Severe Tissue Necrosis Research Articles

Sodium Hypochlorite Accidents in Endodontic Practice: Clinical Evidence and State of the Art.

Sodium hypochlorite (NaOCl) is widely utilized in endodontics for its effective antimicrobial properties and ability to dissolve necrotic tissues. However, its use is not devoid of risks, including potential severe tissue damage and chemical burns. This systematic review evaluates the documented risks of using NaOCl in endodontic treatments. It compares its safety and efficacy with other endodontic irrigants, such as EDTA and chlorhexidine. This review followed the "Preferred Reporting Items for Systematic Reviews and Meta-Analyses" guidelines and registered with PROSPERO. Comprehensive searches were performed in PubMed, Scopus, Web of Science, and CENTRAL databases, with additional hand searches in grey literature. Studies were selected reporting adverse effects related to NaOCl used in endodontic procedures, ranging from randomized controlled trials to case reports. Data extraction and risk of bias assessment were systematically performed using standardized tools. Seventeen studies were included, detailing instances from chemical burns to severe allergic reactions and tissue necrosis associated with NaOCl use. The risk of bias was predominantly low across the studies. Sodium hypochlorite demonstrated a higher efficacy in microbial eradication and tissue dissolution than other irrigants, though it also showed a higher incidence of severe complications when mishandled. Sodium hypochlorite remains a cornerstone in endodontic disinfection due to its potent antimicrobial and tissue dissolution properties. However, its application must be meticulously managed to prevent complications. Future research should focus on optimizing concentrations and application techniques to enhance safety and effectiveness, potentially integrating safer alternatives or complementary solutions like EDTA to mitigate risks while preserving irrigant benefits.

Vesiculobullous Eruption due to Extravasation of Intravenous Fluid: A Brief Review

Abstract Introduction: Intravenous (IV) cannulation for peripheral or central venous access is a commonly done procedure in hospitals. Extravasation of IV fluids is one of the iatrogenic complications encountered. Extravasation injury may range from mild benign injury most of the time to edema, bullous lesion, severe tissue necrosis, and compartment syndrome. Methods: “Extravasation” and “Intravenous fluid” are searched in PubMed, PubMed Central, and Medline with articles published in the past 5 years. Few off topics such as “blood extravasation,” “extravasation injury to brain,” animal studies, and “extravasation of anticancer drugs” are excluded from the study. Twenty-one articles including review, meta-analysis, and case reports are selected for review. Results: While much of the focus is on peripheral venous access, for central venous catheters, extravasation is less frequent but potentially more dangerous because the anatomical structures escape attention. Along with mechanical factors such as volume of fluid administered and wrong placement of cannula, underlying diseases such as cancer, diabetes, and inherent property of IV fluid or infusion may be responsible for extravasation injury. Pain, paresthesia, pallor, and pulse to be examined and monitored while suspecting an extravasation injury. Ultrasonography and color Doppler might help to identify early lesions, but a high degree of clinical suspicion is necessary for unconscious patients in the intensive care unit. A detailed discussion on this topic is followed in this article. Conclusion: Patients should have their infusion site routinely inspected. The prevention of potentially fatal consequences depends on careful adherence to recommended procedures and prompt recognition of signs and symptoms.

Splenetic Torsion on Ectopic Spleen: A Case Report

Splenic volvulus, a rare torsion of the spleen around its vascular axis, can lead to severe tissue necrosis without prompt treatment. Underlying anatomical factors such as elongation of the splenic pedicle and ligament laxity increase the risk of torsion. Diagnosis is often made during emergency surgical intervention due to the rarity of the condition and the lack of specificity of symptoms. A 30-year-old woman presented with acute abdominal symptoms accompanied by fever, leading to emergency laparotomy revealing splenic volvulus. Imaging, particularly computed tomography, plays a crucial role in diagnosis. Treatment typically involves splenectomy in cases of necrosis. Wandering spleen, usually caused by ligament hyperlaxity, is a rare but important entity to recognize, often requiring surgical intervention to prevent serious complications.

Advances of Long Non-Coding RNAs as Potential Biomarkers for Tuberculosis: New Hope for Diagnosis?

