Outbreak of Diarrhea Caused by a Novel Cryptosporidium hominis Subtype During British Military Training in Kenya.

Background: We report clinical, epidemiological, and laboratory features of a large diarrhea outbreak caused by a novel Cryptosporidium hominis subtype during British military training in Kenya between February and April 2022. Methods: Data were collated from diarrhea cases, and fecal samples were analyzed on site using the multiplex polymerase chain reaction (PCR) BioFire FilmArray. Water was tested using Colilert kits (IDEXX, UK). DNA was extracted from feces for molecular characterization of Cryptosporidium A135, Lib13, ssu rRNA, and gp60 genes. Results: One hundred seventy-two of 1200 (14.3%) personnel at risk developed diarrhea over 69 days. One hundred six primary fecal samples were tested, and 63/106 (59.4%; 95% CI, 0.49%-0.69%) were positive for Cryptosporidium spp. Thirty-eight had Cryptosporidium spp. alone, and 25 had Cryptosporidium spp. with ≥1 other pathogen. A further 27/106 (25.5%; 95% CI, 0.18%-0.35%) had non-Cryptosporidium pathogens only, and 16/106 (15.1%; 95% CI, 0.09%-0.23%) were negative. C. hominis was detected in 58/63 (92.1%) Cryptosporidium spp.-positive primary samples, but the others were not genotypable. Twenty-seven C. hominis specimens were subtypable; 1 was gp60 subtype IeA11G3T3, and 26 were an unusual subtype, ImA13G1 (GenBank accession OP699729), supporting epidemiological evidence suggesting a point source outbreak from contaminated swimming water. Diarrhea persisted for a mean (SD) of 7.6 (4.6) days in Cryptosporidium spp. cases compared with 2.3 (0.9) days in non-Cryptosporidium cases (P = .001). Conclusions: Real-time multiplex PCR fecal testing was vital in managing this large cryptosporidiosis outbreak. The etiology of a rare C. hominis gp60 subtype emphasizes the need for more genotypic surveillance to identify widening host and geographic ranges of novel C. hominis subtypes.

Costimulation of γδTCR and TLR7/8 promotes Vδ2 T-cell antitumor activity by modulating mTOR pathway and APC function.

BACKGROUND: Gamma delta (γδ) T cells are attractive effector cells for cancer immunotherapy. Vδ2 T cells expanded by zoledronic acid (ZOL) are the most commonly used γδ T cells for adoptive cell therapy. However, adoptive transfer of the expanded Vδ2 T cells has limited clinical efficacy. METHODS: We developed a costimulation method for expansion of Vδ2 T cells in PBMCs by activating γδ T-cell receptor (γδTCR) and Toll-like receptor (TLR) 7/8 using isopentenyl pyrophosphate (IPP) and resiquimod, respectively, and tested the functional markers and antitumoral effects in vitro two-dimensional two-dimensional and three-dimensional spheroid models and in vivo models. Single-cell sequencing dataset analysis and reverse-phase protein array were employed for mechanistic studies. RESULTS: We find that Vδ2 T cells expanded by IPP plus resiquimod showed significantly increased cytotoxicity to tumor cells with lower programmed cell death protein 1 (PD-1) expression than Vδ2 T cells expanded by IPP or ZOL. Mechanistically, the costimulation enhanced the activation of the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (PKB/Akt)-the mammalian target of rapamycin (mTOR) pathway and the TLR7/8-MyD88 pathway. Resiquimod stimulated Vδ2 T-cell expansion in both antigen presenting cell dependent and independent manners. In addition, resiquimod decreased the number of adherent inhibitory antigen-presenting cells (APCs) and suppressed the inhibitory function of APCs by decreasing PD-L1 and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) expression in these cells during in vitro Vδ2 T-cell expansion. Finally, we showed that human Vδ2 T cells can be expanded from PBMCs and spleen of humanized NSG mice using IPP plus resiquimod or ZOL, demonstrating that humanized mice are a promising preclinical model for studying human γδ T-cell development and function. CONCLUSIONS: Vδ2 T cells expanded by IPP and resiquimod demonstrate improved anti-tumor function and have the potential to increase the efficacy of γδ T cell-based therapies.

