Shawan Karan

Males can influence future mating interactions of females after copulation by changing female signals that subsequent males will encounter. In insects, such effects commonly involve cuticular hydrocarbons (CHCs), but whether heritable microbial symbionts contribute to post-mating chemical signalling remains largely unknown. Kelly’s citrus thrips, Pezothrips kellyanus , provides an ideal system to address this question because reproductive compatibility is shaped by common arthropod endosymbionts. Across P. kellyanus populations, Cardinium occurs in almost all individuals whereas Wolbachia varies in prevalence and appears to spread by cytoplasmic incompatibility (CI). Yet, females with only Cardinium (C) avoid incompatible males carrying both Cardinium and Wolbachia (CW), and this discrimination is linked to the distinct CHC profile of CW males. Here, we tested whether this endosymbiont-associated male perfume persists on females beyond copulation by altering the female CHC profile and subsequent male mating behaviour. Using behavioural assays and GC–MS-based CHC profiling, we found that, independent of female endosymbiont association, females first mated with CW males received fewer antennal contacts and mating attempts from subsequent C males. Furthermore, mating remodelled female CHC profiles, while mating with CW males produced a distinctive post-mating chemical signature. Most notably, tridecane, previously detected only in CW males, occurred exclusively in females mated with CW males. Our findings show that endosymbionts can alter mated female CHCs and influence future sexual communication between male and female hosts. These findings reveal a previously unrecognised post-mating route through which endosymbionts reshape sexual communication, with potential consequences for reproductive compatibility and symbiont transmission dynamics.

Fluorouracil‐platinum(IV) prodrugs represent a novel class of multimechanistic chemotherapeutics with enhanced anticancer potential. The prodrugs Pt ^IV P‐5FUMeOBut and Pt ^IV 56‐5FUMeOBut were actualized by derivatising the clinical drug 5‐fluorouracil (5FU) and coordinating it to platinum(IV) complexes, leveraging the established cytotoxicity of their platinum(II) precursors, Pt ^II PHEN SS and Pt ^II 56ME SS . The rationale behind this design is to combine the DNA synthesis‐disrupting antimetabolite activity of 5FU with platinum‐based cytotoxicity, aiming to enhance therapeutic efficacy. This study evaluates the in vitro potency of Pt ^IV P‐5FUMeOBut and Pt ^IV 56‐5FUMeOBut in colorectal cancer models, assessing their cellular uptake, cytotoxicity, and effects in 3D spheroid cultures. Comparative analyses with cisplatin and 5FU explore their impact on cell survival, reactive oxygen species production, mitochondrial membrane potential, and cell‐cycle progression. Mechanistic insights are further examined through apoptosis and necrosis assays, proteomic profiling, and western blot analysis of key signalling pathways. These findings contribute to the preclinical development of fluorouracil‐platinum(IV) prodrugs as potential candidates for cancer therapy.

Maternally transmitted endosymbionts of arthropods are common and phylogenetically diverse. Several bacteria, including Wolbachia and Cardinium, have independently evolved the ability to induce cytoplasmic incompatibility (CI) limiting the reproduction in females lacking the endosymbionts carried by their mates. While promoting endosymbiont spread, CI is costly to endosymbiont-free females. Such host-endosymbiont conflicts are expected to affect host mating preferences, yet this has scarcely been studied in hosts carrying multiple, potentially competing, endosymbionts. We investigated mate choice and chemical communication in a significant pest of citrus, Kelly’s citrus thrips (Pezothrips kellyanus), naturally carrying CI-inducing Cardinium and Wolbachia. Unlike females with both endosymbionts (CW) that had no preference for males with particular endosymbiont associations, females with only Cardinium (C) preferred compatible C and endosymbiont-free males over incompatible CW males. In contrast, endosymbiont-free females showed no preference, despite experiencing similar CI risks when facing incompatible C and CW males. Male mating success, however, mostly depended on female receptivity and not on endosymbiont association. Furthermore, chemical analyses revealed that males with different endosymbiont associations had distinctly different cuticular hydrocarbon (CHC) profiles, with the CHC profile of CW males markedly including tridecane, a compound known to influence animal behavior. The results indicate that Cardinium enables females to avoid Wolbachia-induced CI based on the distinct chemical cues of incompatible males. Our findings highlight the role of common endosymbionts and their interactions in sexual selection through their effects on chemical and behavioral traits of hosts, emphasizing the importance of these factors in endosymbiont and host population dynamics, as well as endosymbiont-based pest control strategies.

Chlorambucil-platinum(IV) prodrugs exhibit multi-mechanistic chemotherapeutic activity with promising anticancer potential. The platinum(II) precursors of the prodrugs have been previously found to induce changes in the microtubule cytoskeleton, specifically actin and tubulin of HT29 colon cells, while chlorambucil alkylates the DNA. These prodrugs demonstrate significant anticancer activity in 2D cell and 3D spheroid viability assays. A notable production of reactive oxygen species has been observed in HT29 cells 72 h post treatment with prodrugs of this type, while the mitochondrial membrane potential was substantially reduced. The cellular uptake of the chlorambucil-platinum(IV) prodrugs, assessed by ICP-MS, confirmed that active transport was the primary uptake mechanism, with platinum localisation identified primarily in the cytoskeletal fraction. Apoptosis and necrosis were observed at 72 h of treatment as demonstrated by Annexin V-FITC/PI assay using flow cytometry. Immunofluorescence measured via confocal microscopy showed significant changes in actin and tubulin intensity and in architecture. Western blot analysis of intrinsic and extrinsic pathway apoptotic markers, microtubule cytoskeleton markers, cell proliferation markers, as well as autophagy markers were studied post 72 h of treatment. The proteomic profile was also studied with a total of 1859 HT29 proteins quantified by mass spectroscopy, with several dysregulated proteins. Network analysis revealed dysregulation in transcription, MAPK markers, microtubule-associated proteins and mitochondrial transport dysfunction. This study confirms that chlorambucil-platinum(IV) prodrugs are candidates with promising anticancer potential that act as multi-mechanistic chemotherapeutics.

