Spatial Mass Cytometry-Based Single-Cell Imaging Reveals a Disrupted Epithelial-Immune Axis in Prurigo Nodularis.

Prurigo nodularis (PN) is a chronic, inflammatory skin condition that disproportionately affects African Americans and features intensely pruritic, hyperkeratotic nodules on the extremities and trunk. PN is understudied compared with other inflammatory skin diseases, with the spatial organization of the cutaneous infiltrate in PN yet to be characterized. In this work, we employ spatial imaging mass cytometry to visualize PN lesional skin inflammation and architecture with single-cell resolution through an unbiased machine learning approach. PN lesional skin has increased expression of caspase 3, NF-kB, and phosphorylated signal transducer and activator of transcription 3 compared with healthy skin. Keratinocytes in lesional skin are subdivided into CD14+CD33+, CD11c+, CD63+, and caspase 3-positive innate subpopulations. CD14+ macrophage populations expressing phosphorylated extracellular signal-regulated kinase 1/2 correlate positively with patient-reported itch (P = .006). Hierarchical clustering reveals a cluster of patients with PN with greater atopy, increased NF-kB+ signal transducer and activator of transcription 3-positive phosphorylated extracellular signal-regulated kinase 1/2-positive monocyte-derived myeloid dendritic cells, and increased vimentin expression (P < .05). Neighborhood analysis finds interactions between CD14+ macrophages, CD3+ T cells, monocyte-derived myeloid dendritic cells, and keratinocytes expressing innate immune markers. These findings highlight phosphorylated extracellular signal-regulated kinase-positive CD14+ macrophages as contributors to itch and suggest an epithelial-immune axis in PN pathogenesis.

Topical GZ21T Inhibits the Growth of Actinic Keratoses in a UVB-Induced Model of Skin Carcinogenesis.

Actinic keratoses (AKs) are premalignant intraepidermal neoplasms that occur as a result of cumulative sun damage. AKs commonly relapse, and up to 16% undergo malignant transformation into cutaneous squamous cell carcinoma. There is a need for novel therapies that reduce the quantity and surface area of AKs as well as prevent malignant transformation to cutaneous squamous cell carcinomas. We recently showed that GZ17-6.02, an anticancer agent composed of curcumin, haramine, and isovanillin, inhibited the growth of H297.T cells. This study evaluated the efficacy of a topical formulation of GZ17-6.02, known as GZ21T, in a murine model of AK generated by exposing SKH1 mice to UVR. Treatment of mice with topical GZ21T inhibited the growth of AKs by decreasing both lesion count (P = 0.012) and surface area occupied by tumor (P = 0.002). GZ21T also suppressed the progression of AKs to cutaneous squamous cell carcinoma by decreasing the count (P = 0.047) and surface area (P = 0.049) of lesions more likely to represent cutaneous squamous cell carcinoma. RNA sequencing and proteomic analyses revealed that GZ21T suppressed several pathways, including MAPK (P = 0.025), phosphoinositide 3-kinase-protein kinase B (P = 0.04), HIF-1α (P = 0.016), Wnt (P = 0.025), insulin (P = 0.018), and ERBB (P = 0.016) signaling. GZ21T also upregulated the autophagy-promoting protein AMPK while suppressing proteins such as PD-L1, glutaminase, pAkt1 S473, and eEF2K.

An oral caspase inhibitor as monotherapy or with antibiotics eradicates MRSA skin infections in mice.

Staphylococcus aureus is the leading cause of skin and soft tissue infections. With the emergence of antibiotic-resistant bacteria, there is an unmet clinical need to develop immune-based therapies to treat skin infections. Previously, we have shown pan-caspase inhibition as a potential host-directed immunotherapy against community-acquired methicillin-resistant S aureus (CA-MRSA) and other bacterial skin infections. Here, we evaluated the role of irreversible pan-caspase inhibitor emricasan as a monotherapy and an adjunctive with a standard-of-care antibiotic, doxycycline, as potential host-directed immunotherapies against S. aureus skin infections in vivo. We used the established CA-MRSA strain USA300 on the dorsum of WT C57BL/6J mice and monitored lesion size and bacterial burden noninvasively, and longitudinally over 14 days with in vivo bioluminescence imaging (BLI). Mice in four groups placebo (0.5% carboxymethyl cellulose [CMC] solution), placebo plus doxycycline (100 mg/kg), emricasan (40 mg/kg) plus doxycycline, and emricasan only were treated orally twice daily by oral gavage for 7 days, starting at 4 h after injection of S aureus. When compared with placebo, all three groups, placebo plus doxycycline, emricasan plus doxycycline, and emricasan treated group, exhibited biological effect, with reduction of both the lesion size (*p = .0277, ****p < .0001, ****p < .0001, respectively) and bacterial burden (***p = .003, ****p < .0001, ****p < .0001, respectively). Importantly, the efficacy of emricasan against S. aureus was not due to direct antibacterial activity. Collectively, pan-caspase inhibitor emricasan and emricasan plus doxycycline reduced both the lesion size and bacterial burden in vivo, and emricasan is a potential host-directed immunotherapy against MRSA skin infections in a preclinical mouse model.

