Is Shock Wave Application Effective on Various Chronic Wounds in the Geriatric Population? Preliminary Clinical Study.

Purpose: Extracorporeal shock wave therapy (ESWT) is a beneficial adjunct modality for chronic wounds. Limited research has been conducted on pressure ulcers (PUs), while the majority of studies have focused on diabetic foot ulcers (DFUs) and venous leg ulcers (VLUs). This study aimed to evaluate the short-term effects of radial ESWT in older adults with chronic wounds. Patients and Methods: This study involved a total of 31 wounds: PUs (n=22), VLUs (n=7), and DFUs (n=2). A single radial ESWT was performed with 300 + 100 shocks per cm2, pressure of 2.5 bar, energy of 0.15 mJ/mm2, and frequency of 5 Hz. Assessments using digital planimetry and clinical methods, utilizing the Wound Bed Score (WBS) and the Bates-Jansen Wound Assessment Tool (BWAT) were performed before the radial ESWT application (M0) and one week after (M1). Results: A significant wound decrease in planimetry was noted (pre-ESWT vs post-ESWT), with wound area from 9.4 cm2 to 6.2 cm2, length from 6.4 cm to 3.9 cm, and width from 2.8 cm to 2.1 cm (p<0.001). Additionally, a substantial clinical improvement was noted in both the WBS with a 31.25% increase and the BWAT with a 20.00% increase (p<0.001). It was also found a significant correlation between the planimetric and clinical outcomes for both tools: WBS (r=-0.446, p=0.012) and BWAT (r=0.327, p=0.073). Conclusion: The ESWT application yields substantial immediate clinical effects that support the healing of chronic wounds in older adults. Even a single ESWT session can prove to be clinically effective and beneficial in the management of chronic wounds.

Can Shockwave Treatment Elicit a Molecular Response to Enhance Clinical Outcomes in Pressure Ulcers? The SHOck Waves in wouNds Project.

Wound healing requires the coordinated interaction of dermis cells, the proper deposition of extracellular matrix, re-epithelialization, and angiogenesis. Extracorporeal shock wave (ESW) is a promising therapeutic modality for chronic wounds. This study determined the biological mechanisms activated under ESW, facilitating the healing of pressure ulcers (PUs). A group of 10 patients with PUs received two sessions of radial ESW (300 + 100 pulses, 2.5 bars, 0.15 mJ/mm2, 5 Hz). Histomorphological and immunocytochemical assessments were performed on tissue sections obtained from the wound edges before the ESW (M0) and after the first (M1) and second (M2) ESW. The proliferation index of keratinocytes and fibroblasts (Ki-67), the micro-vessels' density (CD31), and the number of myofibroblasts (α-SMA) were evaluated. The involvement of the yes-associated protein (YAP1) in sensing mechanical strain, and whether the nuclear localization of YAP1, was shown. The increased proliferative activity of epidermal cells and skin fibroblasts and the increased number of myofibroblasts, often visible as integrated cell bands, were also demonstrated as an effect of wound exposure to an ESW. The results indicate that the major skin cells, keratinocytes, and fibroblasts are mechanosensitive. They intensify proliferation and extracellular matrix remodeling in response to mechanical stress. A significant improvement in clinical wound parameters was also observed.

The Immediate Clinical Effects Following a Single Radial Shock Wave Therapy in Pressure Ulcers: A Preliminary Randomized Controlled Trial of The SHOWN Project.

