Inflamed Wounds Research Articles

A Convolutional Neural Network- Based Deep Learning To Detect Reticulocytes From Human Peripheral Blood

Machine learning approaches are rapidly augmenting, and in some cases, replacing the conventional methods in biomedical data analysis; to reduce time, cost, biases, and the need for sophisticated analytical platforms. Hence, significant interest has been compounded in the integration of automated image analysis for various clinical applications, such as the detection of infected or inflamed wounds, bone fractures or for the purpose of disease diagnosis – such as Plasmodium parasites or circulating tumour cells in blood. Here, we report the development of a Convolutional Neural Network (CNN)-based method on CPU to distinguish and count immature human red blood cells known as reticulocytes from blood smears. Reticulocytes represent a heterogeneous and relatively small percentage of cells in peripheral blood, and contain residual RNA in complex with proteins which generates thread-like patterns when stained with New Methylene Blue (NMB) dye. We used more than 200 NMB-stained images from leukocyte-depleted blood to train and optimize the model for immature reticulocytes (stained positive with NMB, intensity and pattern of which depends on the developmental stage of the reticulocyte) and mature RBCs (no staining with NMB). The training performance evaluation metrics demonstrated a mean average precision (mAP50) of 0.88, a precision of 0.83, a recall of 0.88, and an F1 score of 0.87. Our model was able to successfully count reticulocytes with accuracy more than 90% from unknown samples which were subsequently cross-verified through microscopy and counting. Given the importance of reticulocyte dynamics in blood and its clinical relevance, the newly developed model will find important easy to adopt biomedical applications that can be achieved on a simple PC.

A case report of iliopsoas abscess and literature review.

Iliopsoas abscess is a rare acute medical condition. It usually occurs because of the spread of infection from adjacent structures and hematogenous spread. Clinical features include fever, backache, radiating nerve root pain, and leg weakness. When sepsis occurs, prompt recognition is required to initiate appropriate antimicrobial therapy and surgical drainage. A 65-year-old male presented to the outpatient department with a 2-day history of lower back, hip, and leg pain, for which analgesics were administered. During hospitalization, he experienced deterioration, becoming febrile, hypoxic, hypotensive, tachycardiac, and delirious. The patient was then intubated and ventilated. His family reported an additional history of acupuncture for back pain, which sustained an inflamed wound on his right forearm. Abdominal computed tomography was performed, which confirmed bilateral iliopsoas abscess without involvement of intra-abdominal organs. A preliminary report of blood culture revealed Gram-positive cocci. Echocardiography showed vegetation on the aortic valve, and moderate aortic regurgitation was sustained. He was started on vancomycin along with piperacillin-tazobactam. Ultrasound-guided percutaneous drainage was inserted into the bilateral abscess. Pus and blood yielded methicillin-sensitive Staphylococcus aureus. He remained septic. The repeat computed tomography showed the right abscess enlarged. A repeated echocardiogram showed that the vegetation increased. Further incision and surgical drainage were performed with continuous wash-out. His condition improved after management and he was discharged to a regional hospital for ongoing care. Prompt diagnosis and surgical treatment are essential to improve patient outcomes. The unique aspect of this case is the persistence of the methicillin-sensitive Staphylococcus aureus infection. Centralized surgical services are pivotal in conjunction with robust antimicrobial regimens. This case reinforces the importance of high clinical suspicion of an unknown source of sepsis.

Exploring Microemulsion Systems for the Incorporation of Glucocorticoids into Bacterial Cellulose: A Novel Approach for Anti-Inflammatory Wound Dressings.

