A New Exopolysaccharide from a Wood-Decaying Fungus Spongipellis borealis for a Wide Range of Biotechnological Applications.

Fungi are a unique natural resource rich in polysaccharides, proteins, and other components. Polysaccharides are considered one of the most important bioactive components in fungi. Increasing numbers of studies have confirmed that fungal polysaccharides have various biological activities. Given these facts, the main aim of this investigation was to carry out isolation, identification, and structural characterisation of a new polysaccharide (EPS) derived from laboratory-cultured vegetative mycelium of a new Spongipellis borealis strain isolated from the environment. The examination of monosaccharides in the EPS demonstrated that the isolated biopolymer was composed mainly of glucose, galactose, and mannose monomers. The analysis of the methylation of the studied polymer indicated that it contained mainly terminal, →3)-linked, →4)-linked, and →2,4)-linked hexoses. The effect of fungal polysaccharides on S. borealis proteolytic enzymes (pepsin, trypsin, and pycnoporopepsin) and laccase activity was determined for the first time. Incubation of the enzyme preparation and EPS showed an influence of EPS on the stability of these enzymes, compared to the control values (without EPS).

Effect of bisphosphonates on selected markers of bone turnover in patients after total knee arthroplasty.

PURPOSE: Osteoporosis is a problem for many patients after total knee arthroplasty (TKA). The aseptic loosening of the prosthesis is also a significant problem. Therefore, in these patients, bisphosphonates (BPs) are used that, by influencing the level of bone turnover markers, reduce the risk of osteoporotic fractures and aseptic revisions in TKA. The purpose of the study was to assess whether the Pamifos® present in bone cement has any effect on the level of selected bone turnover markers and cytokines in patients after total knee arthroplasty. METHODS: The study group consisted of 30 women with degenerative changes of the knee joint, whose total knee prosthesis was stabilized with cement enriched with Pamifos®. The control group consisted of 30 women treated for degenerative changes of the knee joint without the use of bisphosphonate-enriched cement for prosthetic stabilization. RESULTS: In the study group, we found a decrease in tumour necrosis factor (TNF-α) levels 12 weeks after surgery, whereas the control group experienced an almost twofold increase in TNF-α level. The concentration of OPG, a natural RANKL antagonist, was highest in patients of the study group six weeks after surgery and was four times higher compared to the control group. Statistically significant differences were found in the RANKL level (P < 0.05). In the control group, there was a continuous increase in RANKL concentration from the first to the 12th week after surgery. The highest level of RANKL in patients of the study group was found six weeks after the surgery, and 12 weeks after knee arthroplasty, it was significantly lower. It was found that the concentration of osteocalcin (OC) in the study group was the lowest three weeks after the surgery, then it increased and remained at a similar level after 12 weeks. The concentrations of selected cytokines (IL-1ß, IL-2, IL-6, IL-10, IL-17AF) also showed statistically significant differences. CONCLUSIONS: The BP-stimulated increase in the level of OPG and the decrease in the level of RANKL, as well as the impact on the level of the analyzed interleukins in the bone microenvironment, may be an important element of the mechanisms limiting bone resorption. Therefore, the use of BP-enriched cement implants appears to be justified.

Thromboelastometric Analysis of Anticancer Cerrena unicolor Subfractions Reveal Their Potential as Fibrin Glue Drug Carrier Enhancers.

In this study, the influence of two subfractions (with previously proven anti-cancer properties) isolated from wood rot fungus Cerrena unicolor on the formation of a fibrin clot was investigated in the context of potential use as fibrin glue and sealant enhancers and potential wound healing agents. With the use of ROTEM thromboelastometry, we demonstrated that, in the presence of fibrinogen and thrombin, the S6 fraction accelerated the formation of a fibrin clot, had a positive effect on its elasticity modulus, and enhanced the degree of fibrin cross-linking. The S5 fraction alone showed no influence on the fibrin coagulation process; however, in the presence of fibrin, it exhibited a decrease in anti-proliferative properties against the HT-29 line, while it increased the proliferation of cells in general at a concentration of 100 µg/mL. Both fractions retained their proapoptotic properties to a lesser degree. In combination with the S6 fraction in the ratio of 1:1 and 1:3, the fractions contributed to increased inhibition of the activity of matrix metalloproteinases (MMPs). This may suggest anti-metastatic activity of the combined fractions. In conclusion, the potential of the fractions isolated from the C. unicolor secretome to be used as a means of improving the wound healing process was presented. The potential for delivering agents with cytostatic properties introduced far from the site of action or exerting a pro-proliferative effect at the wound site with the aid of a fibrin sealant was demonstrated.

