Silencing of the Hsf gene, the transcriptional regulator of A. gambiae male accessory glands, inhibits the formation of the mating plug in mated females and disrupts their monogamous behaviour

Discovering the molecular factors that shape the mating behaviour and the fertility of the mosquito Anopheles gambiae, the principal vector of human malaria, is regarded as critical to better understand its reproductive success as well as for identifying new leads for malaria control measures. In A. gambiae mating induces complex behavioural and physiological changes in the females, including refractoriness to subsequent mating and induction of egg-laying. In other insects including Drosophila a group of proteins named Accessory gland proteins (Acps), produced by males and transferred with sperm to the female reproductive tract, have been implicated in this post-mating response. Although Acps represent a set of promising candidates for unravelling the mating physiology, their role in inducing behavioural changes in mated A. gambiae females remains largely unknown. In this work, we demonstrate that a down-regulation of a large fraction of Acp genes via silencing of the Acp regulating transcription factor Hsf, abolishes the formation of mating plug in mated females and fails to induce refractoriness of mated female to subsequent inseminations. A significant fraction of females mated to Hsf silenced males (66%) failed to receive the mating plug though seminal fluid had been transferred as documented by the presence of spermatozoa in the female sperm storage organ. Furthermore, nearly all females (95%) mated to HSF-silenced males were re-inseminated when exposed to males carrying EGPF marked sperm. Our findings provide evidence showing that Acp genes regulated by the transcription factor HSF play a key role in the function of the male accessory glands

Eccessiva durata del processo e responsabilità disciplinare dei magistrati : il ritardo nel deposito dei provvedimenti fra storia e attualità

ll tema dell’eccessiva durata dei processi e delle sue conseguenze pregiudizievoli sull’effettività della tutela giudiziaria rappresenta, da sempre, uno dei principali nodi del rapporto tra giustizia e opinione pubblica. Sin dall’unificazione italiana, la questione è stata affrontata per lo più sul piano delle riforme del processo e dell’ordinamento giudiziario, senza, tuttavia, la predisposizione di adeguati interventi in grado di incidere sull’organizzazione delle strutture e del personale. Se soltanto di recente il nostro ordinamento ha approntato strumenti di tutela diretta al principio della ragionevole durata del processo, formalizzato nell’art. 111 Cost., la responsabilità disciplinare dei magistrati per ritardo nel deposito dei provvedimenti ha rappresentato, sin dall’inizio del Novecento, uno degli strumenti principali non solo per reprimere gli episodi più gravi (oggi determinanti addirittura un danno erariale da disservizio), ma anche per restituire credibilità alla funzione giudiziaria, nell’ambito delle complesse dinamiche relative al rapporto tra magistratura e società. Lo studio mira a valutare se, in assenza di idonei strumenti normativi volti ad evitare o quantomeno a contenere il fenomeno della lunghezza dei procedimenti, gli interventi della giurisprudenza, prima, della Suprema corte disciplinare e, dopo, della Sezione disciplinare del Consiglio superiore della magistratura, chiamate a sanzionare gli illeciti dei singoli magistrati, siano stati in grado, nella difficoltà di trovare un equilibrio tra standard di rendimento e carichi esigibili, di rispondere in maniera soddisfacente al contenimento dei tempi del processo e alla riduzione dell'arretrato, obiettivi tra i principali del PNRR.The matter on the excessive duration of the judicial processes and its damaging consequences about the effects of the legal protection has always been one of the main issues in the relationship between justice and public opinion. Since the Italian unification, the problem has been mainly considered from the point of view of both judicial process reforms and judicial system reforms, without, however, providing adequate means to affect the organization of the structure and the personnel. It is just a short time since our legal system provided protection resources based on the principle of a fair duration of the judicial process, established in Article 111 of the Constitution. Whereas, since the beginning of the twentieth century, the disciplinary liability of judges, due to the delay in filing judicial orders, has represented one of the main resource not only to contain the most serious events (nowadays they cause even fiscal damage by disservice) but also to restore credibility in the judicial activity for the complicated dynamics related to the relationship between judiciary and society. Both the matters on the disciplinary liability of judges and the judiciary independence are interwoven in Italian history after unification and they had to find a delicate equilibrium. If during the liberal age and the Fascism, the disciplinary liability – according to the nineteenth-century model of the judge as an officer integrated in the state apparatus – allowed the control over judges – which was largely guaranteed, directly or not, by the executive power –, in republican Italy, instead, it fulfilled the aim to ensure the regular execution of the judicial activity thanks to a self-government system and to a different judge’ role inside the judicial system. The disciplinary bodies that were called to evaluate the judges’ conduct – the Supreme Disciplinary Court before and then after the year 1958 the Disciplinary Division of the Superior Council of the Judiciary – have realized a balance between the requests for reducing the long time frames in justice and the need to protect the prestige of judges. Referring to the case law of the Disciplinary Division of the Superior Council of the Judiciary, the Castelli reform introduced a standardization of disciplinary wrong, including the delay in filing judicial orders. It is regulated by art. 2 paragraph 1 letter q, of Legislative Decree of 23 february 2006 no. 109, and in accordance with it, the «repeated, serious and unjustified delay to carry out the job in the exercise of their duties» constitutes a wrong; «it is supposed that a delay is not serious when it does not exceed three times the period provided for by the law to carry out the job, except it may be proved otherwise». Such prediction represents already an improvement over the last years, where the disciplinary judge – in the absence of a rule considering the conduct of the ‘idle’ judge as a particular illegal case – could punish the most striking cases in a discretionary and variable manner, and not without contradiction. So these cases showed a clear violation by the judge who was blamed for a lack of diligence and his negligence was often accompanied by a complaint from the lawyers. At present in accordance with the provisions, three different and concurrent conditions must exist for the delay to be illegal: reiteration, seriousness of the delay and a lack of justification. These conditions (that are here studied according to the case law) are therefore all essential for the punishment of the delays. This study aims to determine if the case law measures, in the absence of suitable regulatory means for avoiding or at least containing the phenomenon of the slowness of proceedings, firstly, by the Supreme Disciplinary Court and, later, by the Disciplinary Division of the Superior Council of the Judiciary which had to punish the wrong of each judge, have been able to find a difficult balance between the standards of performance and a bearable workload. Furthermore if they were able to contain satisfactorily both the judicial process time and the delays, which are some of the main goals of the National Recovery Plan and Resilience