Objective: Recent preliminary reports indicate that extracorporeal shock wave therapy (ESWT) might be useful for chronic wounds, especially venous leg ulcers and diabetic foot ulcers. However, there is limited evidence for the utility and safety of ESWT in pressure ulcers (PUs). Therefore, this randomized controlled trial (RCT) aimed to assess immediate planimetric and clinical effects following a single radial ESWT session in PUs. Approach: A group of 40 patients with PUs was randomly assigned into 2 groups: active ESWT (n = 20), which underwent a single treatment with radial ESWT (300 + 100 impulses/1 cm2, 2.5 bars, 0.15 mJ/mm2, and 5 Hz) and placebo ESWT (n = 20), which was exposed to sham-radial ESWT. All patients continued standard wound care procedures. The planimetric assessment and clinical outcomes using Wound Bed Score (WBS) and Bates-Jansen Wound Assessment Tool (BWAT) were assessed before (M0) and after ESWT sessions (M1). Results: There was a significant planimetric enhancement observed after active ESWT reported as a reduction in all metric parameters: wound area from 11.51 to 8.09 cm2 (p < 0.001), wound length from 4.97 to 4.41 cm (p < 0.001), and wound width from 3.15 to 2.49 cm (p < 0.0001). Also, there was a significant beneficial clinical improvement observed with a WBS as an increased score from 3.85 to 9.65 points (p < 0.001) and with the BWAT as a decreased score from 45.45 to 30.70 points (p < 0.001). In turn, a regression in the placebo ESWT group was observed in all studied outcomes. Innovation: This study is the first RCT to provide the positive and immediate clinical effects of radial ESWT in promoting the healing of PUs. Conclusion: This preliminary RCT showed that even a single session of ESWT is a promising and clinically effective modality in managing PUs. However, there is still limited data regarding the usefulness of ESWT in PUs, and further studies are in demand.

Effect of Radial Extracorporeal Shock Wave Therapy on Pain Intensity, Functional Efficiency, and Postural Control Parameters in Patients with Chronic Low Back Pain: A Randomized Clinical Trial.

Low back pain (LBP) is the leading cause of disability worldwide, placing a significant economic burden on healthcare systems. Radial extracorporeal shock wave therapy (rESWT) is useful in the rehabilitation of orthopedic diseases; however, there is still limited evidence for patients with LBP. The aim of this study was to assess the effect of rESWT on pain level, functional efficiency, and parameters of postural control in patients with LBP. Participants were randomized into group A (n = 20) treated with rESWT and group B (n = 20) treated with sham rESWT (placebo). Both groups received conventional physiotherapy, including core stability exercises. The following tests were performed: the Laitinen Pain Scale (LPS), the Roland-Morris Questionnaire (RMQ), the original Schober Test (OST), and a stabilometric platform for the assessment of postural sway, including total sway path (TSP). We found that the analgesic effect was higher after rESWT, especially in the follow-up's (p < 0.05). Similar results were found for functional efficiency and range of motion (p < 0.05). The improved posture stability in placebo group B was not statistically significant (p > 0.05). The stabilometric parameters in group A were still gradually improved and statistically significant, even in follow-ups (p < 0.05). In conclusion, the rESWT had a significant effect on the reduction of pain and the improvement of functional condition compared to a conventional physiotherapy program. Also, rESWT with core stability exercises led to significant improvements in postural sway compared with conventional physiotherapy in patients with LBP.

Shock Waves as a Treatment Modality for Spasticity Reduction and Recovery Improvement in Post-Stroke Adults - Current Evidence and Qualitative Systematic Review.

PURPOSE: This systematic review examines intervention studies using extracorporeal shock wave therapy (ESWT) application in post-stroke muscle spasticity with particular emphasis on the comparison of two different types of radial (rESWT) and focused shock waves (fESWT). METHODS: PubMed, PEDro, Scopus, and EBSCOhost databases were systematically searched. Studies published between the years 2000 and 2019 in the impact factor journals and available in the English full-text version were eligible for inclusion. All qualified articles were classified in terms of their scientific reliability and methodological quality using the PEDro criteria. The PRISMA guidelines were followed and the registration on the PROSPERO database was done. RESULTS: A total of 17 articles were reviewed of a total sample of 303 patients (age: 57.87±10.45 years and duration of stroke: 40.49±25.63 months) who were treated with ESWT. Recent data confirm both a subjective (spasticity, pain, and functioning) and objective (range of motion, postural control, muscular endurance, muscle tone, and muscle elasticity) improvements for post-stroke spasticity. The mean difference showing clinical improvement was: ∆=34.45% of grade for fESWT and ∆=34.97% for rESWT that gives a slightly better effect of rESWT (∆=0.52%) for spasticity (p<0.05), and ∆=38.83% of angular degrees for fESWT and ∆=32.26% for rESWT that determines the more beneficial effect of fESWT (∆=6.57%) for range of motion (p<0.05), and ∆=18.32% for fESWT and ∆=22.27% for rESWT that gives a slightly better effect of rESWT (∆=3.95%) for alpha motor neuron excitability (p<0.05). The mean PEDro score was 4.70±2.5 points for fESWT and 5.71±2.21 points for rESWT, thus an overall quality of evidence grade of moderate ("fair" for fESWT and "good" for rESWT). Three studies in fESWT and four in rESWT obtained Sackett's grading system's highest Level 1 of evidence. CONCLUSION: The studies affirm the effectiveness of ESWT in reducing muscle spasticity and improving motor recovery after stroke.

