Stevens-Johnson syndrome (SJS) is a rare and severe immunologic phenomenon characterized by rash and mucous membrane disease. SJS may be triggered by medications and, less commonly, by infections such as Mycoplasma pneumoniae (Mp). Outbreaks of SJS are exceedingly rare.

We describe the largest SJS outbreak reported in children, which was also Mp-associated. In the first case-control study of this disease, we identify predictors of Mp-associated SJS versus non–Mp-associated SJS, including fewer skin lesions, pneumonia, and elevated erythrocyte sedimentation rate. (Read the full article)

Late preterm infants, compared with full-term infants, have less proficiency in reading and math at school age, with increased need for individualized educational plans and special education services. They also have lower cognitive performance on standardized IQ exams.

Late preterm infants have worse outcomes at school entry, and development is variable during the preschool years, so socioeconomic status, language spoken in the home, maternal education, maternal race, and being a late preterm infant have a large impact. (Read the full article)

High bilirubin levels are associated with sensorineural hearing loss. Exchange transfusions are recommended when bilirubin levels reach certain thresholds. However, the relative and excess risks of hearing loss in infants with bilirubin levels at/above exchange transfusion thresholds are unknown.

In this Northern California population of term and late preterm infants, elevated bilirubin levels were not associated with an increased risk of sensorineural hearing loss unless the levels were at least 10 mg/dL above exchange transfusion thresholds. (Read the full article)

Neurodevelopmental outcomes after cardiac surgery in early life provide critical information for understanding and improving care. Studies show these children are at risk for arterial ischemic stroke and acquired brain injury; further characterization of motor impairment is needed.

This study focuses on the presence of chronic neuromotor disabilities including cerebral palsy and motor impairments after acquired brain injury in children surviving early complex cardiac surgery, providing information on the frequency, characteristics, and predictors that may assist in prevention. (Read the full article)

Small for gestational age neonates (weight <10th percentile) are at risk for neutropenia during the first days after birth. However, the duration, responsible mechanism, and outcomes of this variety of neonatal neutropenia are not precisely known.

Six percent of small for gestational age neonates had neutrophils <1000/μL, with an average neutropenia duration of 7 days. Neutropenia was more closely linked with small for gestational age status than maternal hypertension. This neutropenia is associated with elevated nucleated red blood cell count and increased odds of necrotizing enterocolitis. (Read the full article)

Previous studies have shown racial differences in invasive pneumococcal disease (IPD) rates. Recent studies demonstrated a national decline in IPD rates after 13-valent pneumococcal conjugate vaccine (PCV13) introduction. The impact of PCV13 on racial and regional differences in IPD rates among Tennessee children is unknown.

After introduction of PCV13, pediatric IPD rates, including disease due to antibiotic-resistant strains, declined substantially. Racial and regional differences in IPD rates were no longer significant. Our study illustrates the impact of PCV13 and the importance of continued IPD surveillance. (Read the full article)

"We can do better," Germany coach Silvia Neid concluded after her side joined France and Sweden in making it to the quarter-finals of the Olympic tournament in unspectacular fashion.

A group of Bulgarian teenagers and leaders took part in this year’s TeenStreet Europe for the first time and experienced God in new ways.

Lyon extended their record tally of final wins to six while their run of four victories in a row is twice the next best.

03 November 2016 This briefing analyses the characteristics of EU nationals living in Scotland, based on the 2015 Annual Population Survey. The briefing also reviews evidence relating to the economic contribution of migrants.

United Kingdom and European Union negotiators were working through Sunday as the outcome of Brexit talks hung in the balance.

The heads of Deutsche Bank and Siemens, two of Germany’s biggest companies, are among a dwindling number of high-profile delegates still scheduled to attend an investment conference in Saudi Arabia following the disappearance of dissident journalist Jamal Khashoggi.

Finance ministers meeting in Brussels will try to push forward a legislative proposal for a levy on the European sales of companies with a global annual revenue of $853m or more, such as Facebook, Alphabet and Amazon.

