Photocrosslinked gelatin/chondroitin sulfate/chitosan-based composites with tunable multifunctionality for bone tissue regeneration.

Novel hydrogel-based multifunctional systems prepared utilizing photocrosslinking and freeze-drying processes (PhotoCross/Freeze-dried) dedicated for bone tissue regeneration are presented. Fabricated materials, composed of methacrylated gelatin, chitosan, and chondroitin sulfate, possess interesting features including bioactivity, biocompatibility, as well as antibacterial activity. Importantly, their degradation and swellability might be easily tuned by playing with the biopolymeric content in the photocrosllinked systems. To broaden the potential application and deliver the therapeutic features, mesoporous silica particles functionalized with methacrylate moieties decorated with hydroxyapatite and loaded with the antiosteoporotic drug, alendronate, (MSP-MA-HAp-ALN) were dispersed within the biopolymeric sol and photocrosslinked. It was demonstrated that the obtained composites are characterized by a significantly extended degradation time, ensuring optimal conditions for balancing hybrids removal with the deposition of fresh bone. We have shown that attachment of MSP-MA-HAp-ALN to the polymeric matrix minimizes the initial burst effect and provides a prolonged release of ALN (up to 22 days). Moreover, the biological evaluation in vitro suggested the capability of the resulted systems to promote bone remodeling. Developed materials might potentially serve as scaffolds that after implantation will fill up bone defects of various origin (osteoporosis, tumour resection, accidents) providing the favourable conditions for bone regeneration and supporting the infections' treatment.

Metastasis of tongue cancer locatedunilaterally, not crossing the midline,to the lymph nodes of the neck.

<br><b>Introduction:</b> In recent years, there has been a steady increase in the incidence of tongue cancer. Rapid asymptomatic growth, early metastases, and crossing lymphatic drainage of the tongue reduce the chances of successful therapy. The presence of metastases in cervical lymph nodes is also an important prognostic factor.</br> <br><b>Aim:</b> The aim of the study is a retrospective analysis of the incidence of unilateral and bilateral metastases within the cervical lymphatic system in patients with unilateral tongue cancer that does not cross the midline.</br> <br><b>Materials and methods:</b> A retrospective study of a group of patients hospitalized and treated for malignant tongue cancers at the Clinical Ward of Cranio-Maxillo-Facial Surgery of the Department of Otolaryngology and Laryngological Oncology of the Military Institute of Medicine-National Research Institute in Warsaw in 2012-2023. As many as 106 patients treated for unilateral tongue cancer that has not crossed the midline were selected, who also underwent unilateral or bilateral cervical lymphadenectomy, apart from primary tumor resection and reconstruction. The analysis was based on the medical records of the patients, results of imaging and histopathological examinations, and surgical procedure protocols.</br> <br><b>Results:</b> In a group of 106 patients with unilateral tongue cancer, in whom the cancer did not cross the midline, 47 patients underwent unilateral lymphadenectomy, of which 16 had lymph node metastases. Bilateral lymph node removal was performed in 59 patients. In 26 cases metastases were detected only in ipsilateral lymph nodes, and in 13 cases metastases were found in contralateral lymph nodes. Among 106 patients, occult metastases were detected in 21 patients.</br> <br><b>Conclusions:</b> Due to the crossing of the lymphatic drainage of the tongue, which allows for the possibility of contralateral metastases, and the impact of the operation on oncological efficacy, it is reasonable to perform bilateral cervical lymphadenectomy.</br>.

The influence of the essential oil extracted from hops on monolayers and bilayers imitating plant pathogen bacteria membranes.

Many plant-derived compounds possess antimicrobial, antioxidant and even anticancer activities. Therefore, they are considered as substances that can be used instead of synthetic compounds in various applications. In this work, the essential oil from hop cones was extracted and analyzed, and then its effects on model bacteria membranes were studied to verify whether the hop essential oils could be used as ecological pesticides. The experiments involved surface pressure-area measurements, penetration studies and Brewster angle microscopy (BAM) imaging of lipid monolayers as well as hydrodynamic diameter, zeta potential, steady-state fluorescence anisotropy and Cryo-Transmission Electron Microscopy (cryo-TEM) measurements of liposomes. Finally the bactericidal tests on plant pathogen bacteria Pseudomonas syringae pv. lachrymans PCM 1410 were performed. The obtained results showed that the components of the essential oils from hop cones incorporate into lipid monolayers and bilayers and alter their fluidity. However, the observed effect is determined by the system composition, its condensation and the oil concentration. Interestingly, at a given dose, the effect of the essential oil on membranes was found to stabilize. Moreover, BAM images proved that hop oil prevents the formation of a large fraction of a condensed phase at the interface. Both the studies on model membranes as well as the in vitro tests allow one to conclude that the hop essential oil could likely be considered as the candidate to be used in agriculture as a natural pesticide.

[Differentiation of an integrated and episomal HPV-16 DNA using real-time PCR in cervical specimens of women diagnosed with intraepithelial lesions and invasive cervical cancer]. / Róznicowanie postaci zintegrowanej i episomalnej DNA HPV-16 metoda real-time PCR w wydzielinie szyjki macicy kobiet z rozpoznan sródnablonkowa neoplazja i rakiem szyjki macicy.