The Effectiveness Of Radial Extracorporeal Shock Wave Therapy In Patients With Chronic Low Back Pain: A Prospective, Randomized, Single-Blinded Pilot Study.

PURPOSE: This prospective, randomized and single-blinded study assesses the influence of radial extracorporeal shock wave therapy (rESWT) in patients with low back pain (LBP). METHODS: A total of 52 patients with LBP were enrolled in the study, out of which a homogeneous group of 40 patients with mean age of 53.45±4.9 years was included. Patients were randomized into group A (n=20) treated with rESWT (2000 pulses; 2.5 bars; 5 Hz, 7 mins) performed twice a week for five weeks (10 sessions) and stabilization training, as well as group B (n=20) treated with sham rESWT and stabilization training. To analyze the therapeutic progress, the following tests were performed (before and after therapy; 1 and 3 months follow-up) to assess pain and functional efficiency: (1) Visual Analog Scale (VAS), (2) Laitinen Pain Scale (LPS), and (3) Oswestry Disability Index (ODI). RESULTS: The control group had a statistically significant advantage over the rESWT group (4.4 vs. 3.1 points on the VAS; p=0.039). However, in long-term observations, group A gradually experienced more pain relief than group B (2.7 vs. 3.5 points, p>0.05, at one month and 2.0 vs. 4.4 points at three months after treatment; p<0.0001). Similar findings can be seen in the analysis of changes in pain sensations measured with the LPS. The functional state (ODI) was better in rESWT group, especially in follow-up observation (9.3 vs. 14.6 points, p=0.033, at one month and 9.3 vs. 17.8 points, p=0.004, at three months after treatment). CONCLUSION: The rESWT combined with stabilization training is particularly effective in the long-term and achieves a stable beneficial effect for patients with LBP. The use of rESWT has a significant long-term influence on the reduction of pain and the improvement of the general functional state in relation to the conventional motor improvement program.

Photobiomodulation using high- or low-level laser irradiations in patients with lumbar disc degenerative changes: disappointing outcomes and remarks.

BACKGROUND: Laser therapy seems to be a beneficial physical agent for chronic low back pain (LBP), and it is commonly used in the clinical rehabilitation practice. However, there are still no indisputable and clearly defined protocols and practical guidelines, and further, the methodology of the previous reports leaves many unsatisfied and raises some reservations. OBJECTIVE: The aim of this study was to evaluate the effectiveness of low-level laser therapy (LLLT) and high-intensity laser therapy (HILT) in patients with lumbar disc degenerative changes based on the analysis of the short- and long-term results and in comparison with the placebo effect. DESIGN: This study was a prospective and placebo-controlled clinical trial. MATERIALS AND METHODS: A group of 68 participants were qualified for the therapy and were assigned to four comparative groups in the order they volunteered: HILT of 1,064 nm, 60 J/cm2, 10 minutes (HILT); sham (HILT placebo); LLLT of 785 nm, 8 J/cm2, 8 minutes; and sham (LLLT placebo). The following tests were used to assess the effectiveness of treatment: 1) the visual analogue scale; 2) the Laitinen Questionnaire Indicators of Pain; 3) the Oswestry Disability Index; 4) the Roland-Morris Disability Questionnaire; 5) Lasegue test; and 6) Schober's test. All measurements were carried out before and after irradiations (3 weeks) and in follow-ups (1 and 3 months). RESULTS: After applying verum or placebo laser irradiation, therapeutic progress was observed in all comparative groups; however, no statistically significant differences were observed among the procedures. CONCLUSION: The high- and low-energy laser therapy methods used in the present article are ineffective in relation to patients with lumbar disc degenerative changes in both the short- and long-term perspectives and do not show a significant advantage over the placebo effect.