Penn State and the Palmer Museum of Art mourn the loss of dear friend, generous donor, and loyal champion John P. Driscoll, who died from complications due to COVID-19 on Friday, April 10. Driscoll, owner of Driscoll Babcock Galleries in New York, was a longtime friend and supporter of the Palmer Museum and will be remembered for his role as a leader, gracious mentor and trusted adviser, as well as for the expansive gifts he made to the collection and to his alma mater, Penn State.

Allon Raiz is CEO of business incubator Raizcorp. In this series of articles, he offers entrepreneurs advice on surviving a crisis.

Now, more than ever, we need to be listening for market signals and ensuring that our businesses are primed to both take advantage of opportunities and to mitigate risks, says Allon Raiz.

La rechute est le risque majeur des pays post-conflit et l'une des principales raisons de cette rechute s'appelle la corruption.

Die Zentralafrikanische Republik kennzeichnet eine lange Geschichte der Instabilität. Wie die jüngste Krise gehandhabt wurde, deutet indessen auf eine neue sicherheitspolitische Konstellation hin.

Die Zentralafrikanische Republik trägt einen nüchternen, doch gerade in jüngster Zeit allzu treffenden Namen. Denn sie scheint tatsächlich zum Zentrum oder zumindest zum Ballungsraum all der vielfältigen Probleme geworden zu sein, unter denen der Kontinent seit langem leidet und von denen sich aber mittlerweile viele afrikanische Länder befreien können. Da sind die Konflikte um den Abbau wertvoller Ressourcen, besonders von Diamanten. Aus einer kleinen Elite ist eine Reihe unfähiger politischer Führer hervorgegangen. Es mehren sich Konflikte zwischen nomadischen Stämmen und der sesshaften Bevölkerung, was Ursachen auch im Klimawandel hat, und zu neuen Konkurrenzen zwischen den beiden Bevölkerungsteilen führt. Aus Rivalitäten zwischen ethnischen Gemeinschaften erwuchsen blutige Kämpfe und neue Feindschaften: die altbekannte, berüchtigte Kombination von historischen Altlasten und politischem Opportunismus schuf Konflikte auf der Grundlage religiöser Gruppenzugehörigkeit, wie jetzt zwischen Christen und Muslimen. Die schwierige Lage hat dem Land international eine erhöhte, aber nicht immer nützliche Aufmerksamkeit verschafft: des Nachbarlands Tschad, der Zentralafrikanischen Wirtschaftsgemeinschaft, von der Afrikanischen Union und von den Vereinten Nationen. Außerdem von multilateralen und Nicht­regierungs­organisationen, von der ehemaligen Kolonialmacht Frankreich und von weiteren internationalen Akteuren wie den Vereinigten Staaten, Südafrika, der Europäischen Union und unlängst auch von Deutschland. In kürzester Zeit ist die Zentralafrikanische Republik zu trauriger Berühmtheit gelangt. Doch so stark die Aufmerksamkeit auch gestiegen ist, so viel Wissen ist über dieses Land nachzuholen – Wissen, das man braucht, um vernünftig zu handeln. Mein Kollege Thibaud Lesueur und ich haben in den vergangenen drei Jahren viele Monate in der Zen­tralafrikanischen Republik verbracht. Wir konnten vor Ort beobachten, wie der Staat erst allmählich, dann rasant auseinanderfiel. Wir verfolgten, wie ein französisches Expeditionskorps, die Operation Sangaris, den afrikanischen Truppen zu Hilfe kam, um noch eine Spur von Ordnung vor dem drohenden Chaos zu retten, wie es nur eine gut ausgestattete Berufsarmee kann. Wir sahen, wie aus der einst stabilen Bevölkerung eine Generation plündernder Krieger hervorging. Und wir erlebten, wie aus Bangui, der Hauptstadt am Ubangi Fluss mit 750 000 Einwohnern, ein Schauplatz von Lynchjustiz wurde, was 90 Prozent der muslimischen Bevölkerung in die Flucht trieb. Ein Opfer dieser Selbstjustiz wurde auch Jean-Emmanuel Ndjaroua, ein Mitglied des nationalen Übergangsrats. Er machte im Februar den verhängnisvollen Fehler, öffentlich zu Toleranz und Frieden aufzurufen, und wurde auf offener Straße erschossen. Die große Herausforderung besteht nun darin zu