Reduced serum pyridoxine and 25-hydroxyvitamin D levels in adults with chronic pruritic dermatoses.

Little is known about the role nutritional factors play in the pathogenesis of chronic pruritic dermatoses (CPD). In this study, we analyzed nutritional deficiencies in CPD patients compared to matched controls. We conducted a population-based study from the National Health and Nutrition Examination Survey (NHANES) from 2005 to 2006. The main outcomes of the study were laboratory data on serum vitamin levels in participants who answered affirmatively to the questionnaires on CPD as well as matched healthy controls. We identified 877 cases of CPD among 9817 adults in the US aged 20 to 59 years. These findings revealed a slightly higher percentage of females with CPD. Low vitamin B6 (OR 0.697; 95% CI: 0.696-0.699, p = 0.025) and vitamin D (OR 0.794; 95% CI: 0.789-0.799, p = 0.037) levels were associated with a higher rate of CPD compared to healthy controls. Our study suggests that low levels of Vitamin B6 and Vitamin D inversely correlates with the presence of CPD. These vitamin deficiencies suggest further studies on the effect of vitamin supplementation may help in patients with CPD.

Risk of Hematologic Cancer in Patients With Undifferentiated Pruritus.

Importance: Although pruritus is common in patients with hematologic cancers, it is unknown whether patients with undifferentiated pruritus have higher risk of developing hematologic cancer. Furthermore, it is unclear whether serum lactate dehydrogenase (LDH) level, commonly ordered for cancer workup, has diagnostic utility in patients with pruritus. Objective: To assess the risk of hematologic cancer and the diagnostic utility of LDH level in patients with undifferentiated pruritus. Design, Setting, and Participants: This retrospective population-level cohort analysis was conducted using the TriNetX Research Network, a global health records database encompassing more than 69 million patients, from 2002 to 2020. The study included 327 502 eligible patients diagnosed with unspecified pruritus, excluding those with existing chronic pruritic dermatoses or systemic diseases known to cause pruritus, along with 327 502 matched controls. Exposures: Development of hematologic cancer within 1 year, 5 years, and 10 years following unspecified pruritus diagnosis. Main Outcomes and Measures: Primary study outcomes were 1-year, 5-year, and 10-year relative risks (RRs) for development of 9 hematologic cancers in patients with pruritus compared with control patients. Secondary outcomes were 1-year, 5-year, and 10-year RRs for any hematologic cancer at different LDH cutoffs (250 U/L and 500 U/L). Results: After matching, the pruritus and control cohorts each had 327 502 patients (68.1% female patients; 0.4% American Indian or Alaska Native patients; 3.5% Asian patients; 22.2% Black patients; 0.1% Native Hawaiian or Pacific Islander patients; 59.3% White patients; mean [SD] age, 42.2 [22] years). Patients with pruritus had increased 1-year risk of Hodgkin lymphoma (RR, 4.42; 95% CI, 2.83-6.88), myeloid leukemia (RR, 2.56; 95% CI, 1.79-3.67), multiple myeloma (RR, 2.38; 95% CI, 1.66-3.41), non-Hodgkin lymphoma (RR, 2.35; 95% CI, 1.96-2.82), monoclonal gammopathy (RR, 1.90; 95% CI, 1.55-2.32), myelodysplastic syndrome (RR, 1.74; 95% CI, 1.14-2.64), and lymphocytic leukemia (RR, 1.47; 95% CI, 1.07-2.02). After 12 months, the cancer risk was comparable with that of controls. Patients with pruritus had increased LDH levels, which were not associated with increased hematologic cancer risk. Conclusions and Relevance: In this cohort study, the RR of hematologic cancer in patients with undifferentiated pruritus was highest in the first 12 months, and LDH level had limited diagnostic utility in these patients. Clinicians should consider a thorough review of symptoms and assessment of cancer risk factors when deciding on workup for patients presenting with undifferentiated pruritus.

