A new laboratory device with mathematically based positioning of a frozen tissue block facilitating precise sectioning of large specimens.

Mohs micrographic surgery (MMS) is a treatment method aiming at thorough, personalized eradication of skin cancers by mean of staged excision of tissues surrounding the tumor with complete (100%) histopathological examination of their margins. In many MMS laboratories, the excised tissue is divided, shaped, frozen in a cryostat with a heat extractor and positioned manually (with the block on the object disc) in an articulated cryostat chuck during cutting. However, these activities may be difficult, time-consuming and associated with the risk of imprecise tissue sectioning. Development of a laboratory device allowing for processing of large tissue specimens, with the function of mechanical, mathematically steered positioning of the tissue block surface directly to the microtome knife cutting place, eliminating the need for manual adjustment. The prototype device was designed and manufactured. Its functioning was tested on 513 histological slides produced during 212 operations of skin cancers using MMS. The depth of the first complete sections and the diameter of sections were measured. Complete sections were obtained at an average depth of 81.60 m (min. 20 m, max. 180 m, SD = 29.15), whereas the average diameter of sections was 18.11 mm (min. 4 mm, max. 42 mm, SD = 9.10). The histological processing of large specimens with mathematically based positioning of the tissue surface in relation to the cryotome knife cutting plane is precise, fast and easy. The device can be useful in those MMS centers which continue to employ manual setting of the cryostat chuck or share the cryostat with other users, which prevents fixing the chuck position (including large hospital settings). It may also be helpful in centers using a cryostat with a fixed chuck, for the correction of minimal inaccuracies of its preset position.

Acitretin treatment for hidradenitis suppurativa: a prospective series of 17 patients.

BACKGROUND: The management of hidradenitis suppurativa (HS) is still challenging, and new treatment methods are necessary. Acitretin seems to be a promising agent in HS management; however, literature data are limited. OBJECTIVES: To investigate the clinical efficacy of acitretin monotherapy in 17 patients with long-standing and recalcitrant HS. Simultaneously, an assessment of quality of life was carried out. METHODS: The study was conducted with a group of 17 patients with HS. Patients were treated with acitretin for up to 9 months and examined at baseline, after 1 month, and then every 3 months from baseline. The clinical extent of disease severity was measured with the HS Severity Index (HSSI), Hurley scoring system, Physician's Global Assessment and Dermatology Life Quality Index (DLQI). RESULTS: Nine patients (53%) finished the whole 9 months of acitretin treatment. The mean acitretin dose was 0·56 ± 0·08 mg kg(-1) daily. A significant improvement of clinical manifestation (HSSI, DLQI) was observed after only 1 month of therapy, and further improvement was recorded during the next few months. Overall, eight subjects (47%) fulfilled the criteria for response (HSSI ≥ 50% reduction from baseline). The dropout rate was 47%, due mostly to drug ineffectiveness and adverse events. Discontinuation of treatment resulted in deterioration or relapse of HS 2-8 months after acitretin cessation, in all but one patient. CONCLUSIONS: HS treatment with acitretin seems to be a promising method of disease management. However, due to the high daily dosage, its usage may be limited.

Carbon materials as new nanovehicles in hot-melt drug deposition.

The application of commercially available carbon materials (nanotubes and porous carbons) for the preparation of drug delivery systems is studied. We used two types of carbon nanotubes (CNT) and two activated carbons as potential materials in so-called hot-melt drug deposition (HMDD). The materials were first studied using Raman spectroscopy. Paracetamol was chosen as a model drug. The performed thermal analysis, kinetics, and adsorption-desorption studies revealed that nanoaggregates are formed between carbon nanotubes. In contrast, in pores of activated carbon we do not observe this process and the drug adsorption phenomenon mechanism is simply the filling of small pores. The formation of nanoaggregates was confirmed by the results of GCMC (grand canonical Monte Carlo) simulations and the study of the surface area on nitrogen adsorption-desorption isotherms. The application of carbon nanotubes in HMDD offers the possibility of controlling the rate of drug delivery. Performed MTT tests of nanotubes and drug-loaded nanotubes show that the observed decrease in cell viability number is caused by the influence of the cytostatic properties of nanotubes-they inhibit the proliferation of cells. The carbon nanotubes studied in this paper are essentially nontoxic.

Secondary intention healing in skin surgery: our own experience and expanded indications in hidradenitis suppurativa, rhinophyma and non-melanoma skin cancers.