Tuberculosis (TB), one of the top ten causes of death globally induced by the infection of Mycobacterium tuberculosis (Mtb), remains a grave public health issue worldwide. With almost one-third of the world's population getting infected by Mtb, between 5% and 10% of these infected individuals are predicted to develop active TB disease, which would not only result in severe tissue damage and necrosis, but also pose serious threats to human life. However, the exact molecular mechanisms underlying the pathogenesis and immunology of TB remain unclear, which significantly restricts the effective control of TB epidemics. Despite significant advances in current detection technologies and treatments for TB, there are still no appropriate solutions that are suitable for simultaneous, early, rapid, and accurate screening of TB. Various cellular events can perturb the development and progression of TB, which are always associated with several specific molecular signaling events controlled by dysregulated gene expression patterns. Long non-coding RNAs (lncRNAs), a kind of non-coding RNA (ncRNA) with a transcript of more than 200 nucleotides in length in eukaryotic cells, have been found to regulate the expression of protein-coding genes that are involved in some critical signaling events, such as inflammatory, pathological, and immunological responses. Increasing evidence has claimed that lncRNAs might directly influence the susceptibility to TB, as well as the development and progression of TB. Therefore, lncRNAs have been widely expected to serve as promising molecular biomarkers and therapeutic targets for TB. In this review, we summarized the functions of lncRNAs and their regulatory roles in the development and progression of TB. More importantly, we widely discussed the potential of lncRNAs to act as TB biomarkers, which would offer new possibilities in novel diagnostic strategy exploration and benefit the control of the TB epidemic.

Abstract 298: Roles Of Tet2 In Vascular Regeneration For Peripheral Artery Disease

Peripheral artery disease (PAD) is the manifestation of atherosclerosis in the extremity, causing blockage of blood vessels that reduces blood flow to the lower limb (ischemia). End-stage PAD patients require limb amputation, with increased risk of morbidity and mortality from heart attacks. Although PAD affects >200 million individuals worldwide, surgery remains the only endpoint option that itself is associated with significant morbidity and mortality. Novel treatment for PAD is therefore urgently needed. Despite vascular smooth muscle cells (VSMC) being essential for normal vascular formation and repair, their role in PAD pathogenesis has been largely ignored. Our lab has long been specialised in studying VSMC biology, and was the first to identify a DNA demethylase, Tet2, as a master regulator of VSMC functions ( Liu et al, Circulation. 2013 ). We found without Tet2, VSMCs form pathological and weakened arteries that lead to restenosis, the precursor of atherosclerosis. Given the vital role VSMC in vessel formation, we hypothesise that restoring Tet2 expression to ischaemic tissues is critical for promoting angiogenesis, blood flow recovery and to prevent tissue caused by PAD. Using a VSMC-specific Tet2 knockout (VSM- Tet2 KO ) mouse line generated by the Liu lab at VCCRI, we studied the vasculo-protective role of TET2 in PAD by mimicking PAD through femoral artery ligation. Our preliminary data demonstrates: (1) severe tissue necrosis and amputation in the knockout mice following surgery that is not observed in controls (2) Loss of Tet2 reduces blood flow to the foot as suggested by laser doppler measurements. (3) Significantly retarded neovascularisation in ischemic muscle regions displayed by micro-computer tomography scans . These are the first data to demonstrate Tet2’s role in PAD.

Impact of Colchicine on Histology of Testis in Rats

Colchicine is a drug widely used for the management of many disorders, such as acute gout and Behçet’s disease. It is also prescribed for the treatment of pericarditis, atrial fibrillation coronary artery diseases, and secondary amyloidosis. In case this drug is used at the early stages of coronavirus infection, its anti-inflammatory properties may reduce the severe inflammatory reactions related to a cytokine storm by affecting the inflammasome. The purpose of the present study was to determine the toxicity of Colchicine on testis in rats from different age groups for 10 days. A total of 27 male Wistar rats were divided into three groups. The rats in group I (control group) were administered distilled water by oral gavage. Group II consisted of young rats (5-6 months old) who orally received Colchicine 3 mg/kg body weight. Group III entailed rats of 14-16 months who were orally administered colchicine 3 mg/kg body weight. The testis of the treated groups was dissected and examined for histological changes and morphometrical analysis. The obtained results indicated that high doses of Colchicine (3 mg/kg body weight) could induce tissue damage to the testis, including degeneration and necrosis of both Sertoli and Leydig cells with irregular divisions of germinal epithelium, even when it was used for short periods (10 days). In the elderly treated rats, there were severe tissue damages, including degeneration and necrosis of germinal epithelium with irregular divisions of germ cells, necrosis of Sertoli and Leydig cells with sloughing of germinal epithelium toward the lumen of the tubule. Therefore, there is a need to conduct more studies to investigate the side effect of Colchicine as it is excessively used in the management of coronavirus.

Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by Naja atra Venom on Myoblast C2C12 Cells

The Taiwanese cobra, Naja atra, is a clinically significant species of snake observed in the wild in Taiwan. Victims bitten by N. atra usually experience severe pain and local tissue necrosis. Although antivenom is available for treatment of cobra envenomation, its neutralization potency against cobra-induced necrosis is weak, with more than 60% of cobra envenoming patients developing tissue necrosis after antivenom administration. The present study found that cytotoxin (CTX) is a key component of N. atra venom responsible for cytotoxicity against myoblast cells. Anti-CTX IgY was generated in hens, and the spleens of these hens were used to construct libraries for the development of single chain variable fragments (scFv). Two anti-CTX scFv, S1 and 2S7, were selected using phage display technology and biopanning. Both polyclonal IgY and monoclonal scFv S1 reacted specifically with CTX in cobra venom. In a cell model assay, the CTX-induced cytolytic effect was inhibited only by monoclonal scFv S1, not by polyclonal IgY. Moreover, the neutralization potency of scFv S1 was about 3.8 mg/mg, approximately three times higher than that of conventional freeze-dried neurotoxic antivenom (FNAV). Collectively, these results suggest that scFv S1 can effectively neutralize CTX-induced cytotoxicity and, when combined with currently available antivenom, can improve the potency of the latter, thereby preventing tissue damage induced by cobra envenoming.

Snakebites in Jordan: A clinical and epidemiological study

The epidemiology features and clinical symptoms associated with 121 cases of snakebite requiring admission to 24 hospitals in Jordan, during 2018–2020, are discussed. Ninety-six of them (79%) brought with them to the hospital the snake that bit them or a photograph of it. Echis coloratus was responsible for 68 of the bites and 6 fatalities. Sex ratio was 3.2 males: 1 female, with an overall average age of 27 ± 14.36 years. The highest incidence of bites was reported in September. Bites were most common on hands and legs. The period of hospitalization ranged from 1 to 36 days. Irbid and Karak governorates had the highest number of snakebites, most cases being reported from agricultural areas and among farmers.Clinical symptoms associated with five species of venomous snakes are described along with illustrative case histories. Symptoms associated with Echis coloratus bites included local swelling and necrosis, coagulopathy and bleeding, microangiopathic hemolytic anemia, thrombocytopenia, and acute kidney injury (AKI) and chronic renal failure. Daboia palaestinae victims exhibited ecchymoses, local swelling and necrosis, with one case of angioedema. Other symptoms included thrombocytopenia, coagulopathy, microangiopathic hemolysis and local and systemic bleeding, as well as AKI. A single case of envenoming by Macrovipera lebetinus developed the following symptoms; swelling, severe pain, extensive ecchymoses, neutrophil leukocytosis, normochromic normocytic anemia and aggregated platelets with thrombocytopenia. Symptoms associated with two cases of Pseudocerastes fieldi included swelling that spread from bitten hands, and mild abnormalities of platelet count and bleeding time. Four cases of envenoming by Atractaspis engaddensis exhibited severe pain, local swelling, erythema, numbness and tissue necrosis. One of them developed acute systemic symptoms.The only antivenom currently available in Jordan, is VINS “Snake venom antitoxin (Biosnake)”, manufactured in India using venoms of three African snakes that do not occur in Jordan. It proved clinically ineffective against envenoming by Jordanian Viperidae, failing to correct coagulopathy and life-threatening hemorrhage, and to prevent AKI.

Scale Drop Disease Virus (SDDV) and Lates calcarifer Herpes Virus (LCHV) Coinfection Downregulate Immune-Relevant Pathways and Cause Splenic and Kidney Necrosis in Barramundi Under Commercial Farming Conditions.