verhindern, dass aus Tausenden viele Zehntausende Tote werden. Noch besteht Hoffnung, dass ein solches Blutvergießen vermieden werden kann. Die neue Regierung unter Präsidentin Catherine Samba-Panza hat Potenzial, und die von der Zentralafrikanischen Wirtschaftsgemeinschaft zügig entsandten – aber zu schwachen – Truppen hat man durch eine hoffentlich zielgerichtetere Mission unter Führung der ­Afrikanischen Union ersetzt. Unter der Federführung Frankreichs hat der UN-Sicherheitsrat am 10. April eine Resolution zum Einsatz einer neuen UN-Friedensmission beschlossen. Diese sieht vor, dass die Truppen der Afrikanischen Union im September 2014 unter das Kommando der Vereinten Nationen gestellt werden und die Zahl der internationalen Friedenssoldaten nahezu verdoppelt wird. Die Europäische Union hat derweil für Mai die Stationierung einer „Überbrückungsmission“ angekündigt. Diese Einsatzverpflichtungen sind mehr als bloße Versprechen, aber sie bleiben dennoch hinter dem zurück, was man als entschiedenes Handeln bezeichnen würde. Was also ist zu tun? Vertrauen zwischen den Religionen Es ist entscheidend, zwischen den Bevölkerungsteilen wieder Vertrauen aufzubauen. Der Imam, der Erzbischof und Vertreter der anderen christlichen Kirchen in Bangui arbeiten bereits eng zusammen, aber ihre Anstrengungen sind bisher auf die Hauptstadt begrenzt – aus der fast alle Muslime geflohen sind. Ein interkonfessioneller Dialog und eine Versöhnungskampagne müssen an der Basis beginnen und mithilfe der Übergangsregierung und ihrer internationalen Unterstützer auf die Provinzen ausgeweitet werden. Die Bausteine dafür existieren bereits – im Westen des Landes beispielsweise sind die verbliebenen muslimischen Flüchtlinge mehrheitlich bei christlichen Missionen untergekommen. Erinnern wir uns: Religiöse Gruppenzugehörigkeiten sind noch nicht lange eine Konfliktursache in der Zentralafrikanischen Republik. Zwei frühere Präsidenten, Bokassa und Patassé, konvertierten zum Islam, und diverse ethnische Gruppen setzen sich aus Christen und Muslimen zusammen. Bis heute sind im vorwiegend muslimischen nordöstlichen Distrikt, in dem sich auch viele aus Bangui vertriebene Krieger aufhalten, die Christen des Saraa-Stammes (zu dem auch viele Muslime gehören) nicht angegriffen worden, und auch nicht das zahlenmäßig große christliche Volk der Banda in Bria. In Bangui entstand als Reaktion auf die Morde eine Nichtregierungsorganisation, Les Frères Centrafricains, die über Aufkleber an Taxis zur Versöhnung aufrief. Junge Christen taten sich zusammen, um gemeinsam Moscheen vor Angriffen zu beschützen. Ankurbelung der Wirtschaft Die Wirtschaft des Landes muss neu belebt werden. Die wichtigsten Exportgüter des Landes sind Holz und Diamanten – und der Handel mit diesen Gütern setzt Sicherheit voraus. Von den fünf privaten Firmen, die bislang die Holzindustrie dominierten, arbeiten nur noch zwei. Ein Angestellter vor Ort erzählte uns, wie sein Betrieb zuerst Anfang 2013 von der Präsidentengarde durchsucht wurde, daraufhin von den muslimisch dominierten Séléka-Rebellen und schließlich von der prochristlichen Anti-Balaka-Bewegung, und wie alle von ihnen Fahrzeuge stahlen. Der Diamantenhandel ist ebenfalls in eine schwere Schieflage geraten, denn die Händler waren fast ausschließlich Muslime. Mit Beginn des Gegenaufstands der Anti-Balaka-Milizen flohen sie aus den Städten, ihre Geschäfte wurden geplündert. Auf lange Sicht muss der Staat seine Kontrolle über die Diamantenfelder wiederherstellen und für die Sicherheit der Händler sowie die Transparenz der Handelswege sorgen. Dazu müssen Zivilbeamte und Polizei eingesetzt werden. Zum jetzigen Zeitpunkt können allein Friedenstruppen dafür sorgen, dass der Handel wieder sicher aufgenommen werden kann. Der bedeutendste nicht exportorientierte Wirtschaftszweig des Landes ist die landwirtschaftliche Selbstversorgung. Auch sie leidet unter der problematischen Sicherheitslage, besonders