Neural Crest-Like Stem Cell Transcriptome Analysis Identifies LPAR1 in Melanoma Progression and Therapy Resistance.

Metastatic melanoma is challenging to clinically address. Although standard-of-care targeted therapy has high response rates in patients with BRAF-mutant melanoma, therapy relapse occurs in most cases. Intrinsically resistant melanoma cells drive therapy resistance and display molecular and biologic properties akin to neural crest-like stem cells (NCLSC) including high invasiveness, plasticity, and self-renewal capacity. The shared transcriptional programs and vulnerabilities between NCLSCs and cancer cells remains poorly understood. Here, we identify a developmental LPAR1-axis critical for NCLSC viability and melanoma cell survival. LPAR1 activity increased during progression and following acquisition of therapeutic resistance. Notably, genetic inhibition of LPAR1 potentiated BRAFi ± MEKi efficacy and ablated melanoma migration and invasion. Our data define LPAR1 as a new therapeutic target in melanoma and highlights the promise of dissecting stem cell-like pathways hijacked by tumor cells. SIGNIFICANCE: This study identifies an LPAR1-axis critical for melanoma invasion and intrinsic/acquired therapy resistance.

Tumor-infiltrating mast cells are associated with resistance to anti-PD-1 therapy.

Anti-PD-1 therapy is used as a front-line treatment for many cancers, but mechanistic insight into this therapy resistance is still lacking. Here we generate a humanized (Hu)-mouse melanoma model by injecting fetal liver-derived CD34+ cells and implanting autologous thymus in immune-deficient NOD-scid IL2Rγnull (NSG) mice. Reconstituted Hu-mice are challenged with HLA-matched melanomas and treated with anti-PD-1, which results in restricted tumor growth but not complete regression. Tumor RNA-seq, multiplexed imaging and immunohistology staining show high expression of chemokines, as well as recruitment of FOXP3+ Treg and mast cells, in selective tumor regions. Reduced HLA-class I expression and CD8+/Granz B+ T cells homeostasis are observed in tumor regions where FOXP3+ Treg and mast cells co-localize, with such features associated with resistance to anti-PD-1 treatment. Combining anti-PD-1 with sunitinib or imatinib results in the depletion of mast cells and complete regression of tumors. Our results thus implicate mast cell depletion for improving the efficacy of anti-PD-1 therapy.

Basal-Cell Carcinoma Occurring in Cutaneous Infundibular Cysts: Report of 2 Cases and Review of the Literature.

Cysts lined by stratified squamous epithelium indistinguishable from the epidermis, referred to as epidermoid cysts, epidermal inclusion cysts, and infundibular cysts, are the most common type of cyst occurring in the skin. They are invariably benign, and malignant neoplasms arising within the wall of such cysts are distinctly uncommon. Even basal-cell carcinoma, which is the most common cutaneous malignant neoplasm of the skin, has rarely been reported to occur in association with epidermoid cysts. The authors report their experience studying 2 patients with basal-cell carcinoma arising in association with an epidermoid cyst. These cases highlight the need to examine, histopathologically, tissue from this common and usually benign lesion. The authors also review the medical literature.

Education in skin cancer management--assessing knowledge and safety.

BACKGROUND: General practitioners manage the majority of skin cancers in Australia. There are a range of training opportunities for, and certifications in, skin cancer management. METHOD: Between 15 June and 25 June 2008, an online examination was placed on the Australasian College of Skin Cancer Medicine website. Two hundred and forty-five college affiliated doctors were invited by email to complete the examination. Thirty questions were asked pertaining to the management of a hypothetical case study including melanoma, basal cell carcinoma and squamous cell carcinoma. RESULTS: Of 187 doctors who had an active responding email address, 140 (75%) took the examination. From a possible score of 100, the mean score was 84 +/- 16. The median score was 80. DISCUSSION: Some trends emerged. Longer and more detailed training programs correlated with better subsequent knowledge retention and safety. Two days of training may not make doctors sufficiently safe in skin cancer management; it appeared to improved knowledge, but not to a point where unsafe practice was eliminated.