The effective pharmacological treatment of inflamed wounds such as pyoderma gangraenosum remains challenging, as the systemic application of suitable drugs such as glucocorticoids is compromised by severe side effects and the inherent difficulties of wounds as drug targets. Furthermore, conventional semi-solid formulations are not suitable for direct application to open wounds. Thus, the treatment of inflamed wounds could considerably benefit from the development of active wound dressings for the topical administration of anti-inflammatory drugs. Although bacterial cellulose appears to be an ideal candidate for this purpose due to its known suitability for advanced wound care and as a drug delivery system, the incorporation of poorly water-soluble compounds into the hydrophilic material still poses a problem. The use of microemulsions could solve that open issue. The present study therefore explores their use as a novel approach to incorporate poorly water-soluble glucocorticoids into bacterial cellulose. Five microemulsion formulations were loaded with hydrocortisone or dexamethasone and characterized in detail, demonstrating their regular microstructure, biocompatibility and shelf-life stability. Bacterial cellulose was successfully loaded with the formulations as confirmed by transmission electron microscopy and surprisingly showed homogenous incorporation, even of w/o type microemulsions. High and controllable drug permeation through Strat-M® membranes was observed, and the anti-inflammatory activity for permeated glucocorticoids was confirmed in vitro. This study presents a novel approach for the development of anti-inflammatory wound dressings using bacterial cellulose in combination with microemulsions.

In English

The use of wound dressings is gaining more and more popularity, especially in the field of tactical and military medicine. Developing wound dressings capable of facilitating wound treatment and reducing healing time is one of the challenges of modern science. So, sodium alginate (Alg) is a good candidate for the development of wound dressings due to its bio- and hemocompatibility and biodegradability. However, Alg has its shortcomings, which can be dispatched by modification. The purpose of this work was to investigate the effect of Alg modification on the kinetics of ethonium release from crosslinked with Ca2+ ions samples. For this purpose, a method of Alg modifying with octane-1-amine was developed without the use of organic solvents and with the use of 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDCl) as an initiator. The optimal parameters of alginate modification process were defined as 60 °С temperature and 24 hours duration. Physicochemical methods confirmed the success of the modification. Films based on the alginate modified with octane-1-amine (AlgM) were obtained using a calcium chloride solution as a crosslinker. The kinetics of swelling was studied and we found that the degree of swelling of the sample based on AlgM after 10 minutes is twice as large (α = 0.71) as for Alg (α = 0.37), which indicates a faster release of drugs. It has been found that the kinetics of release of ethonium depends not only on the kinetics of swelling but also on the chemical nature of the drug. The ethonium was immobilised in alginate films as a model of bactericidal drug. The kinetics of ethonium release was studied at different pH values corresponding to the pH of healthy skin (5.5), open wounds (7.2) and inflamed wounds (8.2). It was found that the release of ethonium from the sample based on AlgM is more pH-sensitive and prolonged, compared to the sample based on Alg. This effect is explained by the appearance of an additional mechanism of retention of ethonium by AlgM due to hydrophobic-hydrophobic interactions in the films. The prolonged release properties observed in the drug-loaded samples make them promising candidates for the development of targeted drug delivery systems and wound dressings, which are particularly relevant for the treatment of chronic and burn wounds. Future research will focus on optimizing the crosslinking method and exploring potential applications of modified alginate-based materials in biomedical sciences.

Is my wound infected? A study on the use of hyperspectral imaging to assess wound infection.

Clinical signs and symptoms (CSS) of infection are a standard part of wound care, yet they can have low specificity and sensitivity, which can further vary due to clinician knowledge, experience, and education. Wound photography is becoming more widely adopted to support wound care. Thermography has been studied in the medical literature to assess signs of perfusion and inflammation for decades. Bacterial fluorescence has recently emerged as a valuable tool to detect a high bacterial load within wounds. Combining these modalities offers a potential objective screening tool for wound infection. A multi-center prospective study of 66 outpatient wound care patients used hyperspectral imaging to collect visible light, thermography, and bacterial fluorescence images. Wounds were assessed and screened using the International Wound Infection Institute (IWII) checklist for CSS of infection. Principal component analysis was performed on the images to identify wounds presenting as infected, inflamed, or non-infected. The model could accurately predict all three wound classes (infected, inflamed, and non-infected) with an accuracy of 74%. They performed best on infected wounds (100% sensitivity and 91% specificity) compared to non-inflamed (sensitivity 94%, specificity 70%) and inflamed wounds (85% sensitivity, 77% specificity). Combining multiple imaging modalities enables the application of models to improve wound assessment. Infection detection by CSS is vulnerable to subjective interpretation and variability based on clinicians' education and skills. Enabling clinicians to use point-of-care hyperspectral imaging may allow earlier infection detection and intervention, possibly preventing delays in wound healing and minimizing adverse events.