Marine farming of barramundi (Lates calcarifer) in Southeast Asia is currently severely affected by viral diseases. To better understand the biological implications and gene expression response of barramundi in commercial farming conditions during a disease outbreak, the presence of pathogens, comparative RNAseq, and histopathology targeting multiple organs of clinically “sick” and “healthy” juveniles were investigated. Coinfection of scale drop disease virus (SDDV) and L. calcarifer herpes virus (LCHV) were detected in all sampled fish, with higher SDDV viral loads in sick than in healthy fish. Histopathology showed that livers in sick fish often had moderate to severe abnormal fat accumulation (hepatic lipidosis), whereas the predominant pathology in the kidneys shows moderate to severe inflammation and glomerular necrosis. The spleen was the most severely affected organ, with sick fish presenting severe multifocal and coalescing necrosis. Principal component analysis (PC1 and PC2) explained 70.3% of the observed variance and strongly associated the above histopathological findings with SDDV loads and with the sick phenotypes, supporting a primary diagnosis of the fish being impacted by scale drop disease (SDD). Extracted RNA from kidney and spleen of the sick fish were also severely degraded likely due to severe inflammation and tissue necrosis, indicating failure of these organs in advanced stages of SDD. RNAseq of sick vs. healthy barramundi identified 2,810 and 556 differentially expressed genes (DEGs) in the liver and muscle, respectively. Eleven significantly enriched pathways (e.g., phagosome, cytokine-cytokine-receptor interaction, ECM-receptor interaction, neuroactive ligand-receptor interaction, calcium signaling, MAPK, CAMs, etc.) and gene families (e.g., tool-like receptor, TNF, lectin, complement, interleukin, chemokine, MHC, B and T cells, CD molecules, etc.) relevant to homeostasis and innate and adaptive immunity were mostly downregulated in sick fish. These DEGs and pathways, also previously identified in L. calcarifer as general immune responses to other pathogens and environmental stressors, suggest a failure of the clinically sick fish to cope and overcome the systemic inflammatory responses and tissue degeneration caused by SDD.

Partial Decellularization for Segmental Tracheal Scaffold Tissue Engineering: A Preliminary Study in Rabbits.

In this study, we developed a new procedure for the rapid partial decellularization of the harvested trachea. Partial decellularization was performed using a combination of detergent and sonication to completely remove the epithelial layers outside of the cartilage ring. The post-decellularized tracheal segments were assessed with vital staining, which showed that the core cartilage cells remarkably remained intact while the cells outside of the cartilage were no longer viable. The ability of the decellularized tracheal segments to evade immune rejection was evaluated through heterotopic implantation of the segments into the chest muscle of rabbits without any immunosuppressive therapy, which demonstrated no evidence of severe rejection or tissue necrosis under H&E staining, as well as the mechanical stability under stress-pressure testing. Finally, orthotopic transplantation of partially decellularized trachea with no immunosuppression treatment resulted in 2 months of survival in two rabbits and one long-term survival (2 years) in one rabbit. Through evaluations of posttransplantation histology and endoscopy, we confirmed that our partial decellularization method could be a potential method of producing low-immunogenic cartilage scaffolds with viable, functional core cartilage cells that can achieve long-term survival after in vivo transplantation.

Traumatic Panniculitis-the Spectrum

Panniculitis, when induced by physical trauma or by chemical agents at injection sites, presents as indurated subcutaneous nodules or plaques. The clinical picture may vary, but the context makes usually the diagnosis easy. Three case histories from our department illustrate the spectrum of traumatic panniculitis: subcutaneous nodules at injection sites, a large subcutaneous mass disproportionate to mild trauma, and, at the other end of the spectrum, severe adipose tissue necrosis with liquefied fat discharging through surface wounds. Traumatic panniculitis is self-limiting and only requires symptomatic treatment.

Echis carinatus snake venom metalloprotease-induced toxicities in mice: Therapeutic intervention by a repurposed drug, Tetraethyl thiuram disulfide (Disulfiram).