dort, wo Nomaden und Farmer in Konkurrenz um Land aufeinandertreffen. Die Wanderungen der nomadischen Hirten aus dem Tschad im Norden in die Zentralafrikanische Republik müssen dringend unter eine von allen Seiten ausgehandelte Regelung gestellt werden, von der Art, wie sie in Niger und Tschad bereits erfolgreich ist. In den größeren Städten des Landes muss indessen Arbeit für die ­dortigen Kämpfer geschaffen werden. In der Hauptstadt Bangui herrscht Gewalt. Dort wird neben einer verbesserten Sicherheitslage dringend mehr Beschäftigung für die Jugendlichen gebraucht, damit diese eine Alternative zu den Milizen finden, die ihnen bisher „Arbeit“ verschafft haben. Die Hauptstadt und weitere Landesteile leiden unter infrastrukturellen Problemen, die durch beschäftigungsintensive Maßnahmen zu lösen wären, für die ungelernte und angelernte Arbeitskräfte eingesetzt werden können. Sicherheit Um die Sicherheit im Land wiederherzustellen, muss die Afrikanische Union mit den Vereinten Nationen zusammenarbeiten. Die neue UN-­Resolution will aus den 6000 AU-Soldaten UN-Blauhelme machen, aber Streit um Zuständigkeiten könnte die Umsetzung erschweren. Auch Frankreich und die Europäische Union müssen eine Grundlage zur Zusammenarbeit finden. Hier wird vermutlich Deutschland eine Schlüsselrolle spielen. Die deutsche Koalitionsregierung hat den Versuch gestartet, die deutsch-französische ­Zusammenarbeit neu zu beleben, gerade auf außenpolitischem Gebiet. Im April sprach Bundeskanzlerin Angela Merkel von Frankreich und Deutschland als „Motor“ der Beziehungen zwischen der EU und Afrika, und Frankreichs Staatspräsident François Hollande unterstrich die „besondere Freundschaft“ beider Länder. Deutschland hat sich in bisher nicht gekannter Weise verpflichtet, in Mali und der Zentralafrikanischen Republik militärische Hilfen bereitzustellen – dies soll jeweils in enger Abstimmung mit Frankreich geschehen. Hinzu kommt eine bedeutsame entwicklungspolitische Unterstützung. Diese französisch-deutsche Führung hat aus einem vagen Plan ein handfestes Unternehmen gemacht; mittlerweile haben sich Estland, Finnland, Frankreich, Deutschland, Italien, Lettland, Litauen, Luxemburg, Polen, Portugal, Schweden, Spanien, Großbritannien sowie Georgien zur Mission bekannt. Der Großteil der Truppen wird von Estland, Frankreich, Georgien, Polen und Spanien gestellt. Deutschland legt seinen Schwerpunkt auf den strategischen Lufttransport, Großbritannien kümmert sich um logistische Fragen und Italien um die Technik. Selbst wenn die EU-Überbrückungsmission Realität wird und sich die Beziehungen zur Afrikanischen Union verbessern, wird es für den ­Sicherheitsrat der Vereinten Nationen und die UN-Organisationen sehr schwierig werden, erfolgreich eine Blauhelmtruppe in der Zentralafrikanischen Republik zu etablieren. Die Vereinten Nationen müssen die Frage beantworten, welche ihrer Mitgliedstaaten die Truppen stellen. Dabei sollten sie Tschad außen vor lassen. Das Land ist schon zu sehr in die Angelegenheiten der Zentralafrikanischen Republik verstrickt und hat seine Friedensmission Anfang April aufgekündigt, nachdem tschadische Soldaten beschuldigt wurden, für den Tod von Zivilisten verantwortlich zu sein. Die Befehlsgewalt über die Truppen wird zwar formal im September von der AU auf die UN übergehen. Aber praktisch wird die UN-Mission wahrscheinlich nicht vor Ende des Jahres in vollem Umfang anlaufen. Dabei erfordert die Entwaffnung der Milizen schnelles Handeln: Die verbliebene muslimische Bevölkerung in Bangui hat sich in der PK5 genannten muslimischen Enklave bewaffnet, und auch die Anti-Balaka-Milizen haben bisher keine Probleme, in der Hauptstadt an Waffen zu kommen – obwohl Tausende französische und afrikanische Friedenssoldaten durch die Straßen der Hauptstadt patrouillieren. Der muslimische Bürgermeister von Banguis drittem Bezirk, zu dem auch PK5 gehört, sagte im März: „Wenn wir PK5 verlassen, um in ein benachbartes Gebiet zu gehen, werden wir noch am gleichen Tag getötet.