Extracellular matrix and dermal nerve growth factor dysregulation in prurigo nodularis compared to atopic dermatitis.

Prurigo nodularis (PN) is a chronic, pruritic, inflammatory skin disease characterized by hyperkeratotic nodules on the trunk and extremities. While there is growing research on the immunological basis of PN, the neuropathic and structural components of PN lesions are unknown. This study examines the inflammatory, neuropathic, and structural pathways in PN compared to atopic dermatitis (AD) using RNA-sequencing of the lesional and non-lesional skin tissue of PN and AD patients, as well as immunohistochemistry analysis of nerve growth factor (NGF), a neurotrophic factor that regulates nerve development. Transcriptomic analysis of skin biopsies revealed that compared to lesional AD skin, lesional PN skin had significantly increased expression of NGF, matrix metalloproteinases, OSM, MCEMP1, IL1α, IL1ß, CXCL2, CXCL5, CXCL8, and insulin-like growth factors in PN compared to AD, and decreased expression of CCL13, CCL26, EPHB1, and collagens (COL4/6). Gene set enrichment analysis demonstrated higher enrichment of keratinization, cornified envelope, myelin sheath, TGF-beta signaling, extracellular matrix disassembly, metalloendopeptidase activity, and neurotrophin-TRK receptor signaling pathways in PN. On immunohistochemistry, PN lesions demonstrated higher dermal NGF expression compared to AD. We present novel findings demonstrating increased neurotrophic and extracellular matrix remodeling signatures in PN compared to AD, possibly explaining the morphological differences in their lesions. These signatures may therefore be important components of the PN pathogenesis and may serve as therapeutic targets.

Lead Toxicoses in Free-Range Chickens in Artisanal Gold-Mining Communities, Zamfara, Nigeria.

BACKGROUND: In early 2010, outbreaks of lead poisoning due to artisanal gold mining in villages in the northwest Nigerian state of Zamfara have resulted in the death of hundreds of children < 5 years old. There have also been unconfirmed reports of high mortality of geese within these villages. OBJECTIVES: To report a case of lead poisoning in three domestic free-range chickens found in one of the affected communities where illegal small-scale gold mining activities take place. METHODS: Three free-range domestic chickens were presented during a field investigation in one of the villages. The birds were observed to be emaciated, weak, showing nervous manifestations and moribund. RESULTS: Tissue extracts of liver, spleen and intestines were negative for Newcastle viral antigens, while cultures of liver and spleen biopsy were positive for Escherichia coli. Histopathological lesions were observed in the kidney, proventriculus and brain. Concentrations of lead in the tissues ranged between 7.5 mg/kg and 120.5 mg/kg wet weight, and the potential daily intake of lead in the tissues were estimated at 34.06-200.15 µg/day/kg body weight with an average of 118.37 µg/day/kg body weight. CONCLUSIONS: The results of the present study suggest probable risk to human health due to the consumption of chicken contaminated by lead in the affected villages. Poisoning in animal populations may serve as a sentinel to assess the extent of environmental contamination and human health problems related to lead. ETHICS APPROVAL: Protocols were approved and performed in accordance with relevant local guidelines and regulations as set by the Animal Care and Use Committee of the National Veterinary Research Institute, Vom, Nigeria. COMPETING INTERESTS: The authors declare no competing financial interests.

Anticoccidial effects of Khaya senegalensis aqueous stem bark extract on broiler chickens experimentally infected with Eimeria species.

Graded concentrations (200, 400 and 800 mg/kg) of the aqueous stem bark extract of Khaya senegalensis was evaluated for its therapeutic efficacy against experimentally induced coccidiosis in broiler chicken. The phytochemical analysis shows the presence of tannins, saponins, cardiac glycosides and steroids. There was significant reduction in oocyst count across the groups in a graded dose manner with 800 mg/kg being the most efficacious dose. There was also weight gain across the treatment groups with immuno-modulatory and erythropoetic activities observed. Also, a significant (p < 0.05) graded dose-dependent reduction in the oocyst count in the treatment groups. A significant (p < 0.05) increase in mean weight gain was also recorded across the experimental groups except the negative control. The haematology also showed a dose-dependent increase in red blood cells, haemoglobin and packed cell volume of the treatment groups. The extract had no significant difference (p > 0.05) on the white blood cells, but a slight decrease in the white blood cells and heterophil counts was observed at 400 mg/kg. Furthermore, the aspartate amino transaminase level showed a significant difference (p < 0.05). Fluctuating levels of other serum biochemical parameters such as total protein, albumin and potassium were observed. No significant difference (p > 0.05) in the sodium concentration was observed. In addition, oxidative stress biomarkers such as catalase significantly increased (p < 0.05) in all the experimental groups in addition to the concomitant increase in reduced gluthathione (GSH) and superoxide dismutase (SOD) levels. Conclusively, the aqueous extract of K. senegalensis was effective in the management of coccidiosis thus supporting its folkloric use.