Antitumor potential of new low molecular weight antioxidative preparations from the white rot fungus Cerrena unicolor against human colon cancer cells.

The aim of this study was to investigate the anticancer and antioxidant activities of low molecular weight subfractions isolated from secondary metabolites produced by the wood degrading fungus Cerrena unicolor. Human colon cancer cells (stage I) HT-29 and human normal colon epithelial cells CCD 841 CoTr were used in the research. The present study demonstrated that the low molecular weight subfractions exhibited inhibitory activity towards human colon cancer cells HT-29 at a concentration range of 25-200 µg/mL. All 6 subfractions inhibited proliferation of cells down to 47.5-9.2% at the highest concentrations in a dose-dependent manner. The most desired activity was exhibited by subfractions S, 3, 4, and 5, as the proliferation of HT-29 cells was inhibited to the greatest extent (16.5, 47.5, 42.7, and 26.1% of the control, respectively), while the effect on CCD 841 CoTr cells was the mildest (inhibition to 54.4, 71.4, 79.4, and 53.4%, compared to the control, respectively). The microscopic observation revealed that all extracts induced programmed cell death, i.e. apoptosis (up to 44.4% (subfraction 6) towards HT-29 and less than 20% (most fractions) towards CCD 841 CoTr), with no or a significantly low level of necrosis in both cell lines at the same time.

Medial patellofemoral ligament reconstruction in children: A comparative randomized short-term study of fascia lata allograft and gracilis tendon autograft reconstruction.

BACKGROUND: Many surgical procedures have been described to treat recurrent patellar dislocation, but none of these techniques has been successful in all patients. The goal of the study was to evaluate the results of medial patellofemoral ligament reconstruction in children. Two operative procedures were evaluated; a fascia lata allograft and an autologous gracilis graft. METHODS: Forty-four children (27 girls and 17 boys) between 13 and 17 years of age with unilateral recurrent patellar dislocation underwent medial patellofemoral ligament (MPFL) reconstruction. Patients were operated in two orthopedic centers. The 1st group contained 22 patients and surgery was performed using a fascia lata allograft. In the 2nd group of patients which also contained 22 children and autologous gracilis graft was used. The mean age of the patients was 14.9 years and the mean follow-up was 24 months. Preoperatively, all patients were evaluated clinically (Kujala score questionnaire) and radiologically. The same evaluation was used 18 to 30 months postoperatively to estimate the results of our treatment. RESULTS: In 1st group of children operated with cadaver allografts, the Kujala score significantly improved from 73.91 points preoperatively to 94.50 points postoperatively (P < .001). The average duration of operating procedure was 1 hour and 35 minutes. As shown by subjective symptoms, the results in 95% of patients were rated as good or very good. All children returned to full activity. Similar results were obtained in patients in 2nd group, where MPFL was reconstructed with ipsilateral gracilis tendon. Kujala score increased from 70.77 points preoperatively to 94.32 postoperatively (P < .001). Our results were estimated as good or very good in 93% of patients. All patients that were operated returned to full activity. However, median duration of operation was longer and lasted 1 hour and 55 minutes. CONCLUSIONS: Both techniques were effective in the short-term (18-30 months) in treatment of recurrent patellar dislocation. The use of cadaver allograft spares the hamstring muscles and reduces the time of surgery. Therefore, such study appears to be useful because it provides valuable information that would help to guide treatment of this condition in children. Level of evidence II-2.

Cat Scratch Disease in a 1.5-year-old girl - Case report.