The bactericidal effects of 632.8 nm He-Ne laser on Staphylococcus aureus colonies

The bactericidal effect of 632.8 nm low level laser has been studied in order to point out both the effective power and laser exposure time on Staphylococcus aureus, which is reported to be involved in several dermatology problems. Low level laser has been reported to be useful for infected wounds, tissue necrosis, nerve injury, osteoarthritis or other chronic pain syndromes. Numerous studies have been conducted to quantify the effective laser parameters, i.e. dose, power, and exposure time, which ultimately leads toward clinical implementation. Staphylococcus aureus bacteria colonies were exposed to laser doses with powers of both 1 and 3 mW at different exposure times varies between 3 to 30 minutes. The bacterial colonies were isolated from a patient with inflamed wounds. Two sets of bacterial colonies were prepared to be exposed to laser beam. Next, the bacterial colonies were compared before and after exposing them to laser doses. The results showed that laser sessions have reduced the number of the bacterial colonies for both doses; 1 and 3 mw at the different exposure times and concentrations. The results revealed significant dose dependent bactericidal effects of He-Ne laser on Staphylococcus aureus at 3 mW for 30 minutes, which was found to be more effective in reducing the amount of bacteria to the less than 2% of its initial count. The results exhibited the reduction of the number of colonies as a function of exposure time. Appropriate doses of 632.8 nm can kill Staphylococcus aureus, suggesting that a similar effect may be used in clinical cases of bacterial infection.

Is the duration of diabetic foot ulcers an independent risk factor for developing diabetic foot osteomyelitis?

ObjectiveThis study aimed to determine whether the prolonged duration of diabetic foot ulcers was associated with an increased incidence of diabetic foot osteomyelitis. Study designA retrospective cohort study MethodsThe medical records of all patients who participated in the diabetic foot clinic between January 2015 and December 2020 were reviewed. Patients with new diabetic foot ulcers were monitored for diabetic foot osteomyelitis. The collected data included the patient’s profile, comorbidities and complications, the ulcer profile (area, depth, location, duration, number of ulcers, inflammation, and history of the previous ulcer), and outcome. Univariate and multivariate Poisson regression analyses were used to assess risk variables for diabetic foot osteomyelitis. ResultsEight hundred and fifty-five patients were enrolled; 78 developed diabetic foot ulcers (cumulative incidence 9% over 6 years, average annual incidence 1.5%) and among these diabetic foot ulcers, 24 developed diabetic foot osteomyelitis (cumulative incidence 30% over 6 years, average annual incidence of 5%, incidence rate 0.1/person-year). Statistically significant risk factors for the development of diabetic foot osteomyelitis were ulcers that were deep to the bone (adjusted risk ratio 2.50, p = 0.04) and inflamed wounds (adjusted risk ratio 6.20, p = 0.02). The duration of diabetic foot ulcers was not associated with diabetic foot osteomyelitis (adjusted risk ratio 1.00, p = 0.98). ConclusionThe duration was not an associated risk factor for diabetic foot osteomyelitis, while bone-deep ulcers and inflamed ulcers were found to be significant risk factors for the development of diabetic foot osteomyelitis.

Use of wound edge inversion (epibole) to generate recalcitrant and inflamed diabetic wounds.

Robust and predictive pre-clinical models of recalcitrant diabetic wounds are critical for advancing research efforts toward improving healing. Murine models have logistic and genetic benefits versus larger animals; however, native murine healing inadequately represents clinically recalcitrant wounds in humans. Furthermore, current humanization techniques employing devices, deleterious mutations or chemical agents each carry model-specific limitations. To better replicate human wounds in a mouse, we developed a novel wound-edge inversion (WEI) technique that mimics the architecture of epibole and mitigates contracture, epithelialization, and consequently wound closure. In this study, we evaluated the reliability and durability of the WEI model in wild-type and obese diabetic mice and compared to healing after (i) punch biopsy, (ii) mechanical/silicone stenting or (iii) exogenous oxidative stressors. In wild-type mice, WEI demonstrated favourable closure characteristics compared to both control and stented wounds, however, wounds progressed to closure by 4 weeks. In contrast, diabetic WEI wounds persisted for 6-10 weeks with reduced contracture and epithelialization. In both diabetic and wild-type mice, WEI sites demonstrated persistence of inflammatory populations, absence of epithelialization, and histologic presence of alpha-SMA positive granulation tissue when compared to controls. We conclude that the WEI technique is particularly valuable for modelling recalcitrant diabetic wounds with sustained inflammation and dysfunctional healing.