“ Die Franzosen verpassten im Dezember und Januar die Gelegenheit zur weitgehenden Entwaffnung der Séléka-Truppen, als diese noch in vier Lagern festgehalten wurden. Eine UN-Mission wird es mit der Entwaffnung nicht leichter haben als die Franzosen. Der Plan der Vereinten Nationen sieht vor, dass zuerst Soldaten stationiert werden, dann eine funktionierende Polizei aufgebaut wird, und dann ein Justizsystem. Die größten Schwachstellen sind die Soldaten und Geld: Von beiden gibt es viel zu wenig. Ein strategischer Fahrplan Die Übergangsregierung der Zentralafrikanischen Republik wie auch die internationale Gemeinschaft brauchen dringend einen Plan. Auf nationaler Ebene hatte es Ende vergangenen Jahres einen gegeben – einen mangelhaften, aber immerhin etwas. Die neue Regierung unter Catherine Samba-Panza ist die dritte Regierung innerhalb eines Jahres, aber sie ist vielversprechend. Die meisten wichtigen Ministerien sind mit Technokraten statt mit Parteigenossen besetzt. Präsidentin Samba-Panza hat bereits zu Beginn ihrer Amtszeit die Bedeutung von Justiz und Versöhnung betont. In einem Gespräch im April mit Crisis-Group-Präsidentin Louise Arbour bestätigte sie dies und unterstrich, dass ihr Land auf die Hilfe ausländischer Richter angewiesen sei, um ein effektives Justizsystem aufzubauen. All dies gibt Anlass zur Hoffnung. Was man jedoch für die erweiterte internationale Intervention braucht, sind eine kohärente Führung, strategisches Denken sowie einen gemeinsamen Stabilisierungsplan, der es der Übergangsregierung ermöglicht, mittel- und langfristige Prioritäten zu setzen. Die internationale Kontaktgruppe sowie der jüngste Bericht des UN-Generalsekretärs betonen zu Recht, wie wichtig es ist, die Sicherheit im Land wiederherzustellen, die staatlichen Dienste zu reaktivieren und Wahlen vorzubereiten. Aber sie gehen nicht auf die tieferliegenden Probleme ein, insbesondere nicht auf den wirtschaftlichen Niedergang, der die Hauptursache für den Staatszerfall ist. Jemand – beispielsweise die EU oder Mitglieder der internationalen Kontaktgruppe – sollte dafür sorgen, dass die bisherigen Entwicklungs- und Aufbauprogramme überprüft werden. Eine solche Überprüfung ist aus zwei Gründen wichtig: erstens um zu verstehen, was bei der Reform des Sicherheitssektors, bei der Entwaffnung und Demobilisierung von Kämpfern sowie den Reintegrationsmaßnahmen schiefgelaufen ist. Und zweitens, um eine Stabilisierungsstrategie zu entwerfen, die die Wirtschaft in den Mittelpunkt stellt und das bietet, was zurzeit fehlt: neue Ideen für alte Probleme sowie eine langfristige Roadmap für die nationalen und internationalen Akteure. Auf diese Weise hätte man eine Grundlage für die angedachte Geberkonferenz im späteren Jahresverlauf und es wäre einfacher, eine sinnvolle Aufgabenverteilung unter den internationalen Akteuren zu verabreden. Ein solcher Rahmen ist unverzichtbar, um der neuen Regierung ein Fundament für ihre Herkulesaufgabe zu geben, nämlich aus dem Land wieder einen funktionierenden Staat zu machen und die Wirtschaft wieder aufzubauen, die Grundlage eines jeden zukunftsfähigen Staates. Bei dieser Überprüfung sollte man keine Zeit verlieren. Denn wie schon in der Vergangenheit könnte das ­Interesse der in­ternationalen Gemeinschaft an der Zentralafrikanischen Republik wieder schwinden. Gerade dies war in der Vergangenheit ein Hauptgrund dafür, dass sich die Probleme des Landes so häufig wiederholten. Geschieht das nicht, könnte es passieren, dass die geplante Überbrückungsmission schon in einem Jahr nur noch eine vage Erinnerung ist, dass der Fall Zentralafrikanische Republik Frankreich aufgebürdet wird, dass die afrikanischen Staaten gerade genug Mittel haben, um einzuschreiten, aber zu wenig, um irgendetwas langfristig zu stabilisieren, und dass die Übergangsregierung dann nur noch damit beschäftigt ist, die Fassade eines Staates aufrechtzuerhalten.