Echis carinatus (EC) is known as saw-scaled viper and it is endemic to the Indian subcontinent. Envenoming by EC represents a major cause of snakebite mortality and morbidity in the Indian subcontinent. Zinc (Zn++) dependent snake venom metalloproteases (SVMPs) present in Echis carinatus venom (ECV) is well known to cause systemic hemorrhage and coagulopathy in experimental animals. An earlier report has shown that ECV activates neutrophils and releases neutrophil extracellular traps (NETs) that blocks blood vessels leading to severe tissue necrosis. However, the direct involvement of SVMPs in the release of NETs is not clear. Here, we investigated the direct involvement of EC SVMPs in observed pathological symptoms in a preclinical setup using specific Zn++ metal chelator, Tetraethyl thiuram disulfide (TTD)/disulfiram. TTD potently antagonizes the activity of SVMPs-mediated ECM protein degradation in vitro and skin hemorrhage in mice. In addition, TTD protected mice from ECV-induced footpad tissue necrosis by reduced expression of citrullinated H3 (citH3) and myeloperoxidase (MPO) in footpad tissue. TTD also neutralized ECV-induced systemic hemorrhage and conferred protection against lethality in mice. Moreover, TTD inhibited ECV-induced NETosis in human neutrophils and decreased the expression of peptidyl arginine deiminase (PAD) 4, citH3, MPO, and p-ERK. Further, we demonstrated that ECV-induced NETosis and tissue necrosis are mediated via PAR-1-ERK axis. Overall, our results provide an insight into SVMPs-induced toxicities and the promising protective efficacy of TTD can be extrapolated to treat severe tissue necrosis complementing anti-snake venom (ASV).

Gas Gangrene in a Closed Pelvic Injury

Gas gangrene in a closed pelvic injury is very rare phenomenon and we could not find many reported cases in the literature. Radiographs are not enough for diagnosis and computed tomography (CT) is not widely available yet in primary health center. Basic management includes surgical debridement with incision and drainage of necrotic tissue of the involved area, antibiotic therapy, and surgical intensive care. Delay of the surgical debridement for >12 h is associated with higher overall morbidity. The aerobic and anaerobic bacteria with the indigenous commensal of the pelvis results in the production of exotoxins that leads to severe rapid tissue necrosis and the synthesis of insoluble gases characteristic of subcutaneous emphysema or Fournier's gangrene. Although gas gangrene is often diagnosed clinically, emergency CT can support in early diagnosis with accurate assessment of the disease extent. CT not only helps evaluate the pelvic bony structures but also helps assess the spread of disease in the abdomen and pelvis. The mainstay of the management includes surgical debridement with incision and drainage of necrotic tissue of involved area, antibiotic therapy, and surgical intensive care. Early diagnosis remains the mainstay of the treatment of Gas gangrene. It is important to diagnose these life-threatening conditions and treat them as early as possible. Due to the rarity of such events and the unavailability of gold standard diagnostic and treatment modalities, it is very difficult to prevent the mortality for such patients. There is a need for a better treatment plan to diagnose and manage such patients urgently and not to miss the initial phase of disease progression which is critical for saving the life of the patient.

Hmox1 (Heme Oxygenase-1) Protects Against Ischemia-Mediated Injury via Stabilization of HIF-1α (Hypoxia-Inducible Factor-1α).

Hmox1 (heme oxygenase-1) is a stress-induced enzyme that catalyzes the degradation of heme to carbon monoxide, iron, and biliverdin. Induction of Hmox1 and its products protect against cardiovascular disease, including ischemic injury. Hmox1 is also a downstream target of the transcription factor HIF-1α (hypoxia-inducible factor-1α), a key regulator of the body's response to hypoxia. However, the mechanisms by which Hmox1 confers protection against ischemia-mediated injury remain to be fully understood. Approach and Results: Hmox1 deficient (Hmox1-/-) mice had impaired blood flow recovery with severe tissue necrosis and autoamputation following unilateral hindlimb ischemia. Autoamputation preceded the return of blood flow, and bone marrow transfer from littermate wild-type mice failed to prevent tissue injury and autoamputation. In wild-type mice, ischemia-induced expression of Hmox1 in skeletal muscle occurred before stabilization of HIF-1α. Moreover, HIF-1α stabilization and glucose utilization were impaired in Hmox1-/- mice compared with wild-type mice. Experiments exposing dermal fibroblasts to hypoxia (1% O2) recapitulated these key findings. Metabolomics analyses indicated a failure of Hmox1-/- mice to adapt cellular energy reprogramming in response to ischemia. Prolyl-4-hydroxylase inhibition stabilized HIF-1α in Hmox1-/- fibroblasts and ischemic skeletal muscle, decreased tissue necrosis and autoamputation, and restored cellular metabolism to that of wild-type mice. Mechanistic studies showed that carbon monoxide stabilized HIF-1α in Hmox1-/- fibroblasts in response to hypoxia. Our findings suggest that Hmox1 acts both downstream and upstream of HIF-1α, and that stabilization of HIF-1α contributes to Hmox1's protection against ischemic injury independent of neovascularization.