Family functioning and hope in children with juvenile rheumatoid arthritis.

PURPOSE: To examine the relationships among family functioning, hope, and quality of life in children with juvenile rheumatoid arthritis (JRA). STUDY DESIGN AND METHOD: Sixty-eight children (8 to 12 years of age) with a diagnosis of JRA and one of their parents/guardians participated in this descriptive correlational study. Parents completed the Feetham Family Functioning Survey (FFFS), the Parent Report for Children Pediatric Quality of Life Inventory (PedsQL), and the Parent Report for Children PedsQL Rheumatology Module. The children completed the Children's Hope Scale (CHS), the Child Report for PedsQL, and the Child Report PedsQL Rheumatology Module. Data were analyzed using chi-square, t-tests, and correlation analyses. RESULTS: Family functioning and children's hope showed a negative correlation, indicating that a child's hope was lower when the parent reported greater dissatisfaction with family functioning. Hope was not related to parent or child ratings of the child's quality of life. CLINICAL IMPLICATIONS: In caring for children with JRA, nurses can assess the family's satisfaction with relationships to the broader community, subsystems, and individual members and seek ways to promote healthy family functioning. Nurses also can assess the level of hope in children with JRA and facilitate the development of hopefulness by helping children establish goals and develop strategies to meet them.

A concept analysis of dignity for older adults.

BACKGROUND: Human dignity is an essential value of professional nursing education as well as a component of the American Nurses Association Code of Ethics. Nurses are exhorted to treat patients with dignity, and older adults want to be treated with dignity and die with dignity. Although dignity, particularly the dignity of older adults, is often discussed in the health care literature, its meaning is not always clear. AIM: The aim of this paper is to describe a concept analysis to develop a definition of dignity in older adults. METHODS: Data were collected using a literature review and five focus groups composed of older adults. The literature provided data about professionals' ideas of dignity and the focus groups provided qualitative data about the nature of dignity in older people. The literature review and focus groups were carried out concurrently, followed by synthesis of the findings. FINDINGS: Dignity is an inherent characteristic of being human, it can be subjectively felt as an attribute of the self, and is made manifest through behaviour that demonstrates respect for self and others. Dignity must be learned, and an individual's dignity is affected by the treatment received from others. CONCLUSIONS: A behavioural definition of dignity was constructed and this could provide the theoretical basis for nurses to develop interventions that foster dignity for older people.

Influence of indomethacin on lens regeneration in the newt notophthalmus viridescens.

Following lentectomy newts were injected with indomethacin in a variety of carrier solutions at doses ranging from 1.2-120 mg/kg body weight every other day for 15-17 days. The results show that injection of this drug according to the regimen used has no significant effect on regeneration of the lens. The data suggest, but do not prove, that prostaglandins may not play a major role in the early phases of lens regeneration in the newt.

A scanning electron microscopic study of differentiation of the digital pad in regenerating digits of the kenyan reed frog, Hyperolius viridiflavus ferniquei.

Newly metamorphosed Kenyan reed frogs, Hyperolius viridiflavus ferniquei, are able to regenerate amputated digits. The terminal digital pad is also completely reformed. Differentiation of the regenerating digital pad was studied by scanning electron microscopy. External differentiation of the digital pad began late in the second postamputational week with the appearance of small patches of specialized epidermal cells on the ventral surface of the regenerating digit. The differentiation of the pad spread out radially until late in the fourth week, when its overall shape approximated that of the normal digital pad. The appearance of patches of digital pad epidermis on the ends of spike regenerates arising from the forearm was also confirmed.