Effect of laser therapy on expression of angio- and fibrogenic factors, and cytokine concentrations during the healing process of human pressure ulcers.

Objective: To evaluate the effect of laser irradiation at different wavelengths on the expression of selected growth factors and inflammatory mediators at particular stages of the wound healing process. Methods: Sixty-seven patients were recruited, treated, and analyzed (group A - 940 nm: 17 patients; group B - 808 nm: 18 patients; group C - 658 nm: 16 patients; group D - sham therapy: 17 patients). Patients received a basic treatment, including repositioning and mobilization, air pressure mattress and bed support surfaces, wound cleansing and drug therapy. Additionally, patients received laser therapy once a day, 5 times a week for 1 month in use of a semiconductor lasers (GaAlAs) which emitted a continuous radiation emission at separate wavelengths of 940 nm (group A), 808 nm (group B) and 658 nm (group C). In group D (sham therapy), laser therapy was applied in the same manner, but the device was off during each session (only the applicator was switched on to scan pressure ulcers using none coherent red visible light). Results: The positive changes in the measured serum (IL-2, IL-6 and TNF-α) and wound tissue (TNF-α, VEGF and TGFß1) parameters appeared to be connected only with the wavelength of 658 nm. The significant change in pro-inflammatory mediator levels [interleukin 2 (IL-2) with p=0.008 and interleukin 6 (IL-6) with p=0.016] was noticed after two weeks of laser therapy. In the other groups, the inflammation was also reduced, but the process was not as marked as in group C. Similarly, in the case of tumor necrosis factor (TNF-α) concentration, where after two weeks of treatment with irradiation at a wavelength of 658 nm, a rapid suppression was observed (p=0.001), whereas in the other groups, these results were much slower and not as obvious. Interestingly, again in the case of group C, the change in TNF-α concentration in wound tissue was most intensive (≈75% reduction), whereas the changes in other groups were not as obvious (≈50% reduction). After irradiation (658 nm), the VEGF expression increased significantly within the first two weeks, and then it decreased and maintained a stable level. In contrast, the TGFß1 activity remained level, but always higher in comparison to other groups Conclusions: The effective healing of pressure ulcers is connected with laser irradiation at a wavelength of 658 nm. We believe that this effect is related to the inhibition of inflammatory processes in the wound and stimulation of angiogenesis and fibroblast proliferation at this specific radiation (based both on concentration of interleukins and TNF-α serum level and VEGF, TGFß1, TNF-α activities in wound biopsies). Laser therapy at wavelengths of 940 and 808 nm does not significantly affect the above-mentioned repair processes, which explains its low effectiveness in the treatment of pressure ulcers.

Mechanoregulation of Wound Healing and Skin Homeostasis.

Basic and clinical studies on mechanobiology of cells and tissues point to the importance of mechanical forces in the process of skin regeneration and wound healing. These studies result in the development of new therapies that use mechanical force which supports effective healing. A better understanding of mechanobiology will make it possible to develop biomaterials with appropriate physical and chemical properties used to treat poorly healing wounds. In addition, it will make it possible to design devices precisely controlling wound mechanics and to individualize a therapy depending on the type, size, and anatomical location of the wound in specific patients, which will increase the clinical efficiency of the therapy. Linking mechanobiology with the science of biomaterials and nanotechnology will enable in the near future precise interference in abnormal cell signaling responsible for the proliferation, differentiation, cell death, and restoration of the biological balance. The objective of this study is to point to the importance of mechanobiology in regeneration of skin damage and wound healing. The study describes the influence of rigidity of extracellular matrix and special restrictions on cell physiology. The study also defines how and what mechanical changes influence tissue regeneration and wound healing. The influence of mechanical signals in the process of proliferation, differentiation, and skin regeneration is tagged in the study.

