Food-Borne Biotoxin Neutralization in Vivo by Nanobodies: Current Status and Prospects.

Food-borne biotoxins from microbes, plants, or animals contaminate unclean, spoiled, and rotten foods, posing significant health risks. Neutralizing such toxins is vital for human health, especially after food poisoning. Nanobodies (Nbs), a type of single-domain antibodies derived from the genetic cloning of a variable domain of heavy chain antibodies (VHHs) in camels, offer unique advantages in toxin neutralization. Their small size, high stability, and precise binding enable effective neutralization. The use of Nbs in neutralizing food-borne biotoxins offers numerous benefits, and their genetic malleability allows tailored optimization for diverse toxins. As nanotechnology continues to evolve and improve, Nbs are poised to become increasingly efficient and safer tools for toxin neutralization, playing a pivotal role in safeguarding human health and environmental safety. This review not only highlights the efficacy of these agents in neutralizing toxins but also proposes innovative solutions to address their current challenges. It lays a solid foundation for their further development in this crucial field and propels their commercial application, thereby contributing significantly to advancements in this domain.

CAR-macrophage versus CAR-T for solid tumors: The race between a rising star and a superstar.

Adoptive cell therapy (ACT) has been demonstrated to be one of the most promising cancer immunotherapy strategies due to its active antitumor capabilities in vivo. Engineering T cells to overexpress chimeric antigen receptors (CARs), for example, has shown potent efficacy in the therapy of some hematologic malignancies. However, the efficacy of chimeric antigen receptor T cell (CAR-T) therapy against solid tumors is still limited due to the immunosuppressive tumor microenvironment (TME) of solid tumors, difficulty in infiltrating tumor sites, lack of tumor-specific antigens, antigen escape, and severe side effects. In contrast, macrophages expressing CARs (CAR-macrophages) have emerged as another promising candidate in immunotherapy, particularly for solid tumors. Now at its nascent stage (with only one clinical trial progressing), CAR-macrophage still shows inspiring potential advantages over CAR-T in treating solid tumors, including more abundant antitumor mechanisms and better infiltration into tumors. In this review, we discuss the relationships and differences between CAR-T and CAR-macrophage therapies in terms of their CAR structures, antitumor mechanisms, challenges faced in treating solid tumors, and insights gleaned from clinical trials and practice for solid tumors. We especially highlight the potential advantages of CAR-macrophage therapy over CAR-T for solid tumors. Understanding these relationships and differences provides new insight into possible optimization strategies of both these two therapies in solid tumor treatment.

Development of a Biotinylated Nanobody-Based Gold Nanoparticle Immunochromatographic Assay for the Detection of Procymidone in Crops.

A heavy-chain antibody (VHH) library against procymidone (PRM) was constructed via immunizing Bactrian camels. Through careful biopanning, seven nanobodies (Nbs) with different sequences were obtained. The variability in their performance was primarily attributed to the amino acid differences in complementarity-determining region 3 (CDR3), as analyzed by molecular docking. The Nb exhibiting the highest sensitivity, named NbFM5, was biotinylated and conjugated to streptavidin-labeled gold nanoparticles to preserve the epitope's activity and prevent a decrease in sensitivity due to traditional random electrostatic adsorption. Subsequently, a simple and sensitive immunochromatographic assay (ICA) was developed for rapid detection of PRM based on biotinylated Nb (btNb). The developed btNb-ICA showed a cut-off value of 200 ng/mL for visual judgment and a half-inhibitory concentration (IC50) of 6.04 ng/mL for quantitative detection. The limit of detection (LOD) was as low as 0.88 ng/mL. The recoveries in actual samples of crops ranged from 82.2 to 117.3%, aligning well with the results obtained from GC-MS/MS (R2 = 0.995). In summary, the developed btNb-ICA demonstrated high specificity and good accuracy for the rapid detection of PRM residues in vegetables. The total analysis time from preparing the sample to obtaining the result was less than 25 min.

A bispecific nanobody with high sensitivity/efficiency for simultaneous determination of carbaryl and its metabolite 1-naphthol in the soil and rice samples.