An all-in-one CO gas therapy-based hydrogel dressing with sustained insulin release, anti-oxidative stress, antibacterial, and anti-inflammatory capabilities for infected diabetic wounds

To effectively treat diabetic wounds, the development of versatile medical dressings that can long-term regulate blood glucose and highly effective anti-oxidative stress, antibacterial and anti-inflammatory are critical. Here, an all-in-one CO gas-therapy-based versatile hydrogel dressing (ICOQF) was developed via the dynamic Schiff base reaction between the amino groups on quaternized chitosan (QCS) and the aldehyde groups on benzaldehyde-terminated F108 (F108-CHO) micelles. CORM-401 (an oxidant-sensitive CO-releasing molecules) was encapsulated in the hydrophobic core of F108-CHO micelles and insulin was loaded in the three-dimensional network structure of ICOQF. The dynamic Schiff base bonds not only endowed ICOQF with good tissue adhesion, injectability and self-healing, but also gave it sustained and controllable insulin release ability. In addition, ICOQF could quickly generate CO in inflamed wound tissue by consuming reactive oxygen species. The generated CO could effectively anti-oxidative stress by activating the expression of heme oxygenase; antibacterial by inducing the rupture of bacterial cell membranes and mitochondrial dysfunction and inhibiting the synthesis of adenosine triphosphate; and anti-inflammatory by inhibiting the proliferation of activated macrophages and promoting the polarization of the M1 phenotype to the M2 phenotype. Due to these outstanding properties, ICOQF significantly promoted the healing of STZ-induced MRSA-infected diabetic wounds accompanied by good biocompatibility. This study clearly shows that ICOQF is a versatile hydrogel dressing with great application potential for the management of diabetic wounds. Statement of significanceThe development of some versatile hydrogel dressings that can not only provide a prolonged and controlled insulin release property but also utilize a non-antibiotic treatment modality for highly effective antibacterial, anti-inflammatory, and anti-oxidative stress effects is vital for the successful treatment of diabetic wounds. Herein, we developed an all-in-one CO gas-therapy-based versatile hydrogel dressing (ICOQF) with sustained and controllable insulin release abilities. Moreover, ICOQF could not only quickly release CO in the inflamed wound tissue by consumption of reactive oxygen species but also utilize the generated CO to highly effectively anti-oxidative stress, antibacterial, and anti-inflammatory. ICOQF therapy substantially promoted the healing of STZ-induced MRSA-infected diabetic wounds. Overall, this work provides a multifunctional hydrogel dressing for the management of diabetic wounds.

Chronic wounds healing by innovative herbal medicine and Buddhist ways to prevent disability in diabetic patients

The objectives of this research were 1) to study the properties of both scientific and traditional medicinal herbs to produce innovative herbal medicine for chronic wounds healing in diabetic patients, and 2) to analyze chronic wounds healing by innovative herbal medicine and Buddhist ways to prevent disability in 30 diabetic patients in Mahasarakham, Thailand. The results revealed that 19 medicinal herbs have properties in managing the blood, vascular, lymphatic system, the main cause of chronic wounds, reducing blood viscosity, purifying blood vessels, flushing out waste water, and healing wounds. 5 medicinal herbs were used to produce innovative herbal medicine, and Buddhist principles were used in research and for informants. Chronic wounds in diabetic patients have three characteristics: peripheral neuropathy, ischemic wounds, and infectious wounds. Herbal Oil Formula 1 and Herbal Infusion Formula 2 were used as basic medicines for healing, Herbal Capsule Formula 3 for inflamed wounds, Herbal Infusion for wounds with dark secretions and rotten blood. Wounds with a lot of secretions, foul odors, and full wounds were healed by Mangosteen Peel Powder Formula 5. Accordingly, 100% of all 3 types of chronic wounds were healed. This saved all patients from disability.