Tous les éléments d’une confrontation violente sont en place en Burundi. En observant les derniers développements, il semble que les éléments qui ont conduit par le passé à des massacres et à une longue guerre civile au début des années 1990 se remettent en place.

L’image de havre de paix dans une région en proie aux conflits dont bénéficiait le Cameroun a volé en éclats depuis l’irruption de Boko Haram en 2013 au nord du pays. Ce mouvement, devenu l’Etat islamique en Afrique de l’Ouest en mars 2015, revendique son affiliation à Daech. Néanmoins, l’apparition brutale et sanglante de ce djihadisme africain est moins liée à l’essor de Daech en Irak et en Syrie qu’aux bouleversements du paysage religieux de l’Afrique en général et du Cameroun en particulier.

Objectives. The aim of this study was to assess the safety of early oral switch (EOS) prior to 14 days for low-risk Staphylococcus aureus bacteremia (LR-SAB), which is the primary treatment strategy employed at our institution. Usually recommended therapy is 14 days of intravenous (IV) antibiotics.

Methods. All patients with SAB at our hospital were identified between 1 January 2014 and 31 December 2018. Those meeting low-risk criteria (healthcare-associated, no evidence of deep infection or demonstrated involvement of prosthetic material, and no further positive blood cultures after 72-hours) were included in the study. The primary outcome was occurrence of a SAB-related complication within 90 days.

Results. There were 469 SAB episodes during the study period, 100 (21%) of whom met inclusion criteria. EOS was performed in 84 patients. In this group, line infection was the source in 79%, methicillin-susceptible S. aureus caused 95% of SABs and 74% of patients received IV flucloxacillin. The median duration of IV and oral antibiotics in the EOS group was 5 (IQR 4-6) and 10 days (IQR 9-14), respectively. Seventy-one percent of patients received flucloxacillin as their EOS agent. Overall, 86% of oral step-down therapy was with beta-lactams. One patient (1%) undergoing EOS had SAB relapse within 90 days. No deaths attributable to SAB occurred within 90 days.

Conclusions. In this low MRSA prevalence LR-SAB cohort, EOS was associated with a low incidence of SAB-related complications. This was achieved with oral beta-lactam therapy in most patients. Larger prospective studies are needed to confirm these findings.