The simultaneous determination of carbaryl and its metabolite 1-naphthol is essential for risk assessment of pesticide exposure in agricultural and environmental samples. Herein, several bispecific nanobodies (BsNbs) with different lengths of hydrophilic linkers and junction sites were prepared and characterized for the simultaneous recognition of carbaryl and its metabolite 1-naphthol. It was found that the affinity of BsNbs to the analytes could be regulated by controlling linker length and linking terminal. Additionally, molecular simulation revealed that linker lengths affected the conformation of BsNbs, leading to alteration in sensitivity. The BsNb with G4S linker, named G4S-C-N-VHH, showing good thermal stability and sensitivity was used to develop a bispecific indirect competitive enzyme-linked immunosorbent assay (Bic-ELISA). The assay demonstrated a limit of detection of 0.8 ng/mL for carbaryl and 0.4 ng/mL for 1-naphthol in buffer system. Good recoveries from soil and rice samples were obtained, ranging from 80.0% to 112.7% (carbaryl) and 76.5%-110.8% (1-naphthol), respectively. Taken together, this study firstly provided a BsNb with high sensitivity and efficiency against environmental pesticide and its metabolite, and firstly used molecular dynamics simulation to explore the influence of linker on recognition. The results are valuable for the application of immunoassay with high efficiency in the fields of environment and agriculture.

Concurrent Waldenstrom's Macroglobulinemia and Myelodysplastic Syndrome with a Sequent t(10;13)(p13;q22) Translocation.

Myelodysplastic syndromes (MDS) and Waldenstrom's macroglobulinemia (WM) are rarely synchronous. Ineffective myelopoiesis/hematopoiesis with clonal unilineage or multilineage dysplasia and cytopenias characterize MDS. Despite a myeloid origin, MDS can sometimes lead to decreased production, abnormal apoptosis or dysmaturation of B cells, and the development of lymphoma. WM includes bone marrow involvement by lymphoplasmacytic lymphoma (LPL) secreting monoclonal immunoglobulin M (IgM) with somatic mutation (L265P) of myeloid differentiation primary response 88 gene (MYD88) in 80-90%, or various mutations of C-terminal domain of the C-X-C chemokine receptor type 4 (CXCR4) gene in 20-40% of cases. A unique, progressive case of concurrent MDS and WM with several somatic mutations (some unreported before) and a novel balanced reciprocal translocation between chromosomes 10 and 13 is presented below.

Refractory Splenic Marginal Zone Lymphoma Responsive to Combination Venetoclax and Bortezomib (Velcade) (V2) Therapy.

Standard treatment regimens for the management of patients with refractory splenic marginal zone lymphoma (SMZL) are currently unavailable. Here, we report a case of SMZL, which, after failing multiple therapeutics, demonstrated an impressive clinical response to combined Venetoclax and Velcade (V2), a treatment combination currently being investigated in the setting of refractory multiple myeloma. We also report a unique histopathology and mutational profile that may have important implications for the characterization and prognosis of SMZL.

Combined Ruxolitinib and Venetoclax Treatment in a Patient with a BCR-JAK2 Rearranged Myeloid Neoplasm.

Hematological malignancies with a BCR-JAK2 rearrangement have been described only sporadically in the literature over the last three decades. Although most patients suffer from a chronic myeloid neoplasm with marked eosinophilia, the clinical presentation varies significantly and can even manifest as a lymphoid malignancy. In this case report, we present a patient with a therapy-related BCR-JAK2 + myeloid neoplasm with extensive extramedullary disease localizing in the lymph nodes. While treatment with a JAK2 inhibitor (ruxolitinib) was not able to stop disease progression, combination treatment with inhibitors of both JAK2 and BCL2 (venetoclax) resulted in disease control for over 1.5 years. Combining these two inhibitors might be strategic in these patients, not only because BCL2 is a downstream target of JAK/STAT signaling but also because BCL2 is crucial for JAK2 inhibitor resistance. The recent inclusion of JAK2-rearranged malignancies in major classification systems and guidelines emphasizes the importance of not only getting a better understanding of the clinical phenotype of these rare disorders but also of identifying alternative treatment options for patients ineligible for allogeneic stem cell transplantation. Considering the low toxicity of combination treatment with these two small molecule inhibitors, this regimen could be further explored in future studies.

Hairy Cell Leukemia and Ground Water Contamination With Industrial Solvents: a Case Report.

The industrial solvents benzene and trichloroethylene (TCE) are known carcinogens, and these solvents contaminated the drinking water at Marine Corps Base Camp Lejeune from the 1950s to 1980s. Benzene and TCE are linked to the hematopoietic cancers acute myelocytic and lymphocytic leukemia and chronic lymphocytic leukemia. We report the case of a veteran stationed at Marine Corps Base Camp Lejeune during this period who developed hairy cell leukemia (HCL), a rare form of lymphocytic leukemia. We review his presentation, medical history, solvent exposure, and literature on the carcinogenicity of benzene and TCE. This patient represents a possible link of TCE or benzene to HCL. The case also informs clinicians of the updated epidemiology with regards to clinical findings for HCL.