BACKGROUND: There are many situations in integument surgery in which secondary intention healing (SIH) may bring results comparable or even superior to those obtained with primary intention healing. OBJECTIVE: To present our own methods of employing SIH in surgical treatment of different skin lesions. Moreover, to delineate the extended indications for SIH as a first-line method of wound treatment. METHODS: Between 1999 and 2009, we conducted 236 operations followed by SIH in the group of 195 patients. Two independent investigators and the patients carried out post-operative assessment (at 12 months) of both cosmetic and functional results of operations (data for 142 patients). Moreover, at 3 and 12 months, patients were asked for self-evaluation of the outcomes of surgical procedures using the visual analogue scale (VAS). RESULTS: The most frequent indications for SIH were defects after the excision of non-melanoma skin cancers and hidradenitis suppurativa lesions. Depending on localization, the mean extent of the post-operative wounds prepared for SIH ranged from 0.8to 85.7 cm(2). The healing lasted 2-12 weeks. The most important factor was the size of the wound (r = 0.59; P < 0.001). The assessment at 12 months revealed that cosmetic/functional outcomes were evaluated as 'good' or 'excellent' in 89/83% and 72/70% cases, according to patients and physicians respectively. VAS showed that evaluation scores at 3 months post-surgery were significantly lower than at 12 months (P < 0.0001). CONCLUSION: SIH is a safe and efficient form of treatment for various post-operative skin defects. In certain areas of the body, SIH shows favourable cosmetic and functional effects. In these cases, it may even be regarded as the method of choice.

Role of base-excision repair in the treatment of childhood acute lymphoblastic leukaemia with 6-mercaptopurine and high doses of methotrexate.

Methotrexate (MTX) and 6-mercaptopurine (6MP) are the most commonly used drugs in the therapy of childhood acute lymphoblastic leukaemia (ALL). The main genotoxic effect of MTX resulting from inhibition of thymidylate synthase is mis-incorporation of uracil into DNA, which is considered essential for the effectiveness of the Protocol M in ALL IC BFM 2002/EURO LB 2002 regimens. In this study, we investigated the level of basal and induced DNA damage as well as the effectiveness of DNA repair in lymphocytes of children with ALL at four time-points during therapy with MTX and 6MP. To assess DNA damage and the efficacy of DNA repair we used the modified alkaline comet assay with uracil DNA glycosylase (Udg) and endonuclease III (EndoIII). In addition, we examined the induction of apoptosis in the lymphocytes of the patients during treatment. Finally, we compared the activity of base-excision repair (BER), involved in removal of both uracil and oxidized bases from DNA in lymphocytes of children with ALL and lymphocytes of healthy children. BER efficiency was estimated in an in vitro assay with cellular extracts and plasmid substrates of heteroduplex DNA with an AP-site. Our results indicate that there is a significant decrease in the efficacy of DNA repair associated with an increased level of uracil in DNA and induction of apoptosis during therapy. Moreover, it was found that the BER capacity was decreased in the lymphocytes of ALL patients in contrast to that in lymphocytes of healthy children. Thus, we suggest that an impairment of the BER pathway may play a role in the pathogenesis and therapy of childhood ALL.

XPD common variants and their association with melanoma and breast cancer risk.

There are suggestions in the literature that common variants in the XPD gene may be associated with an altered risk of melanoma and breast cancer. To establish if the XPD common variants Asp312Asn and Lys751Gln are associated with an increased melanoma or breast cancer risk we performed an association study based on genotyping 426 unselected patients with malignant melanoma (MM) and 1830 consecutive breast cancer cases and compared the results to 1262 geographically matched newborns, 621 adults from the region of Szczecin (unselected for age and cancer family history), 421 healthy adults age- and sex-matched with the melanoma cases and 511 healthy controls matched with the breast cancer patients from the region of Szczecin. Additionally we examined the prevalence of three additional XPD variants, Gly156Gly, Leu485Pro and Arg112His amongst the 421 unselected melanoma patients. All of the variants when evaluated singularly were found not to be associated either with melanoma or breast cancer risk in younger or older patients. A modest association was observed with breast cancer risk when the Lys751Gln_CC/Asp312Asn_AA genotype (OR=1.5, p<0.05) segregated together. Individuals harboring the Lys751Gln_CC/Gly156Gly_CC genotype were significantly over-represented among late-onset melanoma cases (OR=1.7, p<0.05). The results of analyses of linkage disequilibrium and haplotype frequency support the thesis that a combination of at least two SNPs (Lys751Gln_CC/Gly156Gly_CC or Lys751Gln_CC/Asp312Asn_AA) inherited as a haplotype was associated with disease. These two pairs of SNPs could therefore be regarded as a single hereditary unit that would have a very small probability of being disrupted by recombination. Additional studies are required to determine whether these particular changes can be associated with an increased risk of other malignancies at different sites of origin.

Selected haemostatic factors in carotid bifurcation plaques of patients undergoing carotid endarterectomy.