Control of Staphylococcal-mediated endogenous protease activity alters wound closure time in a complex wound model

BackgroundElevated protease activity is a characteristic feature of chronic wounds, where the inflammatory phase of wound healing is prolonged. The choice of dressings in treatment of chronic wounds can change the nature of the wound base and have a significant impact on healing. ObjectiveTo evaluate the impact of oxidised regenerated cellulose/collagen dressings on Staphylococcal-mediated protease activity in an inflamed wound model. MethodsWe developed an in vitro 3D inflamed wound model, and simulated inflammation by exposing the models to Staphylococcal spent culture supernatant. Protease activity and wound healing were assessed in the presence/absence of the dressings. ResultsHistological analysis of the wound model revealed two distinct layers, an epidermal and dermal layer, similar to the organisation of human skin. Inflammation with Staphylococcal spent culture supernatant elevated protease levels by 1.7x and consequently prevented the wound from progressing to the proliferative phase of healing, without having a negative effect on cell viability. Adding a collagen dressing, known to have non-specific protease modulating properties, reduced Staphylococcal-mediated protease activity back to baseline, with a concomitant reduction in wound closure time. Inflamed wounds thus resembled unwounded skin after 10 days of treatment with the dressings. ConclusionOur findings support the further evaluation and use of oxidised regenerated cellulose/collagen dressings for inflamed, non-healing wounds in the clinical setting. The model used in this study has the potential to be applied in preclinical research; to test wound dressing performance, such as healing and cell viability, and to also assess key markers of inflammation.

Diversity and prevalence of ticks in the goats in lowland Nepal.

Ticks are very harmful blood sucking protozoans and rickettsial diseases in livestock and cause an adverse economy of the world, including tropical regions like Nepal. Thus, the current study was conducted to determine the distribution, prevalence, and the effects of ticks in the goat of the Rapti Municipality, Chitwan, Nepal. Ticks were picked from different body parts of goats (n=473) and identified to species level by using morphological identification keys under a microscope. The study revealed that 161 (34%) of goats were infested and females had highest prevalence of ticks compared to males (41% vs 28%). A total of 14 species of ticks of genera Haemaphysalis, Amblyomma, Rhipicephalus, Ixodes and Dermacentor were identified. All tick-positive goats were suffered from inflamed wounds with various skin manifestations. In conclusion, this study suggested that appropriate control measures for ticks need to be employed in the study area for economic goat production.

Therapeutic arteriogenesis by factor-decorated fibrin matrices promotes wound healing in diabetic mice

Chronic wounds in type-2 diabetic patients present areas of severe local skin ischemia despite mostly normal blood flow in deeper large arteries. Therefore, restoration of blood perfusion requires the opening of arterial connections from the deep vessels to the superficial skin layer, that is, arteriogenesis. Arteriogenesis is regulated differently from microvascular angiogenesis and is optimally stimulated by high doses of Vascular Endothelial Growth Factor-A (VEGF) together with Platelet-Derived Growth Factor-BB (PDGF-BB). Here we found that fibrin hydrogels decorated with engineered versions of VEGF and PDGF-BB proteins, to ensure protection from degradation and controlled delivery, efficiently accelerated wound closure in diabetic and obese db/db mice, promoting robust microvascular growth and a marked increase in feeding arterioles. Notably, targeting the arteriogenic factors to the intact arterio-venous networks in the dermis around the wound was more effective than the routine treatment of the inflamed wound bed. This approach is readily translatable to a clinical setting.

Antimicrobial Activity of Pulicaria Species from Turkey

Objective: In traditional medicine, the Pulicaria species are used for various disorders such as inflamed wounds, skin diseases, and bronchitis. This study investigated the antibacterial effect of five Pulicaria species in Turkey; (Pulicaria (P) arabica (L.) Cass., P. dysenterica (L.) Bernh., P. odora (L.) Reichb., P. sicula (L.) Moris, P. vulgaris (L.) Gaertn.) against certain significant pathogenic gram-negative and gram-positive reference bacteria. Material and Method: Four extracts (decoction, infusion, aqueous, and ethanol (EtOH) extracts) were prepared from the Pulicaria species. The antimicrobial activity of the samples was examined against reference organisms; Bacillus (B) subtilis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Methicillin-resistant Staphylococcus (S) aureus (MRSA), Proteus mirabilis, S. aureus and Pseudomonas aeruginosa. The minimal bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) ranges of the extract samples were demonstrated based on a microbroth dilution method. Results: The EtOH extracts of the studied four Pulicaria species were found to be weakly active against gram-positive bacteria such as B. subtilis, MRSA and S. aureus but the EtOH extract of P. dysenterica showed exceptionally good activity against the reference strains of S. aureus, MRSA. No antimicrobial activity was detected in the infusion, decoction, and aqueous extracts. Conclusion: The Pulicaria species, especially P. dysenterica could be evaluated as antimicrobial agents. Further studies with the extracts and essential oils from Pulicaria sp. on other bacteria and pathogenic fungi should be performed.