Leishmania major is the causative agent of cutaneous leishmaniasis (CL). No human vaccine is available for CL and current drug regimens present several drawbacks such as emerging resistance, severe toxicity, medium effectiveness, and/or high cost. Thus, the need for better treatment options against CL is a priority. In the present study, we validate the enzyme methionine aminopeptidase-1 (MetAP1), a metalloprotease that catalyzes the removal of N-terminal methionine from peptides and proteins, as a chemotherapeutic target against CL infection. The in vitro antileishmanial activity of eight novel MetAP1 inhibitors (OJT001-OJT008) were investigated. Three compounds OJT006, OJT007, and OJT008 demonstrated potent anti-proliferative effect in macrophages infected with L. major amastigotes and promastigotes at submicromolar concentrations, with no cytotoxicity against host cells. Importantly, the leishmanicidal effect was diminished by almost 10-fold in transgenic L. major promastigotes overexpressing MetAP1_LM in comparison to wild-type promastigotes. Furthermore, the in vivo activity of OJT006, OJT007, and OJT008 were investigated in L. major-infected BALB/c mice. In comparison to the control group, OJT008 significantly decreased footpad parasite load by 86%, and exhibited no toxicity against in treated mice. We propose MetAP1 inhibitor OJT008 as a potential chemotherapeutic candidate against CL infection caused by L. major infection.

The quinoline MK-571 is the most commonly used inhibitor of multidrug resistance protein-1 (MRP-1) but was originally developed as a cysteinyl leukotriene receptor 1 (CysLTR1) antagonist. While studying the modulatory effect of MRP-1 on anti-hepatitis C virus (HCV) direct acting-antivirals (DAA) efficiency, we observed an unexpected anti-HCV effect of compound MK-571 alone. This anti-HCV activity was characterized in Huh7.5 cells stably harboring a subgenomic genotype 1b replicon. A dose-dependent decrease of HCV RNA levels was observed upon MK-571 administration, with an EC₅₀ of 9±0.3 μM and a maximum HCV RNA level reduction of approximatively 1 Log₁₀. MK-571 also reduced the replication of the HCV full-length J6/JFH1 model in a dose-dependent manner. However, probenecid and apigenin homodimer (APN), two specific inhibitors of MRP-1, had no effect on HCV replication. In contrast, the CysLTR1 antagonists SR2640 increased HCV-SGR RNA levels in a dose-dependent manner, with a maximum increase of 10-fold. In addition, a combination of natural CysLTR1 agonist (LTD4) or antagonists (zafirlukast, cinalukast, and SR2640) with MK-571 completely reversed its antiviral effect, suggesting its anti-HCV activity is related to CysLTR1 rather to MRP-1 inhibition. In conclusion, we showed that MK-571 inhibits HCV replication in hepatoma cell cultures by acting as a CysLTR1 receptor antagonist, thus unraveling a new host-virus interaction in the HCV life cycle.

Objectives: Antibiotic combination therapy is used for severe infections caused by multidrug-resistant (MDR) Gram-negative bacteria. Yet, data of which combinations are most effective is lacking. This study aimed to evaluate the in vitro efficacy of polymyxin B in combination with 13 other antibiotics against four clinical strains of MDR Pseudomonas aeruginosa.

Methods: We evaluated the interactions of polymyxin B in combination with amikacin, aztreonam, cefepime, chloramphenicol, ciprofloxacin, fosfomycin, meropenem, minocycline, rifampicin, temocillin, thiamphenicol or trimethoprim by automated time-lapse microscopy using predefined cut-off values indicating inhibition of growth (≤10⁶ CFU/mL) at 24 h. Promising combinations were subsequently evaluated in static time-kill experiments.

Results: All strains were intermediate or resistant to polymyxin B, anti-pseudomonal β-lactams, ciprofloxacin and amikacin. Genes encoding β-lactamases (e.g., bla_PAO and bla_OXA-50) and mutations associated with permeability and efflux were detected in all strains. In the time-lapse microscopy experiments, positive interactions were found with 39 of 52 antibiotic combination/bacterial strain setups. Enhanced activity was found against all four strains with polymyxin B used in combination with aztreonam, cefepime, fosfomycin, minocycline, thiamphenicol and trimethoprim. Time kill experiments showed additive or synergistic activity with 27 of the 39 tested polymyxin B combinations, most frequently with aztreonam, cefepime, and meropenem.