OBJECTIVE: To determine the concentration of selected haemostatic factors (HFs): thrombin-antithrombin complexes (TAT), antithrombin (AT), tissue plasminogen activator (t-PA), plasminogen activator inhibitor type 1 (PAI-1) and D-dimers in carotid bifurcation plaques and to compare plaque composition in different subgroups of patients (mainly those with symptomatic and asymptomatic carotid stenosis). MATERIALS AND METHODS: Thirty-eight consecutive patients (20 symptomatic, 18 asymptomatic) undergoing carotid endarterectomy were enrolled in the study. The concentration of selected HFs in carotid plaques was measured using mainly enzyme immunoassay (ELISA). Simultaneously, the concentration of HFs in plasma was also obtained. RESULTS: Symptomatic plaques contained significantly more TAT complexes (p=0.03). AT was found only in nine out of 38 carotid plaques and was present mainly in symptomatic carotid plaques (n=8/9)(p<0.006). No significant differences were found between symptomatic and asymptomatic carotid plaques with respect to t-PA, PAI-1 and D-dimers concentration. There was an increased concentration of TAT (p<0.001), t-PA (p<0.02) and D-dimers (p<0.02) in carotid plaques of diabetic patients. Patients with coexisting intermittent claudication had elevated levels of D-dimers in carotid plaques (p<0.02). The only positive correlation was demonstrated between the concentration of AT in plasma and carotid plaques (R=0.76; p=0.02). CONCLUSIONS: All the evaluated HFs are the components of a carotid plaque. Symptomatic patients have increased concentration of TAT complexes in a carotid plaque. The symptomatic carotid plaque contains AT more frequently, which correlates positively with AT plasma levels. The most marked changes in the carotid plaque haemostatic composition (expressed by elevated levels of TAT, t-PA and D-dimers) have diabetic patients.

Brooke-Spiegler syndrome.

The Brooke-Spiegler syndrome is an autosomal dominant one characterized by cylindromas, trichoepitheliomas and occasionally spiradenomas. Within a given family, some members may have cylindromas whereas others may have trichoepitheliomas or both. We describe the coexistence of trichoepithelioma papulosum multiplex (also known as epithelioma adenoides cysticum of Brooke) and cylindromas in a 30-year-old man, and discuss the relationship between these two autosomal dominant syndromes.

[Morphologic studies of the reaction of the subcutaneous connective tissue in rats to implantation of porous ceramics]. / Badania morfologiczne reakcji tkanki lacznej podskórnej szczura na wszczep ceramiki porowatej.

In the authors carried out testing of the porous ceramics Al2O3 biocompatibility. Comminuted ceramics with an average graining 3.5 were inserted subcutaneously in suspension of physiological salt in rats. The animals were killed after 7, 15, 30, 60 and 180 days. Histologic testing of the skin together with the subcutis in the place of the ceramics' powder injection as well as testing of the internal organs was carried out. It was observed that already after 30 days the connective tissue reaction became stable and did not change until the end of the experiment. A connective tissular capsule 120-160 mu depth of collagenous fibre and some fibroblasts of the correct structure came into existence around the inserted ceramics' powder. In the course of the whole experiment neither macrophages nor great foreign bodies were observed. Accumulation of the ceramics' molecules and RES reactions were not observed in the internal organs. The tested ceramics in characterized by high tissular biocompatibility.

Application of porous bioceramic in experimental therapy of bone injuries. III. Dynamics of the callus development at the site of porous bioceramic implantation. Morphological, histochemical and histoenzymological studies.

In this work the biocompatibility of porous bioceramic implanted to the rabbit femoral bone was studied. The animals were killed 3, 6, 9, 14, 18 and 30 days after implantation and the callus with surrounding periosteum from the site of implant was taken for the studies. Morphological investigations of the callus were carried out up to the 30th day of healing of the bone tissue. Moreover, acid mucopolysaccharides level and activity of enzymes (acid and alkaline phosphatase, aminopeptidase, non-specific alpha-esterase, adenosine triphosphatase and succinate dehydrogenase) were studied up to the 18 day of the callus development. The results show that after bioceramic implantation, morphology of particular stages of the callus development, behaviour of acid mucopolysaccharides as well as localization and activity of enzymes are the same as in the normal healing process of the injured bone tissue. After 30 days total union of the mature bone tissue with bioceramic was established. We conclude that porous bioceramic satisfies the requirements for biomedical materials and may be safely used in the treatment of certain bone system diseases in humans.

Application of porous bioceramic in experimental therapy of bone injuries. II. Histoenzymological studies in the control animals.

In this work the activity of enzymes participating in the bone repair process was studied in rabbits on the 3rd, 6th, 9th, 14th and 18th day after mechanical injury of the bone tissue. The enzymes under study included acid and alkaline phosphatase, adenosine triphosphatase, non-specific alpha-esterase and succinate dehydrogenase.

Application of porous bioceramic in experimental therapy of bone injuries. I. Morphological and histochemical studies in the control animals.

The process of healing of the rabbits' bone tissue after mechanical injury was studied. The femoral bone was injured by means of oscillating saw. The animals were killed 3, 6, 9, 14, 18 and 30 days after operation and the callus with the surrounding periosteum was taken for morphological examination. Besides, acid mucopolysaccharides level and aminopeptidase activity in the callus (up to the 18th day of healing) were determined. The time of total bone union was established for 30 days.