Open tibial fracture with severe soft tissue injury and bone loss managed with ipsilateral fibular transport and its complications: a case report

<p class="abstract">Massive segmental bone defects of tibia present as a challenging task to manage specially when associated with extensive soft tissue injury. A 30 year old male presented to Paras HMRI hospital, Patna, post road traffic accident with Gustilo Anderson 3B comminuted open tibia shaft fracture and with an external fixator in situ with a grossly inflamed and infected wound. Initially patient was managed with serial wound debridement and skin grafting was done early to obtain adequate soft tissue coverage. The patient then underwent application of Ilizarov external fixator with plan of one level fibular osteotomy for ipsilateral fibular transport. With good outcome of the procedure clinically and radiologically, Ilizarov fixator was removed after time duration of about 1.3 years and limb was immobilized in plaster of Paris (POP) cast which was removed after 8 weeks. Within 1 month of removal of POP cast the patient presented to hospital again with complaints of pain and instability when his leg was run over by his child’s bicycle while playing. Diagnosed as fracture of proximal (transported) fibula he was managed then with locking plates; one of which was used as an internal fixator and the other as external fixator which was outside the body and acted as a support to the operated limb. After about 1 year the external locking plate was removed and patient was able to bear weight on his extremities. Despite various modalities to treat massive tibial gap, fibular transport procedure with Ilizarov external fixator seems to be the most viable option.</p>

Chitosan/nanocellulose‐based bionanocomposite films for controlled betamethasone and silver sulfadiazine delivery

AbstractBionanocomposite films based on chitosan and nanocellulose (nanocrystals or nanofibrils) have gained considerable attention for biomedical applications, especially for wound dressings. However, the development of these films as controlled drug release dressings is still under‐exploited. Therefore, this work aimed to design chitosan/nanocellulose‐based bionanocomposite films, loaded by betamethasone or silver sulfadiazine, as functional dressings. The films were obtained by solvent casting and characterized by physicochemical, mechanical, barrier properties, in vitro drug release, and antimicrobial activity. The nanocellulose type, physical state, and content caused influence on the film's properties providing different physical, barrier, and drug release profiles. They are semi‐occlusive and mechanically resistant; the drug release is controlled, and possesses antimicrobial activity. In conclusion, the developed biodegradable bionanocomposite films are promising as active dressings for controlled drug delivery in the wound site and have specific applications according to their features to treat inflamed and purulent wounds, non‐infectious dry wounds, and infectious wounds.

A collagen-AS/εPLL bilayered artificial substitute regulates anti-inflammation and infection for initial inflamed wound healing.

Despite the development of advanced tissue engineering substitutes, inflammation is still a significant problem that can arise from inflamed burn injuries, chronic wounds, or microbial diseases. Although topical wound dressing accelerates healing by minimizing or preventing the consequences of skin inflammation, there remains a need for the development of a novel substitute scaffold that can effectively eliminate immoderate inflammation and infection in the initial phase of the healing meachanism. In this study, an artificial skin substitute scaffold fabricated with asiaticoside (AS) and epsilon-poly-L-lysine (εPLL) was prepared. Upon the release of these bioactive compounds, they accelerate wound healing and inhibit any bacterial infection at the wound site. We determined whether AS and εPLL exhibit anti-inflammatory and bactericidal effects through different mechanisms. Collectively, the collagen-AS/εPLL artificial skin substitute could be a significant therapeutic agent for scar-less rapid wound healing (without infection and inflammation) of initially-inflamed full-thickness wounds.

Efficacy of resveratrol in the wound healing process by reducing oxidative stress and promoting fibroblast cell proliferation and migration.