INTRODUCTION: The paper is a case report presenting Cat Scratch Disease (CSD) in a 1.5-year- old girl. Bartoneloses, including CSD, are a group of infectious diseases which are rarely detected, therefore there are no statistical data concerning the aetiology, and the incidence of CSD noted in Poland is low in comparison with other European countries. OBJECTIVE: The purpose of the paper is to discuss several problems related to CSD. MATERIAL AND METHODS: A 1.5-year-old girl who was seen in hospital for the sparing use of her left arm when crawling. X-rays showed osteolytic lesions which radiologists described as multi-ocular cyst or infection. As neither clinical examination nor laboratory investigations found pathological signs, the patient was followed-up on an ambulant basis. Repeated x-ray taken 4 weeks later showed increased periosteal proliferation accompanied by pain. The baby was admitted to the Clinic but additional investigations found no pathologies. The baby was consulted by a rheumatologist and haematologist; however, they did not facilitate a definitive diagnosis. As the baby developed, because of a thickening of the soft tissues on the dorsal side of the distal epiphisis in the forearm the doctors decided to inspect the condition operatively. Macroscopic examination found brownish granulated tissue. Suction drainage was inserted and a tissue sample was tested for aerobic and anaerobic bacteria, tuberculosis and borelliosis. The test results were negative. The baby was in good condition, was not pyrexial and suffered from less pain. The diagnostics was further expanded and the baby tested for yersinia, chlamydia, tuberculosis and bartonella, i.e. CSD. The postoperative wound healed soon and radiological bony lesions began to resolve. After a month, we received a positive bartonella test result, the baby tested positively for Bartonella henselae IgG class, which confirmed past or active infection of CSD. A repeated test for B. henselae taken 6 months later showed a lower level of antibodies. CONCLUSIONS: It should be remembered that CSD, which is an extremely rare infection, can be diagnosed despite mediocre clinical and radiological manifestations. Thus, in the case of infections of unexplained aetiology and mediocre manifestations diagnostics should include testing for Bartonella henselae.

Anticancer, antioxidant, and antibacterial activities of low molecular weight bioactive subfractions isolated from cultures of wood degrading fungus Cerrena unicolor.

The aim of this study is to investigate in vitro the anticancer, antioxidant, and antibacterial activities of three low molecular weight subfractions I, II and III isolated from secondary metabolites produced by the wood degrading fungus Cerrena unicolor. The present study demonstrated that the low molecular weight subfractions III exhibited the strongest inhibitory activity towards breast carcinoma cells MDA-MB-231, prostatic carcinoma cells PC3, and breast cancer cells MCF7 with the half-maximal inhibitory concentration (IC50) value of 52,25 µg/mL, 60,66 µg/mL, and 54,92 µg/mL, respectively. The highest percentage of inhibition was noted at a concentration of 300 µg/mL in all the examined tumor lines. A significant percentage (59.08%) of ex-LMSIII inhibition of the MDA-MB-231 tumor line was reached at a concentration of 15 µg/ml, while the concentration applied did not affect normal human fibroblast cells. The low molecular weight subfraction III was the most effective and additionally showed the highest free radical 1,1-diphenyl-2-picryl-hydrazyl scavenging activity (IC50 20.39 µg/mL) followed by the low molecular weight subfraction I (IC50 64.14 µg/mL) and II (IC50 49.22 µg/mL). The antibacterial activity of the tested preparations was evaluated against three microorganisms: Bacillus subtilis, Staphylococcus aureus, and Escherichia coli. The MIC minimal inhibitory concentration (MIC) values for the low molecular weight subfraction I, II, and III showed a stronger inhibition effect on S. aureus than on B. subtilis and E. coli cells. The MIC values for the low molecular weight subfraction II against S. aureus, B. subtilis, and E. coli were 6.25, 12.5, and 100 mg/mL, respectively.

Biomechanical parameters of the BP-enriched bone cement.

Bisphosphonates (BPs) are well-known substances with very efficient antiresorptive properties. Their beneficial actions are useful not only in achieving better bone mineral density but also in improving bone microarchitecture, strength and, consequently, its quality. Surgical cement, being a polymer composite, is required to be highly biocompatible and biotolerant. The goal of the presented study was to assess whether the enrichment of cement with pamidronate has changed its biomechanical properties. We compared the biomechanical parameters of clean bone cement and BP-enriched bone cement, which were both used formerly in our rat models. Biomechanical properties of BP-enriched bone cement are defined by two basic terms: stress and strain, which are caused by the influence of external force. In the investigatory process of the bone's biomechanical parameters, the compressive test and the three-point flexural tests were used. During the three-point flexural investigation, the sample was supported at both ends and loaded in the middle, resulting in a flexure. After a specific range of flexure, the sample was fractured. In obtained results, there were no significant differences in the values of the stress determined at the point of maximal load and the energy stored in the samples for proportional stress-strain limit (elastic region). There were also no significant differences in the density of the samples. The study shows that the enrichment of bisphosphonates causes yielding of the bone cement material. In the presented data, we conclude that use of pamidronate implanted in bone cement did not have a detrimental effect on its biomechanical properties. Therefore, the obtained results encouraged us to perform further in vivo experiments which assess the biomechanical properties of bones implanted with BP-enriched bone cement.

Evaluation and management of pulseless pink/pale hand syndrome coexisting with supracondylar fractures of the humerus in children.

