High prevalence of epidermodysplasia verruciformis-associated human papillomavirus DNA in actinic keratoses of the immunocompetent population

Skin cancers in both immunosuppressed and immunocompetent populations are associated with epidermodysplasia verruciformis human papillomavirus (EV-HPV) DNA. However, little is known about the prevalence of EV-HPVs in actinic keratoses in immunocompetent individuals. Actinic keratoses from 114 patients were classified as low-grade (n=76) or high-grade (n=38) according to the extent of histological atypia. HPV DNA was amplified from 54 frozen and 60 paraffin-embedded biopsy specimens by nested polymerase chain reaction (PCR) with several consensus and type-specific primers. PCR products were sequenced for typing. These results were compared with HPV detection in skin cancers (n=20) and Bowens disease (n=18). A broad spectrum of EV-HPV types including oncogenic HPV5 and HPV8 and partially characterized sequences were detected in actinic keratoses and cutaneous cancers. In actinic keratoses a higher prevalence of EV-HPV DNA was found in frozen tissues than in formalin-fixed tissues (85% vs 67%). There was no difference between the low- and high-grade actinic keratoses either in terms of EV-HPV DNA prevalence or the results of serological study using HPV8 virus-like particles. The detection rate of EV-HPVs was lower in skin cancers and Bowens disease. This would suggest involvement of EV-HPVs in the early stages of cutaneous oncogenesis.Dr. Fuchs has died since this article was completed.     

Tick-borne encephalitis in north-eastern Poland in 1997-2001: a retrospective study

The aim of this study was to characterize the epidemiology and clinical features of tick-borne encephalitis in north-eastern Poland. Clinical and epidemiological data were analysed of patients hospitalized with the diagnosis in the Department of the Infectious Diseases and Neuroinfections of the Medical University in Bialystok in 1997-2001. Tick-borne encephalitis was diagnosed in 152 patients: 51 (34%) presented with meningitis, 89 (59%) with meningoencephalitis and 12 (8%) with meningoencephalomyelitis. Headache (84%) and fever (81%) were the most common symptoms. Meningeal signs were present in 137 patients (90%). Most common neurological abnormalities were: Oppenheim and Babinski signs (74 patients, 49%), ataxia (37, 24%), impaired consciousness (37, 24%) and pareses (16, 10%). Of patients examined, 146 (96%) had raised pleiocytosis, frequently accompanied by high cerebrospinal fluid protein concentration (90%), raised erythrocyte sedimentation rate (65%), peripheral blood leucocytosis (26%) and increased aminotransferase activity (16%). There was only 1 forest worker among the patients. Tick-borne encephalitis remains common in north-eastern Poland but, possibly because of effective vaccination, it has virtually disappeared among forest employees. The diagnosis appears difficult in some cases, as meningeal signs may not be present and laboratory findings may not be suggestive of a viral infection.     

Identification of carotenoids in ovarian tissue in women

Epidemiological and clinical studies have revealed that vitamin A and its derivatives (carotenoids and retinoids) can reduce the risk of ovarian tumours and may have a role in the metabolism of patients with ovarian cancer. The aim of the study was identification and quantitative assessment of carotenoids found in nature, mainly of provitamin A group, in the tissue material obtained from patients with different lesions of the ovaries. Material for analysis was obtained from 100 women, aged 16-74, operated on for ovarian tumours in the Department of Gynaecology. Carotenoid pigments were separated using column chromatography, thin-layer chromatography and high-performance liquid chromatography. In the tissue material subjected to analysis, 14 carotenoids were identified, including provitamin A carotenoids; beta-carotene, beta-cryptoxanthin, echinenone and hydroxyechinenone. alpha-carotene was not found. In the whole group of pathological lesions, the total carotenoid content was relatively low (mean 1.717 microg/g tissue) and the mean content of provitamin A carotenoids was 17.28%. These results are similar to results obtained in the group of normal ovarian tissue. In the group of benign mucinous tumours (1.042 microg/g tissue) and tumours in the thecoma-fibroma group (1.328 microg/g tissue) and dysgerminoma group (1.279 microg/g tissue), the total carotenoid content was lower. Only in the endometriosis group was this value higher (2.185 microg/g tissue). Epoxy carotenoids; lutein epoxide, violaxanthin and mutatoxanthin were predominant (in %). Irrespective of histological classification, beta-carotene, beta-cryptoxanthin, lutein, lutein epoxide, violaxanthin and mutatoxanthin were identified in all tissue examined. Antheraxanthin was isolated in all tissue except for normal ovarian tissue, serous malignant and mucinous benign and malignant tumours, endometrioid malignant tumours, dermoid cysts, corpus luteum cysts and simple cysts. Hydroxyechinenone was isolated sporadically. Only in one case was capsanthin isolated. Carotenoids act as chemopreventive agents, irrespective of whether they are finally transformed into vitamin A, and may represent a potentially powerful alternative to present chemotherapeutic approaches to the treatment of ovarian cancer.     

