphatic (immune) structure. In this review we present evidence from our own studies as well as pertinent literature on the role of skin and subcutaneous tissue lymphatics at the wound site and of transport of antigens along with collecting afferent lymphatics to the lymph nodes. We also speculate the role of lymph nodes in raising cohorts of bacterial and own tissue antigen-specific lymphocytes and their participation in healing and not infrequently evoking uncontrolled chronic immune reaction causing a delay of healing. It is also speculated as to why there is a rapid response of lymph node cells to microbial antigens and tolerance to damaged-tissue-derived antigens occurs.

Long-lasting neuroprotection and neurological improvement in stroke models with new, potent and brain permeable inhibitors of poly(ADP-ribose) polymerase.

BACKGROUND AND PURPOSES: Thienyl-isoquinolone (TIQ-A) is a relatively potent PARP inhibitor able to reduce post-ischaemic neuronal death in vitro. Here we have studied, in different stroke models in vivo, the neuroprotective properties of DAMTIQ and HYDAMTIQ, two TIQ-A derivatives able to reach the brain and to inhibit PARP-1 and PARP-2. EXPERIMENTAL APPROACH: Studies were carried out in (i) transient (2 h) middle cerebral artery occlusion (tMCAO), (ii) permanent MCAO (pMCAO) and (iii) electrocoagulation of the distal portion of MCA in conjunction with transient (90 min) bilateral carotid occlusion (focal cortical ischaemia). KEY RESULTS: In male rats with tMCAO, HYDAMTIQ (0.1-10 mg·kg(-1)) injected i.p. three times, starting 4 h after MCAO, reduced infarct volumes by up to 70%, reduced the loss of body weight by up to 60% and attenuated the neurological impairment by up to 40%. In age-matched female rats, HYDAMTIQ also reduced brain damage. Protection, however, was less pronounced than in the male rats. In animals with pMCAO, HYDAMTIQ administered 30 min after MCAO reduced infarct volumes by approximately 40%. In animals with focal cortical ischaemia, HYDAMTIQ treatment decreased post-ischaemic accumulation of PAR (the product of PARP activity) and the presence of OX42-positive inflammatory cells in the ischaemic cortex. It also reduced sensorimotor deficits for up to 90 days after MCAO. CONCLUSION AND IMPLICATIONS: Our results show that HYDAMTIQ is a potent PARP inhibitor that conferred robust neuroprotection and long-lasting improvement of post-stroke neurological deficits.

Pathways of lymph and tissue fluid flow during intermittent pneumatic massage of lower limbs with obstructive lymphedema.

Questions remain on the use of sequential pneumatic compression including where does the fluid flow to and whether fluid can be moved to the non-swollen tissues of the hypogastrium and gluteal region? During pneumatic massage of the limb, we studied pathways of lymph and mobile tissue fluid flow using lymphoscintigraphy: a) from the calf and thigh across the inguinal region to the healthy non-swollen tissues of the hypogastrium and b) in the hypogastrium to the lateral and upper abdominal quadrants. To examine if there was effective fluid flow during pneumatic massage, plethysmographic flow measurements were also carried out. We demonstrated that: (i) pneumatic compression moved isotope in lymph remaining in functioning lymphatics and in tissue fluid in the interstitial space toward the inguinal region and femoral channel, (ii) there was no isotope crossing the inguinal crease or moving to the gluteal area, and (iii) isotope injected intradermally in the hypogastrium did not spread during manual massage to the upper and contralateral abdominal quadrants. In conclusion, intermittent pneumatic compression is effective in pushing mobile tissue fluid and relocating large fluid volumes toward the groin. However, the question that still remains is how to facilitate further flow toward the non-swollen tissues and thereby increase local absorption of fluid.

Influence of chronic caprine arthritis-encephalitis virus infection on the population of peripheral blood leukocytes.