Nuclear location of tumor suppressor protein maspin inhibits proliferation of breast cancer cells without affecting proliferation of normal epithelial cells.

BACKGROUND: Maspin, which is classified as a tumor suppressor protein, is downregulated in many types of cancer. Several studies have suggested potential anti-proliferative activity of maspin as well as sensitizing activity of maspin for therapeutic cytotoxic agents in breast cancer tissue culture and animal models. All of the experimental data gathered so far have been based on studies with maspin localized cytoplasmically, while maspin in breast cancer tumor cells may be located in the cytoplasm, nucleus or both. In this study, the effect of maspin cytoplasmic and nuclear location and expression level on breast cancer proliferation and patient survival was studied. METHODS: Tissue sections from 166 patients with invasive ductal breast cancer were stained by immunohistochemistry for maspin and Ki-67 protein. The localization and expression level of maspin were correlated with estimated patient overall survival and percent of Ki-67-positive cells. In further studies, we created constructs for transient transfection of maspin into breast cancer cells with targeted cytoplasmic and nuclear location. We analyzed the effect of maspin location in normal epithelial cell line MCF10A and three breast cancer cell lines - MCF-7, MDA-MB-231 and SKBR-3 - by immunofluorescence and proliferation assay. RESULTS: We observed a strong positive correlation between moderate and high nuclear maspin level and survival of patients. Moreover, a statistically significant negative relationship was observed between nuclear maspin and Ki-67 expression in patients with invasive ductal breast cancer. Spearman's correlation analysis showed a negative correlation between level of maspin localized in nucleus and percentage of Ki-67 positive cells. No such differences were observed in cells with cytoplasmic maspin. We found a strong correlation between nuclear maspin and loss of Ki-67 protein in breast cancer cell lines, while there was no effect in normal epithelial cells from breast. The anti-proliferative effect of nuclear maspin on breast cancer cells was statistically significant in comparison to cytoplasmic maspin. CONCLUSIONS: Our results suggest that nuclear maspin localization may be a prognostic factor in breast cancer and may have a strong therapeutic potential in gene therapy. Moreover, these data provide a new insight into the role of cytoplasmic and nuclear fractions of maspin in breast cancer.

Embryonic and adult isoforms of XLAP2 form microdomains associated with chromatin and the nuclear envelope.

Laminin-associated polypeptide 2 (LAP2) proteins are alternatively spliced products of a single gene; they belong to the LEM domain family and, in mammals, locate to the nuclear envelope (NE) and nuclear lamina. Isoforms lacking the transmembrane domain also locate to the nucleoplasm. We used new specific antibodies against the N-terminal domain of Xenopus LAP2 to perform immunoprecipitation, identification and localization studies during Xenopus development. By immunoprecipitation and mass spectrometry (LC/MS/MS), we identified the embryonic isoform XLAP2γ, which was downregulated during development similarly to XLAP2ω. Embryonic isoforms XLAP2ω and XLAP2γ were located in close association with chromatin up to the blastula stage. Later in development, both embryonic isoforms and the adult isoform XLAP2ß were localized in a similar way at the NE. All isoforms colocalized with lamin B2/B3 during development, whereas XLAP2ß was colocalized with lamin B2 and apparently with the F/G repeat nucleoporins throughout the cell cycle in adult tissues and culture cells. XLAP2ß was localized in clusters on chromatin, both at the NE and inside the nucleus. Embryonic isoforms were also localized in clusters at the NE of oocytes. Our results suggest that XLAP2 isoforms participate in the maintenance and anchoring of chromatin domains to the NE and in the formation of lamin B microdomains.