Structure-activity relationships for a series of peptidomimetic antimicrobial prodrugs containing glutamine analogues

Synthetic glutamine analogues such as N3-(4-methoxyfumaroyl)-l-2,3-diaminopropanoic acid (FMDP) inhibit purified glucosamine-6-phosphate synthase, an intracellular enzyme that is essential for microbial cell wall synthesis, but they are inactive against intact organisms because they cannot enter the cell. However, when the analogues are linked to a peptide they can be actively transported, and FMDP peptidomimetics show broad-spectrum antimicrobial activity. To characterize this process in more detail, the antibacterial activities of various synthetic peptidomimetics containing glutamine analogues have been determined against isogenic strains of Escherichia coli in which one or more of its three peptide transporters Dpp, Opp and Tpp have been mutated. In addition, their affinities for DppA and OppA, the binding-protein components of the transporters, have been measured. In general, antibacterial activities against the various transport mutants correlated with binding to DppA and OppA. Xaa-FMDP compounds have greater activities than FMDP-Xaa analogues. To explore structure-activity relationships for the peptidomimetics, molecular modelling was used to determine the conformational forms they adopt in solution. The relative bioactivities of the peptidomimetics correlated with the percentage of conformers that had backbone torsions matching those previously defined for the molecular recognition templates of the peptide transporters. However, the large size of the N-terminal residue in the FMDP-Xaa analogues appears to interfere with transport and thus to limit antibacterial activity. Overall, the results provide the structural rationale for the identification in silico of analogues with optimal bioactivities, which decreases the need for extensive chemical syntheses and testing.     

Crystallinity of human pineal calcospherulites

Summary Crystallinity of mineral in human pineal calcospherulites was determined by electron spin resonance spectrometry after irradiation of the samples with gamma rays in a⁶⁰Co-source. The radiation-induced stable paramagnetic centers in the crystalline lattice of hydroxyapatite crystals were used as a marker of the crystalline fraction and related to the total mineral content. The crystallinity of pineal sand is higher than that of compact bone. The numerical value of the crystallinity coefficient depends on both the average crystal size of hydroxyapatite and the percentage of the crystalline fraction in the total amount of mineral. Literature data show that the average size of hydroxyapatite crystals in pineal sand are smaller than in bone tissue. It is, therefore, concluded that the higher crystallinity of pineal acervuli is due to the lower percentage of the submicrocrystalline fraction in their mineral.     

AbetaPP-overexpression and proteasome inhibition increase alphaB-crystallin in cultured human muscle: relevance to inclusion-body myositis

Amyloid-beta precursor protein (AbetaPP) and its fragment amyloid-beta (Abeta) are increased in s-IBM muscle fibers and appear to play an important role in the pathogenic cascade. alphaB-Crystallin (alphaBC) was shown immunohistochemically to be accumulated in s-IBM muscle fibers, but the stressor(s) influencing alphaBC accumulation was not identified. We now demonstrate, using our experimental IBM model based on genetic overexpression of AbetaPP into cultured normal human muscle fibers, that: (1) AbetaPP overexpression increased alphaBC 3.7-fold (p=0.025); (2) additional inhibition of proteasome with epoxomicin increased alphaBC 7-fold (p=0.002); and (3) alphaBC physically associated with AbetaPP and Abeta oligomers. We also show that in biopsied s-IBM muscle fibers, alphaBC was similarly increased 3-fold (p=0.025) and physically associated with AbetaPP and Abeta oligomers. We propose that increased AbetaPP is a stressor increasing alphaBC expression in s-IBM muscle fibers. Determining the consequences of alphaBC association with Abeta oligomers could have clinical therapeutic relevance.     

Cathepsin D and carcino-embryonic antigen in serum, urine and tissues of colon adenocarcinoma patients