Elbow region fractures are the most common injuries in children. Among them, supracondylar fractures of the humerus are the most frequent. Massive displacement of the fractured bone causes severe injury to the soft tissue of that particular region. As a result, various types of injuries to the brachial artery such as entrapment, laceration, spasm of the vessel, and the presence of an intimal tear or thrombus formation are usually observed. The main goal of this study was to present our approach to children with supracondylar humerus fractures associated with brachial artery injuries. We would especially like to emphasize the necessity for other conservative or operative treatment concerning pulseless hand symptoms coexisting with supracondylar fractures of the humeral bone in children population. Data from 67 children were evaluated in our study. Supracondylar fractures were classified according to the Gartland's scale. All patients had displaced extension type III injuries. During our follow-up study, we used Flynn's grading system to evaluate functions of the elbow joint, forearm and wrist. Mean follow-up was 18 months; range, 13 months to 4 years. In the follow-up study, very good or good results were achieved in all 32 patients treated conservatively together with 6 patients with pulseless pink hand symptom. Very good or good results were achieved in 88% of 35 patients operated on. Children who, after satisfactory closed reduction, have a well-perfused hand but absent radial pulse do not necessarily require routine exploration of the brachial artery. Conservative treatment should be applied unless additional signs of vascular compromise appear. Thus, exploration of the cubital fossa should be performed only if circulation is not restored by closed reduction.

Retrospective epidemiological study of supracondylar fractures of the humeral bone in children from urban and rural areas of the Lublin region in eastern Poland.

INTRODUCTION: Supracondylar fractures of the humeral bone are frequent injuries in children. It has been affirmed that supracondylar fractures have an excellent prognosis when proper treatment is applied. OBJECTIVE: Present of the statistical relationships between fractures occurring and patient's development period; the relation between development period and site of the fracture and statistical relationship between development period and gender of the patients. Also indicated are the place of residence of the hospitalized patients and time of admission to the Clinic after injury. MATERIALS AND METHOD: Research was based on the data of paediatric patients treated in the Clinic for Paediatric Surgery, Traumatology and Paaediatric Orthopaedics, and Rehabilitation Clinic of the Medical University in Lublin, Poland, between 1986- 2010. An independent Chi-square Test was used for statistical analysis (χ²). RESULTS: The majority of patients were admitted to the Clinic on the day of injury. Of these patients, 71% lived in the urban area of the Lublin region where all the children received medical care in hospital directly after trauma; 29% of children came from the rural areas of the Lublin region, and 10% of them were admitted to hospital 24 or more hours after the injury. CONCLUSION: 71% of patients lived in the urban areas of the Lublin region and the main cause of injury was a fall from a higher level onto an outstretched upper left limb. Most supracondylar fractures of the humeral bone concerned children at school and adolescent age. Despite the fact that some of the hospitalised children lived in the rural areas of the Lublin region, the majority were admitted to the Clinic directly after trauma and received timely treatment.

Effect of implanted bisphosphonate-enriched cement on the trabecular microarchitecture of bone in a rat model using micro-computed tomography.

PURPOSE: Bisphosphonates (BPs) are antiresorptive drugs typically used to inhibit bone resorption. The latest reports show that BPs play an important role in not only achieving better bone mineral density but also in improving bone microarchitecture. The mechanism of action of the BPs is complex and multifactorial. We tried to determine whether there are any changes in the microarchitectural bone structure during local use of BP (Pamifos 60). The aim of this study was to see if BP-enriched cement used in rat models had positive effects on bone formation. METHODS: Research was performed on 40 adult male Wistar rats that were divided into four groups: two control groups and two experimental groups. Rats in the experimental groups were implanted with BP-enriched cement into the bone, while the control group rats were implanted with clean bone cement (without BP). Micro-computed tomography was applied for the investigation of trabecular microarchitecture of the proximal physis of the tibial bone in all animals three and six weeks after surgery. In all microCT images variables such as bone volume density (BV/TV), trabecular thickness (TbTh), trabecular separation (TbSp) and trabecular number (TbN) were used to describe trabecular bone morphometry. RESULTS: The major finding of this study is that using BP-enriched cement results in distinct changes in bone microarchitecture. We showed that local use of pamidronate (Pamifos 60) in orthopaedic cement had a positive effect on bone formation. It significantly changed three variables. We noticed increasing bone volume fraction and trabecular thickness together with decreasing trabecular separation. CONCLUSION: In this paper we demonstrate the efficacy of using BP-enriched cement in vitro in the tibiae of rats. Our most significant finding based on micro-CT picture analysis allows us to start further work on more suitable applications of BP-enriched cement in humans. We believe that future successful experiments will facilitate potential use of BP-enriched cement in clinical applications.