Sulforaphane-loaded ultradeformable vesicles as a potential natural nanomedicine for the treatment of skin cancer diseases

Sulforaphane is a multi-action drug and its anticancer activity is the reason for the continuous growth of attention being paid to this drug. Sulforaphane shows an in vitro antiproliferative activity against melanoma and other skin cancer diseases. Unfortunately, this natural compound cannot be applied in free form on the skin due to its poor percutaneous permeation determined by its physico-chemical characteristics. The aim of this investigation was to evaluate ethosomes® and transfersomes® as ultradeformable vesicular carriers for the percutaneous delivery of sulforaphane to be used for the treatment of skin cancer diseases. The physico-chemical features of the ultradeformable vesicles were evaluated. Namely, ethosomes® and transfersomes® had mean sizes <400 nm and a polydispersity index close to 0. The stability studies demonstrated that the most suitable ultradeformable vesicles to be used as topical carriers of sulforaphane were ethosomes® made up of ethanol 40% (w/v) and phospholipon 90G 2% (w/v). In particular, in vitro studies of percutaneous permeation through human stratum corneum and epidermis membranes showed an increase of the percutaneous permeation of sulforaphane. The antiproliferative activity of sulforaphane-loaded ethosomes® was tested on SK-MEL 28 and improved anticancer activity was observed in comparison with the free drug

Hitting the right note at the right time: Circadian control of audibility in Anopheles mosquito mating swarms is mediated by flight tones

Mating swarms of malaria mosquitoes form every day at sunset throughout the tropical world. They typically last less than 30 minutes. Activity must thus be highly synchronized between the sexes. Moreover, males must identify the few sporadically entering females by detecting the females’ faint flight tones. We show that the Anopheles circadian clock not only ensures a tight synchrony of male and female activity but also helps sharpen the males’ acoustic detection system: By raising their flight tones to 1.5 times the female flight tone, males enhance the audibility of females, specifically at swarm time. Previously reported “harmonic convergence” events are only a random by-product of the mosquitoes’ flight tone variance and not a signature of acoustic interaction between males and females. The flight tones of individual mosquitoes occupy narrow, partly non-overlapping frequency ranges, suggesting that the audibility of individual females varies across males

Hitting the right note at the right time: circadian control of audibility in Anopheles mosquito mating swarms is mediated by flight tones

[Figure: see text]

Paratransgenesis to control malaria vectors: a semi-field pilot study

BACKGROUND: Malaria still remains a serious health burden in developing countries, causing more than 1 million deaths annually. Given the lack of an effective vaccine against its major etiological agent, Plasmodium falciparum, and the growing resistance of this parasite to the currently available drugs repertoire and of Anopheles mosquitoes to insecticides, the development of innovative control measures is an imperative to reduce malaria transmission. Paratransgenesis, the modification of symbiotic organisms to deliver anti-pathogen effector molecules, represents a novel strategy against Plasmodium development in mosquito vectors, showing the potential to reduce parasite development. However, the field application of laboratory-based evidence of paratransgenesis imposes the use of more realistic confined semi-field environments. METHODS: Large cages were used to evaluate the ability of bacteria of the genus Asaia expressing green fluorescent protein (Asaia (gfp)), to diffuse in Anopheles stephensi and Anopheles gambiae target mosquito populations. Asaia (gfp) was introduced in large cages through the release of paratransgenic males or by sugar feeding stations. Recombinant bacteria transmission was directly detected by fluorescent microscopy, and further assessed by molecular analysis. RESULTS: Here we show the first known trial in semi-field condition on paratransgenic anophelines. Modified bacteria were able to spread at high rate in different populations of An. stephensi and An. gambiae, dominant malaria vectors, exploring horizontal ways and successfully colonising mosquito midguts. Moreover, in An. gambiae, vertical and trans-stadial diffusion mechanisms were demonstrated. CONCLUSIONS: Our results demonstrate the considerable ability of modified Asaia to colonise different populations of malaria vectors, including pecies where its association is not primary, in large environments. The data support the potential to employ transgenic Asaia as a tool for malaria control, disclosing promising perspective for its field application with suitable effector molecules