BACKGROUND/AIMS: Application of neoplastic markers in early diagnosis of colorectal carcinoma has brought fresh hope to millions of sufferers. However such a marker, distinctive for this particular carcinoma and allowing its detection at a sufficiently early stage of development has not yet been found. Cathepsin D (CD) is lysosomal aspartyl proteinase. It is a component of a proteolytic cascade participating actively in neoplastic invasion as well as in metastasis formation. Carcino-embryonic antigen (CEA) is a useful marker in oncological diagnostics of colorectal cancer. CEA undergoes expression in all kinds of adenocarcinoma and is found both intercellularly and extracellularly. High concentrations of CEA in the blood serum confirm neoplastic changes in the digestive tract with high probability. The objective of this study has been to evaluate CD activity in the blood serum, urine and tumor tissues as well as in the colon biopsies which were not changed macroscopically and CEA concentration in the serum of colon adenocarcinoma, considering the extent of spread of cancer (TNM), the grade of the differentiation of cancer cell (G) as well as the tumor size. The possibility of application of CD along with CEA as markers of colon adenocarcinoma has also been examined. METHODOLOGY: The examination included the serum and urine of 21 patients as well as 12 tissues biopsies with histopathologically confirmed colon adenocarcinoma. The reference group for the blood and urine comprised of 17 healthy controls, and for the colon adenocarcinoma tissues- samples collected from 14 people from the sites most distant from the resected tumor on the boundaries which were free of cancer cells. Activity of CD in the blood serum, urine as well as tissues was determined with a modified Greczaniuk et al. method and expressed by the amount of released tyrosine as the concentration of the activity in nmolTyr/mL/6h, whereas the specific activity was expressed in nmol Tyr/mg of protein /6h. The specific activity of CD in the urine was expressed in nmol Tyr/mg of creatinine/6h. CEA concentration in the blood serum was determined by the immunoenzymatic method (MEIA) on Axym Abbot Analyzer and was expressed in ng/mL. The protein concentration was determined by the Lowry method, and the results were expressed in mg/mL. The creatinine concentration in the urine was determined by the Jaffe method (without deproteinization) and was expressed in mg/100mL. RESULTS: CD activity was increased in the blood serum (p < 0.0001) and tissues (p = 0.022) of colon adenocarcinoma patients in comparison to the reference group. CD specific activity (Tyr/mg of protein/6h) was significantly increased in serum but decreased in the urine (p < 0.0001) whereas the specific activity of CD (nmol Tyr/mg of creatinine/6h) was increased in the urine (p = 0.0001). CD specific activity has tendency to increase in colon adenocarcinoma tissues (p = 0.441) as compared to the reference group. By examining data in regard to TNM clinical-histopathological classification, G and the tumor size, it could be concluded that CD activity in serum and urine in colon adenocarcinoma patients depends on progress of cancer in which CD activity increases with TNM. A statistically significant increase in CEA concentration was found in the serum of colon adenocarcinoma patients, which was almost threefold higher than the in reference group. No significant differences in CEA concentration were found depending on TNM, G and tumor size. CONCLUSIONS: The results of this study suggest that examination of CD activity and CEA concentration in serum, as well as CD activity in the urine, might be used in oncological diagnostics of colon adenocarcinoma.     

Management of Anastomotic Leak: Lessons Learned from a Large Colon and Rectal Surgery Training Program

Background

Anastomotic leak is a dreaded surgical complication that can lead to significant morbidity and mortality. Despite its prevalence, there is no consensus on the management of anastomotic leak. This study aimed to review the management of anastomotic leak in the Division of Colon and Rectal Surgery at two institutions.

Methods

This is a retrospective review of all anastomotic leaks occurring after surgery in the Division of Colon and Rectal Surgery at two teaching institutions during 1997–2008.

Results

Altogether, 103 leaks occurred in 1,707 anastomoses (6 %), with a median time to diagnosis of 20 days (2–1,400 days). The 90-day mortality rate was 3 %. The majority of cases were managed nonoperatively (73 %), and the majority of leaks were from an extraperitoneal anastomosis (67 %). Success (i.e., radiographic demonstration of a healed leak, restored gastrointestinal continuity) occurred in 54 % of operatively managed leaks and 57 % of nonoperatively managed leaks (56 % overall). Operative management differed by leak location. In 91 % of patients with intraperitoneal leaks, the anastomosis was resected. In 76 % of patients with extraperitoneal leaks, diversion and drainage alone was performed without manipulating the anastomosis. Nonoperative management was successful for 57 % of extraperitoneal leaks and 58 % of intraperitoneal leaks. There was no significant difference in the success rates based on type of management (operative/nonoperative) for either extraperitoneal or intraperitoneal leaks.

Conclusions

Anastomotic leak continues to result in patient morbidity and mortality. Its diverse presentation requires tailoring management to the patient. Nonoperative and operative treatments are viable options for intraperitoneal and extraperitoneal leaks based on patient presentation.     

Sealing of Vessels Larger Than 7 Millimeters Using Enseal in Porcine Aorta

Background:

The Enseal (Ethicon Endo-Surgery, Blue Ash, Ohio) tissue-sealing device has proven efficacy for ligation of vessels <7 mm in diameter, even with significant supraphysiologic bursting pressures. We aimed to evaluate the safety of Enseal in porcine vessels >7 mm.

Materials and Methods:

The lumbar aortas of pigs that were euthanized for unrelated procedures were harvested. A 5- to 6-cm segment of aorta was sealed using the Enseal device. The opposite end was attached to a pressure-testing device to measure pressures at leak or bursting. The bivariate Pearson correlation was used to determine the relationship between diameter and bursting pressure. One-way analysis of variance was used to determine differences between the groups of vessels on the basis of their diameter.

Results:

Ninety samples of 5-cm aorta segments were used to assess bursting pressure. The median diameter was 14 mm (range, 7–18) and bursting pressure was 85 mm Hg (range, 24–650). The Pearson test showed a negative correlation between vessel diameter and bursting pressure (P = .25). One-way analysis of variance did not show any significant difference between vessel diameters grouped by size (P = .517), and neither did the Scheffe post hoc test when comparing diameter with bursting pressure; 31% of specimens failed to seal.

Conclusions:

Bursting pressures are low and inconsistent after tissue sealing with the Enseal device in porcine vessels >7 mm. These vessels also demonstrated a higher rate of failure to seal. The histologic results of the aorta segments (ie, a low collagen-elastin ratio) may be the cause of the low bursting pressures.