The consequences of delayed treatment in doxorubicin extravasation

Doxorubicin (Adriamycin) is an anthracycline chemotherapy agent used for the treatment of several cancers including haematological malignancies. Extravasation is a well-recognized and possibly serious complication of administered intravenous therapies, especially in chemotherapeutic agents. Drug extravasation into the surrounding tissue has been estimated to occur in 0.1–6% of patients that receive chemotherapy. Signs of the injury can be subtle, and in case of delayed detection (> 6 h) and treatment, doxorubicin extravasation can result in substantial tissue damage with significant loss of function depending on the location of injury. Other consequences include pain, physical defect, psychological distress, cost of hospitalization and treatment and more extensive damage to skin and underlying tissue. In this article, we will discuss a case of delayed treatment of doxorubicin extravasation from a Venflon venous line at the dorsum of the hand with severe tissue necrosis as a result. Successful surgical reconstruction of the resulting substantial tissue damage using a SCIAP flap 25 days following the extravasation is described. Level of evidence: Level V, risk study.

Cordycepin protects against acute pancreatitis by modulating NF-κB and NLRP3 inflammasome activation via AMPK

Acute pancreatitis (AP) is a noninfectious inflammatory disease with high morbidity and mortality, which is characterized by severe inflammation and tissue necrosis. Cordycepin (CRD), derived from Cordyceps militaris, possesses anti-inflammatory effects and immunomodulation properties. Here, we investigated the protective effects of CRD on pancreatic injury and clarified potential mechanisms in AP model. There were established caerulein-induced AP and CRD pretreatment models in vivo and in vitro, as showed by serum enzymes, histopathological alterations and pro-inflammatory cytokines. Pretreatment with CRD notably downregulated the serum amylase and lipase levels and apparently reduced pancreatic histopathological alterations in AP mice. Meanwhile, the MPO staining confirmed that CRD pretreatment modulated the infiltration of neutrophils in AP mice. Furthermore, CRD markedly decreased the levels of pro-inflammatory factors (IL-6, IL-1β, and TNF-α) though inhibiting the activation of nuclear factor-κB (NF-κB) and NLR family pyrin domain-containing protein 3 (NLRP3) inflammasome in AP mice. In pancreatic acinar cancer cell 266-6, CRD pretreatment decreased cholecystokinin(CCK)-induced inflammatory response was consistent with those in vivo. Mechanistically, CRD was also revealed to activate activated protein kinase (AMPK) and attenuated inflammation both in vivo and in vitro. On the whole, this study indicated that CRD protects mice from pancreatic inflammatory process and damage by suppressed NF-κB and NLRP3 inflammasome activation via AMPK, which probably contributed to the potential therapy for AP.

Intratumoral injection of dendritic cells overexpressing interleukin‑12 inhibits melanoma growth.

The present study aimed to investigate the antitumor effects of an intratumoral injection of dendritic cells (DCs) overexpressing interleukin-12 (IL‑12) on melanoma. DCs, isolated from mouse spleen, were gene‑modified using an IL‑12 overexpression vector. Melanoma B6 cells were injected into C57BL/6 mice to generate tumors. Thereafter, DCs overexpressing IL‑12 were injected into the tumors, and tumor volume was subsequently measured. Pathological changes in tumor tissue were detected by hematoxylin and eosin staining. The expression of interleukin-4 (IL‑4) and IL‑12 in tumors was measured by enzyme‑linked immunosorbent assay, real‑time PCR and western blotting. DCs were successfully isolated and a lentivirus vector expressing IL‑12 was constructed. After intratumoral injection of phosphate‑buffered saline (control group), tumor cells exhibited malignant growth; whereas tumors injected with DCs (DC group) or DCs + empty vector (DC + vector group) exhibited a small amount of inflammatory cell infiltration and limited areas of tissue necrosis. In contrast, tumors injected with DCs overexpressing IL‑12 (DC + IL‑12 group) displayed severe tissue necrosis, loss of cell structure, and inflammatory cell infiltration. Compared with the control group, the tumor volumes were significantly lower in the DC, the DC + vector and the DC + IL‑12 groups, while the expression of IL‑12 and IL‑4 in the tumors was significantly higher. Importantly, the most marked changes in tumor volume and IL‑12 and IL‑4 expression were in the DC + IL‑12 group, which were significantly greater than those in tumors treated with unmodified DCs. Hence, intratumoral injection of DCs overexpressing IL‑12 exerted strong antitumor effects in melanoma, and biotherapy with DCs overexpressing IL‑12 is a potential treatment strategy for melanoma.