The influence of caprine arthritis-encephalitis (CAE) virus infection on the population of peripheral blood leukocytes in goats was evaluated. For this purpose two groups of adult dairy female goats were formed. The experimental group consisted of 17 goats, which had been naturally infected for many years. The control group comprised 29 non-infected goats, which originated from CAE-free herd. All goats were clinically healthy. Whole blood was collected and tested in hematological analyzer and light microscope to assess the total number of leukocytes and the percentage of four leukocyte populations--neutrophils, eosinophils, monocytes and lymphocytes. Then, flow cytometry with monoclonal antibodies against several surface antigens (namely CD14, CD2, B-B2, CD4, CD8h, TCR-N6, WC1-N2 and WC1-N3) was performed to assess the proportion of lymphocyte subpopulations. Statistically significant differences (alpha < or = 0.01) were observed only in the subpopulations of T lymphocytes--percentage of all subpopulations were significantly higher in the group of seropositive goats. No statistically significant differences were revealed with respect to the total number of blood leukocytes, the average percentage of blood leukocyte populations and proportions of both T and B lymphocytes.

The effect of heterotypic infections of older horses with equine influenza virus type-2 on some clinical and immunological parameters.

Twelve horses, all of them 10 years old, were vaccinated intramuscularly on 0 and 28 days of the experiment with inactivated vaccine containing only antigens of A-equi-2/Miami/63. Another three unvaccinated horses, each at the age of 10 years, were the negative control group. One, ten-year-old horse was vaccinated with commercial inactivated vaccine containing both antigens of A-equi-2/Miami/63 as well as A-equi-1/Praha/56 as positive control. Three horses were challenged intranasally with homotypic strain of Miami/63, while six other were challenged with heterotypic strains--three with Suffolk/89 and three with Kentucky/86. Three horses vaccinated with vaccine containing only strain A-equi-2/Miami/63 were not challenged. In the group of three unvaccinated horses, each one was challenged intranasally with different strains studied in this experiment. The horse vaccinated with commercial vaccine was not challenged. Replication of each strain was done in chick embryos. During the experiment blood from horses was collected for hematological and immunological examinations (antigen-specific and antigen-nonspecific lymphocyte transformation tests, lymphocyte immunophenotyping, antigen-specific leukocyte migration inhibition test and hemagglutination inhibition test). The statistical analysis showed that the dynamics of lymphocyte immunological reactivity in horses vaccinated with inactivated vaccine containing antigens of A-equi-2/Miami/63 in response to further antigen stimulation (in vitro) was different comparing the homotypic or nearly homotypic challenging with Miami/63 and Suffolk/89 respectively, to the more heterotypic one with the strain Kentucky/86. In horses challenged with classical homotypic strain of Miami/63 no clinical signs were observed. These results confirm that the vaccine shall consist of the strains currently circulating in the horse population.

Influence of caprine arthritis-encephalitis virus infection on the activity of peripheral blood lymphocytes.

Non-specific lymphocyte transformation assay using phytohemagglutinin (PHA) as a mitogen was applied to evaluate influence of caprine arthritis-encephalitis virus (CAEV) infection on activity of peripheral blood lymphocytes. Animals were selected for the CAEV-infected and CAEV-non-infected groups according to the results of two serological surveys carried out at one year interval, with the use of an ELISA test. In goats which were not infected with CAEV, lymphocytes stimulation index (SI) revealed a high diversity of the results with an mean value equal to 5.86 (minimum = 0.45, maximum = 40.00, SD = 8.40). SI values for infected goats reached the average of 1.10 (minimum = 0.46, maximum = 1.85, SD = 0.26). The difference between the average lymphocyte stimulation indices was statistically highly significant in both groups (p = 0.002) which could be an evidence of CAEV infection influence on lymphocyte reactivity. Regarding ELISA test as a "golden standard" the application of lymphocyte transformation assay in diagnosis of CAEV infection was assessed. The ROC curve was drawn. The area under the curve was only 0.324, which indicates very low accuracy of this method and limits its use for the diagnosis of the disease.

Preemptive effect of ketoprofen on postoperative pain following third molar surgery. A prospective, randomized, double-blinded clinical trial.

The authors examined whether ketoprofen administered 60 min before surgical extraction of the lower wisdom teeth provides effective postsurgical analgesia and reduces rescue analgesic intake compared with ketoprofen administered 60 min after surgery or placebo. The 96 patients were placed into three groups: pre-group (ketoprofen 60 min preoperatively); post-group (ketoprofen 60 min postoperatively); and no-group (placebo). Study interventions had a significant effect on pain sensations in the 12 h after surgery. The initial onset of pain was significantly delayed only in the post-group. Pain intensity at the first onset of pain was significantly lower only in the post-group. Patients in the pre- and post-groups required significantly less rescue analgesic than those in the no-group. Ketoprofen administered after third molar surgery provides more effective pain control than ketoprofen administered before the surgery or placebo.