Development of resistance to cisplatin, oxaliplatin and carboplatin remains a challenge for their use as chemotherapies, particularly in breast and colorectal cancer. Here, we compare the anticancer effect of novel complexes [Pt(1,10-phenanthroline)(1S,2S-diaminocyclohexane)](NO₃)₂ (Pt^IIPHENSS), [Pt(5-methyl-1,10-phenanthroline)(1S,2S-diaminocyclohexane)](NO₃)₂ (Pt^II5MESS) and [Pt(5,6-dimethyl-1,10-phenanthroline)(1S,2S-diaminocyclohexane)](NO₃)₂ (Pt^II56MESS) and their platinum(IV)-dihydroxy derivatives with cisplatin. Complexes are greater than 11-fold more potent than cisplatin in both 2D and 3D cell line cultures with increased selectivity for cancer cells over genetically stable cells. ICP-MS studies showed cellular uptake occurred through an active transport mechanism with considerably altered platinum concentrations found in the cytoskeleton across all complexes after 24 h. Significant reactive oxygen species generation was observed, with reduced mitochondrial membrane potential at 72 h of treatment. Late apoptosis/necrosis was shown by Annexin V-FITC/PI flow cytometry assay, accompanied by increased sub-G0/G1 cells compared with untreated cells. An increase in S and G2+M cells was seen with all complexes. Treatment resulted in significant changes in actin and tubulin staining. Intrinsic and extrinsic apoptosis markers, MAPK/ERK and PI3K/AKT activation markers, together with autophagy markers showed significant activation of these pathways by Western blot. The proteomic profile investigated post-72 h of treatment identified 1597 MDA−MB−231 and 1859 HT29 proteins quantified by mass spectroscopy, with several differentially expressed proteins relative to no treatment. GO enrichment analysis revealed a statistically significant enrichment of RNA/DNA-associated proteins in both the cell lines and specific additional processes for individual drugs. This study shows that these novel agents function as multi-mechanistic chemotherapeutics, offering promising anticancer potential, and thereby supporting further research into their application as cancer therapeutics.

Four platinum(IV) prodrugs incorporating a biotin moiety to selectively target cancer cells were synthesised, characterised, and their biological activity assessed. All complexes exhibited exceptional in vitro cytotoxicity against a panel of cancer cell lines, with [Pt(5,6-dimethyl-1,10-phenanthroline)(1S,2S-diaminocyclohexane)(biotin)(hydroxido)](NO₃)₂, (2) exhibiting the lowest GI₅₀ of 4 nM in the prostate Du145 cancer cell line. Each complex displayed significantly enhanced activity compared to cisplatin, with 2 being 1000-fold more active in the HT29 colon cancer cell line. Against the MCF-7 breast cancer cell line, in which high levels of biotin receptors are expressed, 2, [Pt(4,7-dimethoxy-1,10-phenanthroline)(1S,2S-diaminocyclohexane)(biotin)(hydroxido)](NO₃)₂, (3), and [Pt(5-methyl-1,10-phenanthroline)(1S,2S-diaminocyclohexane)(biotin)(hydroxido)](NO₃)₂, (4) exhibited enhanced activity compared to their platinum(II) cores, with 4 being 6-fold more active than its platinum(II) precursor. Furthermore, 3 exhibited 3-fold greater selectivity towards MCF-7 breast cancer cells compared to MCF10A breast healthy cells, and this was further confirmed by platinum uptake studies, which showed 3 to have almost 3-fold greater uptake in MCF-7 cells, compared to MCF10A cells. The results show that lipophilicity and selectivity both contributed to the cellular uptake of 1–4; however, this was not always translated to the observed cytotoxicity.

Platinum(IV) prodrugs of the [Pt(P_L)(A_L)(COXi)(OH)]²⁺ type scaffold (where P_L is 1,10-phenanthroline or 5,6-dimethyl-1,10-phenanthroline, A_L is 1S,2S-diaminocyclohexane, and COXi is a COX inhibitor, either indomethacin or aspirin) were synthesised and characterised, and their biological activity was explored. MTT assays showed that these complexes exhibit outstanding activity against a range of cancer cell lines, and nanomolar activities were observed. The most potent complex, 4, exhibited a GI₅₀ of 3 nM in the Du145 prostate cancer cell line and was observed to display a 1614-fold increased activity against the HT29 colon cancer cell line relative to cisplatin. ICP-MS studies showed a linear correlation between increased cellular accumulation of the complexes and increased cytotoxicity, while an enzyme immunoassay showed that 1 and 2 inhibited COX-2 at 14 and 1.4 µM, respectively, which is comparable to the inhibition exhibited by indomethacin. These results suggest that while the cytotoxicity of prodrugs 1–4 was influenced by cellular uptake, it was not entirely dependent on either COX inhibition or lipophilicity.