Antiproliferative and pro-apoptotic effects of quercetin on human pancreatic carcinoma cell lines EPP85-181P and EPP85-181RDB.

Polyphenols are present in several edible plants and for many years induce high interest mainly due to their antioxidative and anti-inflammatory influence. At present, numerous studies are conducted on antineoplastic effects of the compounds. One of most effective biopolyphenols involves the flavonol quercetin. Our studies aimed at evaluation of antiproliferative and pro-apoptotic effects of quercetin alone and in combinations with daunorubicin on cells of human pancreatic carcinoma lines. The experiments were conducted on two cell lines, sensitive to daunorubicin EPP85-181P line, and its resistant variant EPP85-181RDB. Effect of studied substances on cell proliferation was detected using sulphorhodamine B (SRB) protein staining method. Apoptotic damage was estimated using comet and TUNEL techniques. Our data demonstrated that quercetin exerted cytotoxic action on cells of the both neoplastic cell lines in concentration-dependent manner. In the case of EPP85-181RDB cell line, quercetin seemed to sensitize resistant cells to daunorubicin. In parallel, the effect of both substances on the sensitive cell line was synergistic. Results of the studies confirmed that quercetin may probably break resistance of neoplastic cells to chemotherapy. On the other side, studied flavonol augmented action of cytostatic drug in case of sensitive tumour cells what suggest, that it might allow to decrease dosage of cytostatic drugs and reduce negative side effects of the treatment.

Quercetin as a potential modulator of P-glycoprotein expression and function in cells of human pancreatic carcinoma line resistant to daunorubicin.

P-glycoprotein (P-gp) is one of the ABC transporters responsible for the resistance of several tumours to successful chemotherapy. Numerous agents are capable of interfering with the P-gp-mediated export of drugs but unfortunately most of them produce serious side effects. Some plant polyphenols, including the flavonol quercetin (Q), manifest anti-neoplastic activity mainly due to their influence on cell cycle control and apoptosis. Reports are also available which show that Q may intensify action of cytostatic drugs and suppress the multidrug resistance (MDR) phenomenon. The study aimed at determination if Q sensitizes cells resistant to daunorubicin (DB) through its effect on P-gp expression and action. The experiments were conducted on two cell lines of human pancreatic carcinoma, resistant to DB EPP85-181RDB and sensitive EPP85-181P as a comparison. Cells of both lines were exposed to selected concentrations of Q and DB, and then membranous expression of P-gp and its transport function were examined. The influence on expression of gene for P-gp (ABCB1) was also investigated. Results of the studies confirmed that Q affects expression and function of P-gp in a concentration-dependent manner. Moreover it decreased expression of ABCB1. Thus, Q may be considered as a potential modulator of P-gp.

Influence of pressure on the endothelium of the saphenous vein coronary artery bypass graft.

The aim of this study was to evaluate the influence of pressure applied while assessing the graft's tightness on the expression of adhesion molecules. Another goal was to find a correlation between the type of fluid (heparynized blood or saline) used during preparation of the conduit and the expression of the adhesion molecules. Saphenous vein fragments were obtained from 48 patients who had undergone coronary artery surgery. Expression of the following particles was evaluated: CD 31, ICAM 1, VCAM 1 and P-selectin. Expression of the CD 31 molecule was described as a percentage of the inner surface of the vessel showing positive immunocytochemical reaction. Expression of the remaining molecules (ICAM 1, VCAM 1, P-selectin) was assessed as the percentage of the surface, determined by CD 31 positive reaction. The expression of the adhesion molecules (ICAM 1, VCAM 1, P-selectin) was higher in the fragments of the vein exposed to pressure. In reference to VCAM 1 the difference, as compared with the control group, was: 250% in the fragments infused with blood and 270% in the fragments infused with saline, respectively. The differences for the ICAM 1 were approximately 300% in both experimental groups and 450% for the P-selectin with subtle differences between the two experimental groups. The loss of the endothelial surface (determined by the expression of the CD 31 antigen) was similar in the specimens flushed either with blood or saline, which indicates that the major cause of damage of the endothelium is influence of pressure on the conduit's wall. Mechanical widening of vessels results in the increased expression of the adhesion molecules on the surface of the endothelial cells, and, as a consequence, leads to rise in the leukocyte adhesion and loss of the functional properties of the transplanted veins.