UNLABELLED: Persistent high-risk HPV infection, especially HPV-16, is considered to be an important step in the process of cervical carcinogenesis. Integration of viral DNA into the host genome through the destruction of HPV E2 sequences, increases the expression of viral proteins E6 and E7 and their participation in the transformation of cervical cancer. OBJECTIVE: The aim of this study was to apply real-time PCR (RT-PCR) to assess the prevalence of integrated and episomal HPV-16 DNA and determine viral DNA load in women with cervical intraepithelial lesions and invasive cervical cancer MATERIAL AND METHODS: A total of 84 women infected with HPV-16, including 44 with LSIL, 7 with HSIL and 33 with invasive cervical cancer participated in the study Cervical specimens were collected using the cytobrush. The presence of a sequence of E2 and E6 HPV-16 and human gene RNasy P was detected by quantitative RT-PCR. The viral load presented as the form of the virus genome copy numbers per 1,000 cells. RESULTS: The integrated form of HPV-16 genome was found in 97% of women with cervical cancer. In women with LSIL and HSIL mixed form (simultaneous occurrence of an integrated and episomal form) of the viral genome (84% and 57%, respectively) prevailed. The frequency of the integrated HPV-16 DNA increased with progression of dysplastic lesions of the cervix (p<0.001). Statistically significant differences in average number of copies of the virus in women with LSIL and HSIL compared to patients with cancer (p<0.001) were observed. The highest viral load was detected in women demonstrating an integrated HPV-16 DNA. CONCLUSIONS: Quantitative analysis of the sequence of E2 and E6 HPV-16 tested by RT-PCR can be used to determine the degree of integration of the viral genome and quantitative evaluation of viral load in clinical material. It can also serve as an additional parameter defining risk of progression of transformation in the cervix.

Effects of tea tree (Melaleuca alternifolia) oil on Staphylococcus aureus in biofilms and stationary growth phase.

Tea tree oil (TTO) is known for its antimicrobial activity. In this study, we determined whether TTO is effective against Staphylococcus aureus in biofilms and how TTO activity is affected by the S. aureus growth phase. All clinical strains tested were killed by TTO both as planktonic cells and as biofilms. The minimum biofilm eradication concentration was usually two times higher than the minimum bactericidal concentration, yet it was never higher than 1% v/v. The fastest killing of biofilm occurred during the first 15min of contact with TTO and was not influenced by increasing TTO concentration above 1% v/v. Planktonic stationary phase cells exhibited decreased susceptibility to TTO compared with exponential phase cells. The killing rate for stationary phase cells was also less affected by increasing TTO concentration than that for exponential phase cells. These data show that TTO efficiently kills S. aureus in the stationary growth phase and within biofilms and is therefore a promising tool for S. aureus eradication.

SEB-induced T cell apoptosis in atopic patients--correlation to clinical status and skin colonization by Staphylococcus aureus.

INTRODUCTION: We asked whether in atopic dermatitis (AD) increased T cell apoptosis in staphylococcal enterotoxin B (SEB)-activated cultures of peripheral blood mononuclear cells (PBMCs) is characteristic of the exacerbation of the disease or connected with skin colonization by Staphylococcus aureus. MATERIAL/METHODS: The clinical status of the patients was evaluated using the SCORAD index. The number of bacteria colonizing patients' skin lesions was determined by the cfu method. Mononuclear cells isolated from peripheral blood were stimulated by SEB and the apoptosis of CD3+ cells in culture was determined by flow cytometry using the monoclonal antibody APO2.7. The cytokine production in the culture supernatants was determined by ELISA and Cytometric Bead Array kits. RESULTS: T cell apoptosis was increased, while the production of interferon (IFN)-gamma was reduced in cultures of PBMCs of AD patients during exacerbation. The proportion of CD3+ APO2.7+ cells positively correlated with the density of S. aureus recovered from skin lesions, but not with SCORAD index. By contrast, SCORAD index, but not S. aureus density, negatively correlated with IFN- gamma production. Furthermore it was found that the presence of S. aureus on uninvolved skin distinguishes a group of severe cases with high serum IgE level, increased T cell apoptosis, and reduced production of tumor necrosis factor alpha in SEB- -stimulated cultures. CONCLUSIONS: Among AD patients the increased activation-induced T cell apoptosis observed in SEB- -stimulated cultures is related to skin colonization by S. aureus. The presence of bacteria on uninvolved skin is a feature of a distinct group of AD patients.

Antibacterial hemoglobin peptides in human menstrual blood.

This work documents that normal menstrual vaginal blood of healthy females is exceptionally rich in hemocidins--hemoglobin (Hb) fragments having bactericidal properties. The peptide fractions were isolated from the plasma of vaginal discharge of three healthy nulliparous women and subjected to identification by automatic sequencing as well as by mass spectrometry. All 44 identified peptides originate from Hb (mainly from the N-terminal part of alpha-globin) and all demonstrated differential killing activity toward Escherichia coli. The screening of antimicrobial activity was performed using two synthetic peptides identical to those found in menstrual blood. These peptides were active mainly toward Gram-negative bacteria and to a less degree toward Gram-positive bacteria. Our results confirm recent observations that Hb-derived fragments manifest pronounced antibacterial activity and suggest that these peptides help in maintaining human vaginal homeostasis during physiologic menstrual bleeding.