Phosphine poisoning in free-range local chickens: a case report.

Phosphine poisoning is rarely reported in poultry, and its diagnosis is a great challenge for veterinary toxicologists and pathologists. A case of phosphine toxicosis in local, free range chickens is reported. Fourteen dead chickens (age ≥6 months old) were presented to the veterinary clinic for necropsy. The history revealed that the chickens were normally fed with guinea corn grains, but were suspected to have been fed boiled rice laced with poison. The clinical signs observed were ruffled feathers, somnolence, anorexia and high mortality. The disease ran a 2-day course with mortality pattern of four chickens the first day, six overnight and 14 the following day. Necropsy findings showed generalized vascular congestion and haemorrhage in the lungs and visceral organs, with the crop and gizzard filled with guinea corn and rice grains and greenish-yellow faecal material in the intestinal lumen. The presence of widespread congestion and petechial haemorrhages on visceral organs with the microscopic pulmonary congestion, and diffuse intraparabronchial presence of air sac macrophages, strongly suggested an acute toxic cause of death. Chemical tests on the crop contents of the dead chickens were positive for phosphine gas. This report will contribute to a better understanding of the clinical signs and lesions presented in cases of acute phosphide rodenticide exposure in domestic chickens, with a brief review of the forensic literature.

Phytochemical analysis and in-vitro anti-African swine fever virus activity of extracts and fractions of Ancistrocladus uncinatus, Hutch and Dalziel (Ancistrocladaceae).

BACKGROUND: African swine fever (ASF), a highly contagious fatal acute haemorrhagic viral disease of pigs currently has no treatment or vaccination protocol and it threatens the pig industry worldwide. Recent outbreaks were managed by farmers with ethnoveterinary preparations with various claims of effectiveness. RESULTS: We identified 35 compounds using GC-MS protocol and ASF virus (NIG 99) was significantly reduced by some extracts and fractions of the plant. However, the plant was poorly extracted by water and cytotoxicity was found to be a major problem with the use of the plant since its extracts also reduced the primary cells used in the assay. CONCLUSION: It is confirmed that the plant has antiviral potentials against ASF virus and farmers' claims seem to have certain degree of veracity, but finding the best means of exploring the potential of the plant while reducing its cytotoxic effect in-vitro and in-vivo will be necessary.

Ethnobotanical survey of medicinal plants used in the treatment of animal diarrhoea in Plateau State, Nigeria.

BACKGROUND: The use of medicinal plants in the treatment of diseases has generated renewed interest in recent times, as herbal preparations are increasingly being used in both human and animal healthcare systems. Diarrhoea is one of the common clinical signs of gastrointestinal disorders caused by both infectious and non-infectious agents and an important livestock debilitating condition. Plateau State is rich in savannah and forest vegetations and home to a vast collection of plants upheld in folklore as having useful medicinal applications. There is however scarcity of documented information on the medicinal plants used in the treatment of animal diarrhoea in the state, thus the need for this survey. Ten (10) out of 17 Local Government Areas (LGAs), spread across the three senatorial zones were selected. Farmers were interviewed using well structured, open-ended questionnaire and guided dialogue techniques between October and December 2010. Medicinal plants reported to be effective in diarrhoea management were collected using the guided field-walk method for identification and authentication. RESULTS: A total of 248 questionnaires were completed, out of which 207 respondents (83.47%) acknowledged the use of herbs in diarrhoea management, while 41 (16.53%) do not use herbs or apply other traditional methods in the treatment of diarrhoea in their animals. Medicinal plants cited as beneficial in the treatment of animal diarrhoea numbered 132, from which 57(43.18%) were scientifically identified and classified into 25 plant families with the families Fabaceae (21%) and Combretaceae (14.04%) having the highest occurrence. The plant parts mostly used in antidiarrhoeal herbal preparations are the leaves (43.86%) followed by the stem bark (29.82%). The herbal preparations are usually administered orally. CONCLUSION: Rural communities in Plateau State are a rich source of information on medicinal plants as revealed in this survey. There is need to scientifically ascertain the authenticity of the claimed antidiarrhoeal properties of these plants and perhaps develop more readily available alternatives in the treatment of diarrhoea.