More than Just Skin-Deep: A Review of Imaging's Role in Guiding CAR T-Cell Therapy for Advanced Melanoma.

Advanced melanoma is one of the deadliest cancers, owing to its invasiveness and its propensity to develop resistance to therapy. Surgery remains the first-line treatment for early-stage tumors but is often not an option for advanced-stage melanoma. Chemotherapy carries a poor prognosis, and despite advances in targeted therapy, the cancer can develop resistance. CAR T-cell therapy has demonstrated great success against hematological cancers, and clinical trials are deploying it against advanced melanoma. Though melanoma remains a challenging disease to treat, radiology will play an increasing role in monitoring both the CAR T-cells and response to therapy. We review the current imaging techniques for advanced melanoma, as well as novel PET tracers and radiomics, in order to guide CAR T-cell therapy and manage potential adverse events.

The effects of selected asanas in Iyengar yoga on flexibility: pilot study.

In recent years the practice of yoga has gained popularity as a form of physical fitness and exercise, and has been said to improve strength and flexibility. The main objective of this research project was to evaluate the effects of a six week Iyengar yoga intervention on flexibility. N = 16 low to moderately active females (52.37 ± 7.79 years) attended Iyengar yoga practice for a total of 6 weeks, consisting of one 90 min session per week. Lumbar and hamstring flexibility were assessed pre and post-intervention using a standard sit and reach test. The results show a significant increase in flexibility, indicating 6 weeks of single session yoga training may be effective in increasing erector spinae and hamstring flexibility. This is important when considering that much of the population find it difficult to attend more than one session a week into their training schedule.

The relationship between ankle joint physiological characteristics and balance control during unilateral stance.

BACKGROUND: The role that the ankle's physiological characteristics play in maintaining balance during quiet stance has been well documented. However, the role of the ankle in maintaining balance during more challenging conditions is questionable. As such, the objectives of this study were to identify any significant relationships between the physiological characteristics of the ankle joint and the ability to maintain more challenging unilateral stance. PARTICIPANTS: 21 healthy, adult athletes (age = 24.67 ± 5.42 years; height = 175.34 ± 7.48 cms; weight = 79.09 ± 14.07 kg). PROCEDURES: Passive resistance and joint position sense in the sagittal plane of the ankle, and active dorsiflexion range of motion of each subject was assessed, in addition to centre of pressure parameters during 20 s unilateral stance. RESULTS: Pearson's product moment correlation coefficient found significant positive correlations between D(peaktorque) and sway area (r = .554); A(x) range (r = .449); and A(y) range (r = .471). Significant negative correlations were found between P(peaktorque) angle and sway area (r = -.538, p = .012), A(x) range (r = -.590, p = .005) and A(y) range (r = -.439, p=.046). DISCUSSION: The results highlighted limited relationships between unilateral stance balance control and the ankle characteristics commonly associated with quiet stance balance control and has, thus, further questioned the role that the ankle plays during more challenging stance conditions. The majority of balance training protocols in the athletic community focuses on the distal joints, however, this needs re-addressing in order to maximise performance.

Distinct Toll-like receptor signals regulate cerebral parasite load and interferon α/ß and tumor necrosis factor α-dependent T-cell infiltration in the brains of Trypanosoma brucei-infected mice.

BACKGROUND: The penetration of T cells and trypanosomes into the brain parenchyma is a major pathogenetic event in African trypanosomiasis. METHODS: The role of innate immune responses in the penetration of T cells and Trypanosoma brucei brucei into the brain was studied in knockout mice by using double immunofluorescent staining and real-time polymerase chain reaction. RESULTS: We demonstrate that Toll-like receptor (TLR)-MyD88-mediated signaling is required for T-cell and parasite penetration into the brain and microglial activation, besides controlling parasitemia and antigen-specific T-cell activation. Among different TLR-deficient mice studied, TLR9 mediated parasitemia control and T-cell penetration into the brain. TLR-MyD88 signals increased levels of interferon (IFN) ß and tumor necrosis factor (TNF) α transcripts in the brains of infected mice and both TNF-α and IFN-α/ß, receptors promoted T-cell and trypanosoma infiltration into the brain parenchyma. Both resident and infiltrating inflammatory cells in the brain controlled parasite densities in a TLR2- and TLR9-MyD88-mediated manner. However, neither IFN-α/ß nor TNF-α contributed to parasite control in the brain. CONCLUSIONS: Our data indicate that innate immune TLR signals stimulate the expression of TNF-α and IFN-α/ß that initiate brain invasion of T cells and trypanosomes, and control T. brucei brucei load in the brain by molecules distinct from these.

Identification of stage biomarkers for human African trypanosomiasis.

Human African trypanosomiasis (HAT), caused by infection with sub-species of Trypanosoma brucei (T. b.), manifests as a hemolymphatic stage followed by an encephalitic stage. The distinction of the two stages needs improvement as drugs used for the late stage are highly toxic. Transcripts encoding 16 secreted proteins differentially expressed in the brains of mice at late stage T. b. brucei infection when the early stage drug suramin is no longer effective and different to immunoglobulins, chemokines, and cytokines, were selected by microarray analysis. Lipocalin 2 and secretory leukocyte peptidase inhibitor (SLPI) mRNA showed the highest differential expression in mice. These transcripts were also upregulated in brains from infected rats. Lipocalin 2 was increased in cerebrospinal fluid (CSF) from rats during late stage T. b. brucei infection. Protein levels of lipocalin 2, SLPI, and the chemokine CXCL10 were found increased in CSF from Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense late stage HAT compared to early stage.

Expression and role of CXCL10 during the encephalitic stage of experimental and clinical African trypanosomiasis.