Where do lymph and tissue fluid accumulate in lymphedema of the lower limbs caused by obliteration of lymphatic collectors?

Obliteration of lymphatic collecting trunks of limbs by infective processes, trauma, oncologic surgery and irradiation bring about retention of lymph and tissue fluid in tissues. Knowledge as to where excess lymph is produced and accumulates as tissue fluid is indispensable for rational physical therapy. So far, this knowledge has been based on lymphoscintigraphic, ultrasonographic and MR images. None of these modalities provides distinct images of dilated lymphatics and fluid expanded tissue spaces in dermis, subcutis and muscles. Only anatomical dissection and histological processing of biopsy material can demonstrate the remnants of the lymphatic network and the sites of accumulation of mobile tissue fluid. We visualized and calculated the volume of the "tissue fluid and lymph" space in skin and subcutaneous tissue of foot, calf, and thigh in various stages of lymphedema, using special coloring techniques in specimens obtained during lymphatic microsurgical procedures or tissue debulking. When the collecting trunks were obliterated, lymph was present only in the subepidermal lymphatics, while mobile tissue fluid accumulated in the spontaneously formed spaces in the subcutaneous tissue, around small veins, and in the muscular fascia. Deformation of subcutaneous tissue by free fluid led to formation of interconnecting channels. In obstructive lymphedema caused by obliteration of collectors, lymph is present mainly in subepidermal lymphatics, and the bulk of stagnant tissue fluid accumulates in subcutis between fibrous septa and fat globules as well as above and underneath muscular fascia. These observations provide useful clues for designing pneumatic devices and rational manual lymphatic massage to move stagnant tissue fluid toward the non-swollen regions.

The influence of tissue fluid/lymph cytokines and growth factors on human keratinocyte proliferation and differentiation.

Cultured keratinocytes (KC) are needed for transplantation to the surface of large burn wounds and ulcers. They can be cultured in artificial media. However, the yield is always limited, viability is low, and proliferation and migration after grafting are slow. The question arose whether tissue fluid/lymph, which is a natural humoral environment for epidermal and dermal cells, contains cytokine(s) specifically regulating KC proliferation and could be used to culture large numbers of cells for transplantation. Culturing of skin keratinocytes in dermal tissue fluid/lymph containing keratinocyte growth factor, interleukin-1beta, interleukin-6, tumor necrosis factor-alpha, and transforming growth factor-beta revealed its strong stimulatory effect on the expression of p63 stem cell marker and proliferation but not differentiation of KC. Neutralizing these cytokines with antibodies resulted in decreased percentages of mitotic figures. None of the individual cytokines showed a dominant effect on proliferation. This observation suggests that either there may be other (so far undetected) specific cytokines or that the proliferation and differentiation of keratinocytes is an effect of the combined action of all investigated cytokines.

Some morphological, biochemical and immunological blood parameters in European bison with or without lesions in the digestive, respiratory, urinary and reproductive tracts.

Ninety-seven bison (39 males and 58 females) were culled out during three successive winter seasons. No infection with BHV-1 and BHV-5 or other viruses was detected by PCR and SN methods. Advanced pathological lesions observed in males exhibiting symptoms of posthitis/balanoposthitis are probably due to secondary infection by microorganisms present in the prepuce (Corynebacterium spp., Bacillus spp., Fusobacterium necrophorum, Staphylococcus spp. (coagulase negative) and Staphylococcus hyicus). Histopathological examinations of sections from prepuce and lips of pudendum showed the presence of Onchocercinae nematode (Onchocerca sp.). A comparison of biochemical and morphological blood parameters in the bison with or without lesions in the digestive (presence of parasites), respiratory, urinary and reproductive tracts revealed that the most variable parameter during reproductive organ disorder is the leukocyte count, while disorders in the urinary and digestive tracts coincided with the age of animals. In cases of digestive disorder most changes were observed in the number of neutrophils and concentration of urea. The number of red blood cells, monocytes, the level of alkaline phosphatase (AP) and hematocrit negatively correlated with age. Negative correlation was also observed between the number of neutrophils, level of creatinine and lymphocyte proliferation index. However, positive correlation was seen between erythrocyte sedimentation rate (Spearman's correlation coefficient), level of alanine aminotransferase (ALT) and protein, and age of the bison. Positive correlation was also observed between the lymphocyte count and lymphocyte proliferation index. The attempt to determine the reference values for blood biochemical and morphological parameters showed similarities to those of cattle.