Plasmablastic Lymphoma: Case Report of Prolonged Survival of an Advanced Human Immunodeficiency Patient and Literature Review.

Clinical Practice Points. Plasmablastic lymphoma (PBL) is a rare and highly aggressive variant of diffuse large B cell lymphoma with median survival of advanced stage patients varying between 6 and 15 months in previous reports. We report here a human immunodeficiency virus-infected patient surviving over 12 years following treatment for advanced PBL with EPOCH chemotherapy and intrathecal therapy. This case highlights the potential for improved survival in PBL with intensive chemotherapy. Further, literature review suggests promising prospects utilizing novel targeted therapies to increase the rate of prolonged responses.

TMSB4Y is a candidate tumor suppressor on the Y chromosome and is deleted in male breast cancer.

Male breast cancer comprises less than 1% of breast cancer diagnoses. Although estrogen exposure has been causally linked to the development of female breast cancers, the etiology of male breast cancer is unclear. Here, we show via fluorescence in situ hybridization (FISH) and droplet digital PCR (ddPCR) that the Y chromosome was clonally lost at a frequency of ~16% (5/31) in two independent cohorts of male breast cancer patients. We also show somatic loss of the Y chromosome gene TMSB4Y in a male breast tumor, confirming prior reports of loss at this locus in male breast cancers. To further understand the function of TMSB4Y, we created inducible cell lines of TMSB4Y in the female human breast epithelial cell line MCF-10A. Expression of TMSB4Y resulted in aberrant cellular morphology and reduced cell proliferation, with a corresponding reduction in the fraction of metaphase cells. We further show that TMSB4Y interacts directly with ß-actin, the main component of the actin cytoskeleton and a cell cycle modulator. Taken together, our results suggest that clonal loss of the Y chromosome may contribute to male breast carcinogenesis, and that the TMSB4Y gene has tumor suppressor properties.

Abdominal Wall Desmoid Fibromatosis Mimics Sarcoma With Intense FDG Uptake on FDG PET/CT.

We present the case of an abdominal wall soft tissue mass with intense FDG uptake in a 61-year-old man with unintentional weight loss. The appearance of this mass and high focal FDG uptake mimics high-grade sarcoma on (18)F-FDG PET/CT. However, the final excisional histopathological diagnosis was desmoid fibromatosis.

Breast cancer in male veteran population: an analysis from VA cancer registry.

BACKGROUND: Male breast cancer is rare and makes up < 1% of all cases of breast cancers. Treatment and survival stage per stage is mainly based on what is known from female breast cancer. OBJECTIVES: We determined the pathological features, stage, treatment of breast cancer in male veterans and their survival outcome. METHODS: Medical records of male patients diagnosed with breast cancer at the Veterans Affairs Medical Centers of Washington DC, Baltimore, Maryland, and Martinsburg, West Virginia, from 1992-2012 were reviewed after iInstitutional review board approval. RESULTS: From 1995-2012, 51 male patients with breast cancer were identifed from cancer registry. Of those, 57% were African American, 41% white, and 2% other race. Median age was 68 years (range, 44-86 years). Palpable mass was presenting symptoms in 80%, and gynecomastia or bloody nipple discharge in 16%. Family history of breast cancer in immediate family was positive in 11 patients without mention of BRCA genes except in one who was BRCA2-positve. ER/PR (estrogen-/progesterone-receptor) was positive in 71%, ER-positive/PR-negative in 2%, ER-positive/PR-positive /HER2-positive in 4%, ER-negative/PR-negative /HER2-triple negative in 4%. In all, 41% and 57% had right and left breast cancer, respectively; 80% had mastectomy, 36% had lymph node involvement (1-13 LN), 90% had invasive ductal carcinoma, 8% DCIS, and 2% sarcoma. Cancer in 26% was stage I, 38% stage II, 18% stage III and 8% stage IV. Twenty four percent of the patients had combination chemotherapy, and 66% were given tamoxifen. Eight percent had relapsed or recurrent disease within 1-5 years of their diagnosis and died within 2-12 years after the relapse. At median follow-up of 174 months (range, 4 months-19 years), 56% had died, 42% were alive, and 6% had been lost to follow-up. LIMITATIONS: This is a very small retrospective chart review. Further large prospective studies are desired. CONCLUSIONS: Median age at diagnosis of breast cancer seems to be higher in men (70 years) than it is in women (60 years). Invasive ductal carcinoma is the main pathology, and 73% of the tumors were ER-positive. The survival rate at more than 10 years of follow-up was about 40%. Stage versus survival revealed no difference in mortality.