BACKGROUND: Human African trypanosomiasis, caused by Trypanosoma brucei, involves an early hemolymphatic stage followed by a late encephalitic stage. METHODS: We studied the expression of chemokines with use of microarray and enzyme-linked immunosorbent assay in T. brucei brucei-infected mice and in patients with human African trypanosomiasis and examined their role in controlling brain accumulation of T cells and parasites. RESULTS: The messenger RNAs (mRNAs) encoding CXCR3 ligands CXCL9 and CXCL10 demonstrated the greatest increases among chemokines in brain specimens of infected mice, as determined by microarray. CXCL9 and CXCL10 mRNA accumulation was interferon (IFN)-gamma-dependent. Expression of CXCL10 was predominantly observed in astrocytes. Weight loss was registered in wild-type but not in CXCL10(-/-) and CXCR3(-/-) infected mice. Infected CXCL10(-/-) or CXCR3(-/-) mice demonstrated reduced accumulation of trypanosomes and T cells in the brain parenchyma but similar parasitemia levels, compared with wild-type mice. CXCL10 and IFN-gamma levels were increased in the cerebrospinal fluid of patients with late stage but not early stage human African trypanosomiasis. Levels of CXCL10 in patients with late stage human African trypanosomiasis were associated with somnolence, low body weight, and trypanosomes in the cerebrospinal fluid. CONCLUSION: IFN-gamma-dependent CXCL10 is critical for accumulation of T cells and trypanosomes in the brain during experimental African trypanosomiasis. Data suggest CXCL10 as a candidate marker for late stage human African trypanosomiasis.

Preclinical assessment of the treatment of second-stage African trypanosomiasis with cordycepin and deoxycoformycin.

BACKGROUND: There is an urgent need to substitute the highly toxic compounds still in use for treatment of the encephalitic stage of human African trypanosomiasis (HAT). We here assessed the treatment with the doublet cordycepin and the deaminase inhibitor deoxycoformycin for this stage of infection with Trypanosoma brucei (T.b.). METHODOLOGY/PRINCIPAL FINDINGS: Cordycepin was selected as the most efficient drug from a direct parasite viability screening of a compound library of nucleoside analogues. The minimal number of doses and concentrations of the drugs effective for treatment of T.b. brucei infections in mice were determined. Oral, intraperitoneal or subcutaneous administrations of the compounds were successful for treatment. The doublet was effective for treatment of late stage experimental infections with human pathogenic T.b. rhodesiense and T.b. gambiense isolates. Late stage infection treatment diminished the levels of inflammatory cytokines in brains of infected mice. Incubation with cordycepin resulted in programmed cell death followed by secondary necrosis of the parasites. T.b. brucei strains developed resistance to cordycepin after culture with increasing concentrations of the compound. However, cordycepin-resistant parasites showed diminished virulence and were not cross-resistant to other drugs used for treatment of HAT, i.e. pentamidine, suramin and melarsoprol. Although resistant parasites were mutated in the gene coding for P2 nucleoside adenosine transporter, P2 knockout trypanosomes showed no altered resistance to cordycepin, indicating that absence of the P2 transporter is not sufficient to render the trypanosomes resistant to the drug. CONCLUSIONS/SIGNIFICANCE: Altogether, our data strongly support testing of treatment with a combination of cordycepin and deoxycoformycin as an alternative for treatment of second-stage and/or melarsoprol-resistant HAT.

Suramin and minocycline treatment of experimental African trypanososmiasis at an early stage of parasite brain invasion.

The effect of treatment on relapses of Trypanosoma brucei (T. b.) brucei infections in mice in relation to passage of the parasites across the blood-brain barrier (BBB) as visualized by immunohistochemistry was studied. Three daily intraperitoneal injections of 20mg/kg suramin starting at 15 days post-infection (p.i.), when trypanosomes had begun to traverse the BBB, were curative, but not when starting at 21 days p.i. when parasite brain invasion was more pronounced. Relapses occurred in all mice after one or two daily injections of suramin starting at 15 days p.i., but they were delayed when treatment was supplemented with minocycline, which impedes penetration of T. b. brucei into the brain. This study supports the notion that suramin may be effective even when minor parasite neuroinvasion has appeared in African trypanosomiasis and it shows that minocyline can affect relapses of the disease.

Glossina proteolytic lectin does not require a carbohydrate moiety for enzymatic or trypanosome-transforming activities.

The developmental cycle of the cyclically transmitted African trypanosome involves an obligatory passage through the tsetse fly, Glossina spp. This intricate relationship requires the presence of molecules within the insect vector, including a midgut lectin, that interact with the trypanosome. Recently, a gene encoding for a proteolytic lectin, with trypanosome-transforming activity, was isolated from a midgut cDNA library of Glossina fuscipes fuscipes Austen in our laboratory. Using the same approach, we have identified a similar gene from a midgut cDNA library of Glossina austeni (Newstead). The protein encoded by this gene was expressed in bacteria and a baculovirus-based expression system. The baculovirus-expressed lectin was found in the medium of baculovirus-infected Sf-21 cell cultures, indicating that the tsetse fly-derived signal peptide was recognized and cleaved by the Sf-21 cells. The baculovirus-expressed protein also was glycosylated despite the absence of classical O-linked and N-linked sugar attachment motifs. Both the baculovirus- and bacterium-expressed lectin proteins were shown to agglutinate trypanosomes and rabbit red blood cells in vitro. This agglutination was strongly inhibited by D-glucosamine. D-Glucosamine also inhibited the action of the authentic and recombinant lectins upon the chromogenic substrate Chromozym TRY. Interestingly, both baculovirus- and bacterium-expressed lectins showed no significant differences in terms of these activities, indicating that a sugar moiety is not essential for biological activity. Our results provide an important molecular tool for further characterization of Glossina proteolytic lectin.