Implanted bisphosphonates in bone cement affect bone markers in rat serum.

PURPOSE: Bisphosphonates (BPs) are antiresorptive drugs that provide important effects on bone turnover. The key to the high efficiency of BPs is their affinity for bone tissue, and their chemical structure provides their molecular mechanism of action. BPs are widely used to treat a variety of diseases that cause excess bone resorption, such as bone metastasis, hypercalcaemia due to malignancy and Paget's disease. The goal of this study was to assess whether the bisphosphonate (Pamifos®) present in bone cement has any effect on bone turnover. In this paper, we present changes in cytokine levels in the serum of rats treated surgically. METHODS: Research was performed on 40 adult male Wistar rats. The rats were divided into four groups: two control groups (A, B) and two experimental groups (C, D). Bone in rats in the experimental groups was implanted with BP-enriched cement, whereas bone in control-groups rats was implanted with clean cement (without BPs). RESULTS: We found a higher concentration of tumour necrosis factor alpha (TNF-α) three weeks after surgery in rats implanted with BP-enriched cement compared with rats implanted with clean cement. After six weeks of treatment, TNF-α levels decreased significantly in rats treated with BP-enriched cement, whereas the control group experienced an increase in TNF-α. The concentration of osteoprotegerin ligand (OPG) was higher in rats with BP implants. We found high levels of receptor activator of nuclear factor kappa-B ligand (RANKL) in rats after implantation of cement without BP in both groups. CONCLUSIONS: We conclude that use of bisphosphonate (Pamifos®), which is present in bone cement, has an effect on bone turnover in that BPs stimulate an increase in OPG and a decrease in RANKL in the bone microenvironment and thus may be an important component of mechanisms that reduce bone resorption. Therefore, the use of BP-enriched cement implants appears to be justified.

Early results of treatment for congenital clubfoot using the Ponseti method.

The purpose of this study was to evaluate the early results of the Ponseti method in reducing extensive corrective surgery rates for congenital idiopathic clubfoot in patients treated in Children's Orthopaedic Clinic and Rehabilitation Department Medical University of Lublin between the years 2007-2011. Thirty-five patients with 47 idiopathic clubfeet were followed prospectively while being managed with the Ponseti method. Clubfoot severity was graded with use of the Dimeglio system. The initial correction was achieved, and early results were measured by using Pirani scoring method.

Florid-reactive periostitis of the phalanges: a 2-case report.

Florid-reactive periostitis, also known as fibroosseous pseudotumor, is a benign bone lesion. It occurs rarely among pediatric patients. Usually, it affects the phalanges of the hands and feet, where most lesions concern the proximal phalanx. Girls are more commonly affected than boys. The lesion appears benign radiographically but on histologic examination could be mistaken for a variety of malignant lesions, especially osteosarcoma. Therefore, histologic evaluation is a must for distinguishing this benign lesion from malignant and infectious etiologies. We present 2 patients with florid periactive periostitis of the phalanges who were treated in our clinic. After physical and microscopy examination of the soft and bone tissues, we found no evidence of malignant process. Because of a definitive diagnosis and the benign nature of the histologic findings, a conservative treatment, that is, no surgical intervention, with a close supervision of our patients was used. During follow-up observations in both patients, we did not notice any limitations of finger movements or an enlargement of the tumor. These findings confirmed that for cases with no aggressive growth, a conservative treatment may be effective and beneficial for a patient.

Bladder squamous metaplasia of the urothelium - introductory report.

Seven female patients hospitalized in the Surgery Ward of Warsaw's Hospital for Children are described. Nonspecific abdominal pains, recurrent urinary tract infections, and voiding disorders (difficulty in voiding commencement) were the primary causes of hospitalization. Urodynamics and cystoscopy with a histopathology section of the urothelium achieved the diagnosis of nonkeratizing squamous cell metaplasia of the urothelium. The topic is not new, although the diagnosis is particularly rare in the pediatric patient. Until now, the existing world literature has described only a few cases of this kind of metaplasia in children. These findings may be helpful in differential diagnosis of urinary bladder disease. It is postulated that nonkeratizing squamous cell metaplasia of the urothelium, in the future, may lead to malignant changes in the urothelium. Therefore, the therapeutic process merits close monitoring.