Oxidative stress (OS) is implicated in the pathogenesis of diabetic, chronic, and inflamed wounds. In these cases, wound healing is difficult and remains a big clinical challenge. As fibroblasts are crucial in wound healing by producing extracellular matrix and wound contraction, the researchers investigated the resveratrol (RES) effects as a potential therapeutic alternative, on an in vitro OS and wound healing model established by 3T3 Swiss Albino fibroblasts. OS was induced by H2 O2 , and 1-100 μM RES doses were applied on cells for up to 48 hours. The cell proliferation was assessed by the population doubling time and bromodeoxyuridine immunocytochemistry. The OS levels were determined by fluorescence staining and all values were calculated by the ImageJ program. The collagen-I expression (COL1A2 mRNA) was evaluated by a real-time PCR. Cell migration was evaluated by wound closure assay and the ultrastructure was evaluated by electron microscopy. The cell proliferation index significantly decreased in H2 O2 incubation and increased with RES application. RES application reduced OS levels and stabilized cell proliferation, but no differences were found in COL1A2 mRNA expression. RES-treated cells showed better proliferation, migration rates, and ultrastructural preservation. RES administration decreases OS levels and improves the healing process by increasing cell proliferation and migration quality, suggesting it is a powerful candidate for the treatment of skin wounds.

Efficient extraction and purification of benzo[c]phenanthridine alkaloids from Macleaya cordata (Willd) R. Br. by combination of ultrahigh pressure extraction and pH-zone-refining counter-current chromatography with anti-breast cancer activity in vitro.

Macleaya cordata (Willd) R. Br. (Papaveraceae family) is a well-known traditional Chinese medicine used to treat muscle pain, inflamed wounds, and bee bites. Benzo[c]phenanthridine alkaloids are the main active ingredients in M. cordata. In this work, sanguinarine and chelerythrine were efficiently extracted and purified by ultrahigh-pressure extraction (UHPE) technique and pH-zone-refining counter-current chromatography (PZRCCC) from M. cordata. To develop an efficient UHPE method followed by an efficient separation technique using PZRCCC for benzo[c]phenanthridine alkaloids from the study plant species, and to evaluate the study samples for anti-breast cancer activity. The optimal extraction conditions were optimised as extraction pressure 200 MPa, extraction solvent 95% ethanol, solid-liquid ratio 1:30 (g/mL) and extraction time 2 min. A two-phase n-hexane/ethyl acetate/i-propanol/water (1:3:1.5:4.5, v/v) solvent system was optimised with 10 mmol triethylamine in the upper phase and 10 mmol trifluoroacetic acid in lower phase in PZRCCC. The sample loading was optimised as 2.50 g. Moreover, the samples were evaluated for anti-breast cancer activity later on. The 2.50 g sample loading yielded 0.45 g of sanguinarine and 0.59 g chelerythrine in one-step separation using PZRCCC. The anti-breast cancer activities of sanguinarine and chelerythrine were found stronger than positive control (vincristine 5.04 μg/mL) with half-maximal inhibitory concentration values of 0.96 and 3.00 μg/mL, respectively. This study showed that the established methods were efficient in extraction (UHPE) and separation (PZRCCC) of the sanguinarine and chelerythrine from M. cordata.

Forensic microbiology reveals that Neisseria animaloris infections in harbour porpoises follow traumatic injuries by grey seals

Neisseria animaloris is considered to be a commensal of the canine and feline oral cavities. It is able to cause systemic infections in animals as well as humans, usually after a biting trauma has occurred. We recovered N. animaloris from chronically inflamed bite wounds on pectoral fins and tailstocks, from lungs and other internal organs of eight harbour porpoises. Gross and histopathological evidence suggest that fatal disseminated N. animaloris infections had occurred due to traumatic injury from grey seals. We therefore conclude that these porpoises survived a grey seal predatory attack, with the bite lesions representing the subsequent portal of entry for bacteria to infect the animals causing abscesses in multiple tissues, and eventually death. We demonstrate that forensic microbiology provides a useful tool for linking a perpetrator to its victim. Moreover, N. animaloris should be added to the list of potential zoonotic bacteria following interactions with seals, as the finding of systemic transfer to the lungs and other tissues of the harbour porpoises may suggest a potential to do likewise in humans.