Capsular polysaccharide of Streptococcus agalactiae is an essential virulence factor for infection in Nile tilapia (Oreochromis niloticus Linn.).

Streptococcus agalactiae (Group B Streptococcus, GBS) is associated with diverse diseases in aquatic animals. The capsule polysaccharide (CPS) encoded by the cps gene cluster is the major virulence factor of S.agalactiae; however, limited information is available regarding the pathogenic role of the CPS of serotype Ia piscine GBS strains in fish. Here, a non-encapsulated mutant (Δcps) was constructed by insertional mutagenesis of the cps gene cluster. Mutant pathogenicity was evaluated in vitro based on the killing of whole blood from tilapia, in vivo infections, measuring mutant survival in tilapia spleen tissues and pathological analysis. Compared to wild-type (WT) GBS strain, the Δcps mutant had lower resistance to fresh tilapia whole blood in vitro (p<0.01), and more easily cleared in tilapia spleen tissue, and was highly attenuated in tilapia and zebrafish. Additionally, compared to the Δcps mutant, numerous GBS strains and severe tissue necrosis were observed in the tilapia spleen tissue infected with WT strains. These results indicated that the CPS is essential for GBS pathogenicity and may serve as a target for attenuation in vaccine development. Gaining a better understanding of the role, the GBS pathogenicity in fish will provide insight into related pathogenesis and host-pathogen interactions.

Plant DNases are potent therapeutic agents against Echis carinatus venom-induced tissue necrosis in mice.

Echis carinatus envenomation leads to severe tissue necrosis at the bitten site by releasing DNA from immune cells that blocks the blood flow. An earlier report has shown that exogenous DNase 1 offers protection against such severe local tissue necrosis. Tricosanthus tricuspidata is a medicinal plant and the paste prepared from its leaves has been used extensively for the treatment of snakebite-induced tissue necrosis. Most studies including reports from our laboratory focused on plant secondary metabolite as therapeutic molecules against snakebite envenomation. However, the involvement of hydrolytic enzymes including DNase in treating snake venom-induced tissue necrosis has not been addressed. Several folk medicinal plants used against snakebite treatment showed the presence of DNase activity and found to be rich in T.tricuspidata. Further, purified T.tricuspidata DNase showed a single sharp peak in reversed-phase high-performanceliquid chromatography(RP-HPLC) with an apparent molecular mass of 17 kDa. T.tricuspidata DNase exhibited potent DNA degrading activity performed using agarose gel electrophoresis, spectrophotometric assay, and DNA zymography. In addition, purified DNase from T.tricuspidata was able to neutralize E.carinatus venom-induced mouse tail tissue necrosis and normalized elevated serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels 30 minutes post venom injection. T.tricuspidata DNase was also able to reverse E.carinatus venom-induced histopathological changes and collagen depletion in mice tail tissue. All these observed pharmacological actions of T.tricuspidata DNase were inhibited by sodium fluoride (NaF). This study provides scientific validation of the traditional use of T.tricuspidata leaf paste in the healing of snakebite-induced tissue necrosis and might be exploited to treat snake venom-induced local toxicity.

Case Challenge: 70-Year Old Female Who Presented With a 3-Month Non-Healing Leg Wound

Pyoderma gangrenosum (PG) is an uncommon chronic and progressive skin disorder that can lead to severe tissue necrosis, pathergy, horrendous pain, and disfigurement if not properly and promptly diagnosed and treated. The presentation was based on previous case report on a patient with PG who initially presented with a history of a painful lower leg wound, presumed to be caused by venous disease.1 Following 3 months of appropriate systemic treatments, topical wound care, and limited progress, a dehydrated human amnion/chorion membrane allograft was applied to the wound. The purpose of this case study is to describe the care of a patient with painful PG whose wound was managed with Dehydrated Human Amnion/Chorion Membrane (dHACM) as part of her treatment plan.