Single-dose and multi-dose clindamycin therapy fails to demonstrate efficacy in preventing infectious and inflammatory complications in third molar surgery.

The goal of this study was to evaluate the efficacy of single- and multi-dose (5-day) clindamycin therapy for the prevention of inflammatory complications in patients undergoing lower third molar surgical extraction with bone removal. Patients who qualified for the prospective, randomized, double-masked, placebo-controlled trial were randomly divided into three groups: (1) single dose of oral clindamycin administered preoperatively (single-dose group); (2) clindamycin administered preoperatively with continued therapy for 5 days (5-day group); and (3) a placebo group. The following parameters were evaluated on the first, second and seventh days postsurgery: trismus, facial swelling, body temperature, lymphadenopathy, alveolar osteitis and subjective pain sensations. There were 86 patients (31 in the single-dose group, 28 in the 5-day group and 27 in the placebo group) enrolled in the study. There were no statistically significant differences in postoperative inflammatory complications in patients during the first and second days postsurgery. A statistically significant variation in body temperature was reported on the seventh day. Analysis of the postoperative analgesic intake did not show statistically significant differences between examined groups. Clindamycin applied in a single preoperative dose of 600 mg with or without subsequent 5-day therapy does not demonstrate efficacy in prophylaxis for postoperative inflammatory complications after third molar surgery.

The effectiveness of long-acting penicillin (penidur) in preventing recurrences of dermatolymphangioadenitis(DLA) and controlling skin, deep tissues, and lymph bacterial flora in patients with "filarial" lymphedema.

Dermatolymphangioadenitis (DLA) is a common and serious complication of so-called "filarial" and bacterial non-filarial lymphedema of the limb, affecting skin, lymphatics and lymph nodes. In our previous studies, we demonstrated that more than 60% of patients revealed presence of bacterial isolates in deep tissues, tissue fluid and lymph from the lymphedematous limbs. The question remained open whether elimination or suppression of bacteria dwelling in lymphedematous tissues by administration of low doses of penicillin for long time periods would prevent recurrence of DLA attacks. In this study, we retrospectively evaluated a self/community-selected group of patients with lymphedema of the lower limbs with respect to the efficacy of long-acting penicillin in preventing episodes of DLA. There were no microfilariae or anti-filarial antibodies detected in the investigated group. The questions we asked were: (a) how effective is the benzathine penicillin in preventing recurrences of DLA attacks and (b) how does its long-term administration influence the bacterial spectrum of leg skin, deep tissues, lymph and lymph nodes and sensitivity to antibiotics. Two randomly selected groups of patients, receiving and not receiving penicillin during the same period of time, were compared. Evidently lower recurrence rate of DLA was observed in the treated group (p < 0.002). There was increased prevalence of cocci and gram-positive bacilli with a concomitant decrease of gram-negative bacilli on the foot and calf skin surface. Simultaneously, decreased prevalence of gram-positive cocci and gram-negative bacilli isolates in limb deep tissues and lymph was seen. No resistance to penicillin and other tested antibiotics developed in isolates from the skin surface, deep tissues and lymph. We conclude that long-lasting penicillin is effective in preventing recurrent DLA attacks.

Helicobacter pylori infection in patients with haemophilia in Poland: prevalence and risk of upper gastrointestinal bleeding.