Tumor-infiltrating immune cells: triggers for tumor capsule disruption and tumor progression?

BACKGROUND: Our previous studies of human breast and prostate cancer have shown that aberrant immune cell infiltration is associated with focal tumor capsule disruption and tumor cell budding that facilitate invasion and metastasis. Our current study attempted to determine whether aberrant immune cell infiltration would have similar impact on colorectal cancer (CRC). MATERIALS AND METHODS: Tissue sections from 100 patients with primary CRC were assessed for the frequencies of focal basement membrane (BM) disruption, muscularis mucosa (MM) fragmentation, and tumor cell dissemination in epithelial structures adjacent and distal to infiltrating lymphoid aggregates using a panel of biomarkers and quantitative digital imaging. RESULTS: Our study revealed: (1) epithelial structures adjacent to lymphoid follicles or aggregates had a significantly higher (p<0.001) frequency of focally disrupted BM, dissociated epithelial cells in the stroma, disseminated epithelial cells within lymphatic ducts or blood vessels, and fragmented MM than their distal counterparts, (2) a majority of dissociated epithelial cells within the stroma or vascular structures were immediately subjacent to or physically associated with infiltrating immune cells, (3) the junctions of pre-invasive and invasive lesions were almost exclusively located at sites adjacent to lymphoid follicles or aggregates, (4) infiltrating immune cells were preferentially associated with epithelial capsules that show distinct degenerative alterations, and (5) infiltrating immune cells appeared to facilitate tumor stem cell proliferation, budding, and dissemination. CONCLUSIONS: Aberrant immune cell infiltration may have the same destructive impact on the capsule of all epithelium-derived tumors. This, in turn, may selectively favor the proliferation of tumor stem or progenitor cells overlying these focal disruptions. These proliferating epithelial tumor cells subsequently disseminate from the focal disruption leading to tumor invasion and metastasis.

Concurrent diagnoses of Hodgkin lymphoma and biclonal myeloma in the bone marrow.

The diagnoses of Hodgkin lymphoma and multiple myeloma have rarely been made simultaneously in the same patient. We present a case of an 82-year-old man who rapidly developed pancytopenia and liver failure with coagulopathy. Serum protein electrophoresis and immunofixation revealed an unequivocal immunoglobulin Gkappa and immunoglobulin Glambda biclonal gammopathy. Bone marrow biopsy showed involvement by classic Hodgkin lymphoma with an inflammatory background including 49% mature plasma cells. Unfortunately, the patient died 14 days after admission. To our knowledge, a case of concurrent Hodgkin lymphoma and biclonal multiple myeloma has not previously been reported. Detection of severe bone marrow plasmacytosis in the background of Hodgkin lymphoma should alert the pathologist to the possibility of collision with a plasma cell neoplasm, warranting a complete diagnostic workup.

CD4/CD8 double negative pagetoid reticulosis: a case report and literature review.

Pagetoid reticulosis is an indolent primary cutaneous T-cell lymphoma. It typically presents as a solitary and slowly growing patch or plaque on the extremity, histologically characterized by an acanthotic epidermis infiltrated with atypical lymphocytes. Here, we present histological, immunophenotypical and molecular findings of a 29-year-old Jamaican man with bilateral wrist plaques. Histology showed marked acanthosis, hyperkeratosis and an intraepidermal infiltration consisting of large atypical lymphocytes. Immunohistochemical stains showed CD3 and CD5 positive T cells with significant loss of CD7, double negative CD4 and CD8 and strong positive CD30. Molecular analysis showed a monoclonal T-cell receptor (TCR) gamma gene rearrangement. Review of the literature confirms that the immunophenotype of pagetoid reticulosis is variable with decreasing frequency of CD8+ cytotoxic/suppressor T cell, CD4+ helper T cell and least commonly CD4/CD8 double negative phenotypes. Although CD4/CD8 double negative phenotype appears to be associated with higher proliferation index, it does not appear to confer prognostic significance.