Maspin and c-erbB-2 expression in correlation with microvessel density in invasive ductal breast cancer.

Maspin is a unique member of the serpin family involved in regulation of cell migration, apoptosis and angiogenesis in breast and prostate cancers. In this study maspin expression in comparison with c-erbB-2 (HER2/neu) oncogene expression and microvessel density was investigated. The examined material included specimens of primary invasive ductal breast cancer derived from 69 patients. They were analyzed immunocytochemically to assess maspin and c-erbB-2 expression, as well as microvessel density using endothelium marker CD31. In the studied cancers, maspin expression in cancer cells was detected in more than half of the cases (50.73%). Although statistically insignificant (p=0.27), maspin expression showed decreasing tendency with the increase of tumor grade. C-erbB-2 oncogene expression was observed in 78.26% of the examined cancers. Statistically significant positive correlation was found between c-erbB-2 expression and tumor grade (p<0.005). Analysis of the dependence between maspin and c-erbB-2 expression exhibited statistically significant inverse correlation (p<0.001). Mean microvessel density (MVD) of the studied cancers was 71.64 (SD=19.36). MVD decreased with the increase of maspin expression, whereas in the cases showing c-erbB-2 overexpression MVD was clearly higher. Both correlations were statistically significant (p<0.005). In conclusion, it could be stated that increase in maspin expression is associated with weaker expression of c-erbB-2 oncogene and lower microvessel density, which implies a significant role of maspin in tumor biology. However, the exact mechanism of maspin action (including its potential role in angiogenesis), as well as the assessment of its prognostic significance in breast cancer require further studies.

Malignant myoepithelioma of the salivary gland--an untypical clinical course.

A case of malignant tumor developing from myoepithelial cells (malignant myoepithelioma) is presented. The primary focus was located in the region of the left submandibular salivary gland. A relapse and metastases were disclosed in the same salivary gland, in the lung and the left breast. Immunohistochemical studies demonstrated positive reactions for S-100 protein, cytokeratins, smooth muscle actin, vimentin, GFAP protein, p53 protein and Ki-67 antigen, and allowed for establishing the final histopathological diagnosis of malignant myoepithelioma.

Evaluation of apoptosis, proliferation intensity and metallothionein (MT) expression in comparison with selected clinicopathological variables in primary adenocarcinomas of the large intestine.

Tumour growth and expansion are the result of proliferative activity and the capacity to eliminate cells by apoptosis and/or necrosis. The present study was aimed at comparing the apoptosis and proliferation intensity in cells of adenocarcinomas of the large intestine with the expression of metallothionein (MT), the grade of the tumour and the depth to which the tumour infiltrated the intestinal wall. The TUNEL technique and immunocytochemical reactions (expression of caspase-3, Ki-67, MT) were used to detect apoptosis. The results demonstrated augmented levels of all the variables examined, positively correlated with grade of malignancy, G, and with the depth of intestinal wall infiltration by the tumour cells. The testing of apoptosis, proliferation and MT expression may prove useful in the appraisal of the growth and progression of primary adenocarcinomas in the large intestine.

The 35 kDa acid metallophosphatase of the frog Rana esculenta liver: studies on its cellular localization and protein phosphatase activity.