Infection with Helicobacter pylori is the main aetiological factor for erosive gastritis and duodenal or gastric peptic ulcers often complicated with life-threatening bleeding in patients with coagulation disorders. The aim of this prospective study was to evaluate the prevalence of Helicobacter pylori infection in haemophilia patients, and to assess the risk of gastrointestinal bleeding associated with this infection. From 2000 to 2002, 146 patients with haemophilia (129, haemophilia A; 13, haemophilia B), mean age, 39.9 years (+/-7.3), were investigated for H. pylori infection using IgG and IgA latex serological test. The control group included 100 men with no coagulation disorders, mean age, 40.9 years (+/-9.2). For 72 (49.3%) patients with haemophilia and 39 controls (39.0%) serological tests were positive indicating the presence of H. pylori infection (P =0.1112). A history of gastrointestinal bleeding was reported in 46 patients (31.5%) with haemophilia and in two control group patients (2.0%) (P < 0.0001). Gastrointestinal bleeding was significantly more frequent in patients with haemophilia infected with H. pylori (33/46; 71.7%) than in patients with no H. pylori infection (13/46; 28.3%; P = 0.0002). In conclusion, the prevalence of H. pylori infection in haemophilic patients in Poland is comparable with that in patients with no coagulation disorders. Helicobacter pylori infection is a risk factor for duodenal and gastric ulcer bleeding in haemophilia patients. In view of the high frequency of upper gastrointestinal bleeding associated with H. pylori infection, we believe that screening and eradication therapy are appropriate in haemophilia patients.

Virulence-associated traits in avian Escherichia coli: comparison between isolates from colibacillosis-affected and clinically healthy layer flocks.

Colibacillosis appears to be of increasing importance in layer flocks. The aim of this study was to determine characteristics of avian pathogenic Escherichia coli associated with the occurrence of colibacillosis outbreaks at flock level. Forty E. coli strains originating from layers from healthy flocks ('control isolates'), consisting of 25 caecal and 15 extra-intestinal isolates, were compared with 40 strains isolated from layers originating from colibacillosis-affected flocks ('outbreak isolates'), consisting of 20 caecal and 20 extra-intestinal isolates. The examined characteristics were adhesins, invasivity in T84 cell culture, serum resistance, iron uptake, colicin production, and toxinogenicity. The following traits were significantly more often detected in the outbreak isolates than in the control isolates: tsh, iss, iucA, iutA, irp2, fyuA, iroC, cvaC, colicin and colicin V production. A comparison of the extra-intestinal outbreak isolates and the caecal control isolates yielded the same results as when the caecal isolates, extra-intestinal isolates and total number of isolates of the outbreak and the control group were compared. When comparing the caecal and extra-intestinal isolates within the control and within the outbreak group, no significant differences were detected. The O78 and O2 groups showed significant differences with other O-types and NT strains for prevalence of most of the same characteristics. The combination of type 1 fimbriae, tsh, serum resistance, iss, traT, iucA, fyuA, iroC and colicin or colicin V production was significantly more often present in extra-intestinal outbreak isolates than in extra-intestinal control isolates. Only the combination of serum resistance, fyuA and colicin production was present in all outbreak isolates, with a significantly lower prevalence in the control isolates. None of the characteristics or combinations examined were exclusive to the outbreak isolates.

Targeted destruction of normal and cancer cells through lutropin/choriogonadotropin receptors using Hecate-betaCG conjugate.

A recent approach to cancer treatment is destruction of malignant and non-malignant tumors by hormonally targeted lytic peptides. The presence of lutropin/choriogonadotropin (LH/CG) receptors has been confirmed in several cancer cells (e.g. breast, ovarian, and prostate). In a series of experiments conducted in vitro, we have used a conjugate of the 23-amino acid lytic peptide Hecate and a 15-amino acid segment of beta-chain of CG. To test the hypothesis that Hecate-betaCG selectively destroys porcine granulosa and luteal cells, and Leydig cancer cell line (BLT-1) possessing LH/CG receptors, the conjugate was added to culture media at different concentrations of 0.5 to 10 micro M. Spleen cells and late passage of granulosa cancer cell line (KK-1) not-possessing LH/CG receptors were used as controls. The toxicity of Hecate-betaCG conjugate was concentration-dependent in all cell types but different among various cells. The toxicity of the conjugate to treated cells was closely correlated with the number of LH/CG receptors per cell. At low concentration (1 micro M), Hecate-betaCG was more cytotoxic to cells bearing LH/CG receptors than to controls (p < 0.01). In contrast to cells possessing LH/CG receptors, cancer cell line KK-1 and spleen cells were sensitive only at concentration of 5 micro M (p < 0.001). We conclude that Hecate-betaCG selectively kills cells expressing LH/CG receptors; its toxicity is dependent on the number of binding sites for LH/CG.