Human CD8+T cells mediate protective immunity induced by a human malaria vaccine in human immune system mice

A number of studies have shown that CD8+ T cells mediate protective anti-malaria immunity in a mouse model. However, whether human CD8+ T cells play a role in protection against malaria remains unknown. We recently established human immune system (HIS) mice harboring functional human CD8+ T cells (HIS-CD8 mice) by transduction with HLA-A∗0201 and certain human cytokines using recombinant adeno-associated virus-based gene transfer technologies. These HIS-CD8 mice mount a potent, antigen-specific HLA-A∗0201-restricted human CD8+ T-cell response upon immunization with a recombinant adenovirus expressing a human malaria antigen, the Plasmodium falciparum circumsporozoite protein (PfCSP), termed AdPfCSP. In the present study, we challenged AdPfCSP-immunized HIS-CD8 mice with transgenic Plasmodium berghei sporozoites expressing full-length PfCSP and found that AdPfCSP-immunized (but not naïve) mice were protected against subsequent malaria challenge. The level of the HLA-A∗0201-restricted, PfCSP-specific human CD8+ T-cell response was closely correlated with the level of malaria protection. Furthermore, depletion of human CD8+ T cells from AdPfCSP-immunized HIS-CD8 mice almost completely abolished the anti-malaria immune response. Taken together, our data show that human CD8+ T cells mediate protective anti-malaria immunity in vivo

Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells

Purpose Human melanoma is a highly aggressive incurable cancer due to intrinsic cellular resistance to apoptosis, reprogramming, proliferation and survival during tumour progression. Sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, plays a role in carcinogenesis in many cancer types. However, the cytotoxic molecular mechanisms and gene expression profiles promoted by SFN in human melanoma remain unknown. Methods Three different cell lines were used: two human melanoma A375 and 501MEL and human epidermal melanocytes (HEMa). Cell viability and proliferation, cell cycle analysis, cell migration and invasion and protein expression and phosphorylation status of Akt and p53 upon SFN treatment were determined. RNA-seq of A375 was performed at different time points after SFN treatment. Results We demonstrated that SFN strongly decreased cell viability and proliferation, induced G2/M cell cycle arrest, promoted apoptosis through the activation of caspases 3, 8, 9 and hampered migration and invasion abilities in the melanoma cell lines. Remarkably, HEMa cells were not affected by SFN treatment. Transcriptomic analysis revealed regulation of genes involved in response to stress, apoptosis/cell death and metabolic processes. SFN upregulated the expression of pro-apoptotic genes, such as p53, BAX, PUMA, FAS and MDM2; promoted cell cycle inhibition and growth arrest by upregulating EGR1, GADD45B, ATF3 and CDKN1A; and simultaneously acted as a potent inhibitor of genotoxicity by launching the stress-inducible protein network (HMOX1, HSPA1A, HSPA6, SOD1). Conclusion Overall, the data show that SFN cytotoxicity in melanoma derives from complex and concurrent mechanisms during carcinogenesis, which makes it a promising cancer prevention agent

Topiramate-associated acute glaucoma in a migraine patient receiving concomitant citalopram therapy: a case-report

We describe the case of a 34 year-old man with diagnosis of migraine with and without aura that developed myopia and acute glaucoma after 7 days of treatment with topiramate. The patient had also been taking citalopram daily for two months. Both topiramate and citalopram have been related to the increase of intraocular pressure and the development of glaucoma. We can't exclude that in this patient citalopram caused an increase of the ocular pressure in dose-dependent manner, facilitating topiramate-induced glaucoma. We recommend to pay particular attention in prescribing of topiramate in migraine patients who are already under treatment with citalopram or other antidepressants with a similar mechanisms of action

Transition of plasmodium sporozoites into liver stage-like forms is regulated by the RNA binding protein pumilio

Many eukaryotic developmental and cell fate decisions that are effected post-transcriptionally involve RNA binding proteins as regulators of translation of key mRNAs. In malaria parasites (Plasmodium spp.), the development of round, non-motile and replicating exo-erythrocytic liver stage forms from slender, motile and cell-cycle arrested sporozoites is believed to depend on environmental changes experienced during the transmission of the parasite from the mosquito vector to the vertebrate host. Here we identify a Plasmodium member of the RNA binding protein family PUF as a key regulator of this transformation. In the absence of Pumilio-2 (Puf2) sporozoites initiate EEF development inside mosquito salivary glands independently of the normal transmission-associated environmental cues. Puf2- sporozoites exhibit genome-wide transcriptional changes that result in loss of gliding motility, cell traversal ability and reduction in infectivity, and, moreover, trigger metamorphosis typical of early Plasmodium intra-hepatic development. These data demonstrate that Puf2 is a key player in regulating sporozoite developmental control, and imply that transformation of salivary gland-resident sporozoites into liver stage-like parasites is regulated by a post-transcriptional mechanism