The cellular localization of the 35 kDa, low molecular mass acid metallophosphatase (LMW AcPase) from the frog (Rana esculenta) liver and its activity towards P-Ser and P-Tyr phosphorylated peptides were studied. This enzyme was localized to the cytoplasm of hepatocytes but did not appear in other cells of liver tissue (endothelium, macrophages, blood cells). This LMW AcPase does not display activity towards (32)P-phosphorylase a under conditions standard for the enzymes of PPP family. Proteins containing P-Ser: rabbit (32)P-phosphorylasea and phosvitin are hydrolysed only at acidic pH and are poor substrates for this enzyme. The frog AcPase is not inhibited by okadaic acid and F(-) ions, the Ser/Thr protein phosphatase inhibitors. Moreover, the frog enzyme does not cross-react with specific antisera directed against N-terminal fragment of human PP2A and C-terminal conserved fragment of the eukaryotic PP2A catalytic subunits. These results exclude LMW AcPase from belonging to Ser/Thr protein phosphatases: PP1c or PP2Ac. In addition to P-Tyr, this enzyme hydrolyses efficiently at acidic pH P-Tyr phosphorylated peptides (hirudin and gastrin fragments). K(m) value for the hirudin fragment (7.55 +/- 1.59 x 10(-6) M) is 2-3 orders of magnitude lower in comparison with other substrates tested. The enzyme is inhibited competitively by typical inhibitors of protein tyrosine phosphatases (PTPases): sodium orthovanadate, molybdate and tungstate. These results may suggest that the LMW AcPase of frog liver can act as PTPase in vivo. A different cellular localization and different response to inhibition by tetrahedral oxyanions (molybdate, vanadate and tungstate) provide further evidence that LMW AcPase of frog liver is distinct from the mammalian tartrate-resistant acid phosphatases.

Prognostic value of immunocytochemical determination of parathyroid hormone-related peptide expression in cells of mammary ductal carcinoma. Analysis of 7 years of the disease course.

Parathyroid hormone-related peptide (PTHrP) participates in the development of humoral hypercalcaemia of malignancy. The peptide is thought to affect growth and differentiation of normal and neoplastic cells. The present study aimed at evaluation of the relationship between survival time and development of distant metastases in patients with ductal mammary carcinoma on the one hand and PTHrP expression on the other. Immunocytochemical reactions using mouse monoclonal (clone 212-10.7) anti-PTHrP (38-64) antibodies were performed in paraffin sections originating from 47 patients with ductal mammary carcinoma. Expression of the protein was quantified employing a scale, considering the number of positive cells and intensity of the reaction (immunoreactive score, IRS). Survival time of the patients, determined during the course of a 7-year observation was also analysed. The obtained results demonstrated a relationship between intensity of PTHrP expression and the survival time. Patients with high expression of PTHrP (IRS>6) manifested longer survival than patients with lower PTHrP expression (IRS< or =6; Cox'es F test, P<0.05). Moreover, in the group with the lower PTHrP expression, a negative relationship was detected between expression of the protein and the survival time (Cox'es model, P<0.05). No relationship was detected between PTHrP expression and the development of distant metastases.

Role of exogenous melatonin in reducing the nephrotoxic effect of daunorubicin and doxorubicin in the rat.

The aim of these studies was to examine the nephroprotective effect of melatonin following the anthracycline administration [daunorubicin (DNR); doxorubicin (DOX)] in rats. Application of these drugs in chemotherapy is limited because of their cardiotoxicity and nephrotoxicity. Rats of the Buffalo strain were divided into groups according to the cytostatic drug used, its dose and sequence of administration [DNR or DOX single (i.v.) dose of 10 mg/kg b.w., i.e. acute intoxication and 3 mg/kg b.w. (i.v.) weekly for 3 wk, subchronic intoxication]. Melatonin was administered subcutaneously before and after every injection of a cytostatic drug at a dose of 10 mg/kg b.w. The severity of renal alterations was examined both biochemically [levels of lipid peroxidation markers, malonyldialdehyde (MDA) and 4-hydroxyalkenals (4-HDA)], or histologically. A statistically significant decrease in renal damage was noted after melatonin administration to acutely or subchronically intoxicated DNR-treated and DOX-treated rats. Biochemical assays revealed significant decreases in MDA and 4-HDA levels following application of melatonin during subchronic DNR or DOX intoxication. In summary, melatonin was found to exert a protective effect on the kidney, which was particularly evident after subchronic DOX and DNR intoxication, using both histological or biochemical methods.