Conclusion: Positive interactions were frequently found with the tested combinations, also against strains that harboured several resistance mechanisms to the single drugs and with antibiotics that are normally not active against P. aeruginosa. Further study is needed to explore the clinical utility of these combinations.

β-lactam resistance in Staphylococcus aureus limits treatment options. Stp1 and Stk1, a serine-threonine phosphatase and kinase respectively, mediate serine-threonine kinase (STK) signaling. Loss of function point mutations in stp1 were detected among laboratory passaged, β-lactam resistant S. aureus strains lacking mecA and blaZ, the major determinants of β-lactam resistance in the bacteria. Loss of Stp1 function facilitates β-lactam resistance of the bacteria.

Carbapenems are currently the preferred agents for the treatment of serious Acinetobacter infections. However, whether cefepime/cefpirome can be used to treat Acinetobacter bloodstream infection (BSI) if it is active against the causative pathogens is not clear. This study aimed to compare the efficacy of cefepime/cefpirome and carbapenem monotherapy in patients with Acinetobacter BSI. The population included 360 patients with monomicrobial Acinetobacter BSI receiving appropriate antimicrobial therapy admitted to four medical centres in Taiwan in 2012–2017. The predictors of 30-day mortality were determined by Cox regression analysis. The overall 30-day mortality rate in the appropriate antibiotic treatment group was 25.0% (90/360 patients), respectively. The crude 30-day mortality rates for cefepime/cefpirome and carbapenem therapy were 11.5% (7/61 patients) and 26.3% (21/80 patients), respectively. The patients receiving cefepime/cefpirome/carbapenem therapy were infected by Acinetobacter nosocomialis (51.8%), A. baumannii (18.4%) and A. pittii (12.1%). After adjusting for age, Sequential Organ Failure Assessment (SOFA) score, invasive procedures, and underlying diseases, cefepime/cefpirome therapy was not independently associated with a higher or lower 30-day mortality compared to the carbapenem therapy. SOFA score (hazard ratio [HR], 1.324; 95% confidence interval [CI], 1.137–1.543; P < 0.001) and neutropenia (HR, 7.060; 95% CI, 1.607–31.019; P = 0.010) were independent risk factors for 30-day mortality of patients receiving cefepime/cefpirome or carbapenem monotherapy. The incidence density of 30-day mortality for cefepime/cefpirome versus carbapenem therapy was 0.40% versus 1.04%. The therapeutic response of cefepime/cefpirome therapy was comparable to that of carbapenems among patients with Acinetobacter BSI receiving appropriate antimicrobial therapy.

Carbapenem-resistant Gram-negative pathogens are a critical public health threat and there is an urgent need for new treatments. Carbapenemases (β-lactamases able to inactivate carbapenems) have been identified in both serine β-lactamase (SBL) and metallo β-lactamase (MBL) families. The recent introduction of SBL carbapenemase-inhibitors has provided alternative therapeutic options. Unfortunately, there are no approved inhibitors of MBL-mediated carbapenem-resistance and treatment options for infections caused by MBL-producing Gram-negatives are limited. Here, we present ZN148, a zinc-chelating MBL-inhibitor capable of restoring the bactericidal effect of meropenem and in vitro clinical susceptibility to carbapenems in >98% of a large international collection of MBL-producing clinical Enterobacterales strains (n=234). Moreover, ZN148 was able to potentiate the effect of meropenem against NDM-1-producing Klebsiella pneumoniae in a murine neutropenic peritonitis model. ZN148 showed no inhibition of the human zinc-containing enzyme glyoxylase II at 500 μM and no acute toxicity was observed in an in vivo mouse model with cumulative dosages up to 128 mg/kg. Biochemical analysis showed a time-dependent inhibition of MBLs by ZN148 and removal of zinc ions from the active site. Addition of exogenous zinc after ZN148 exposure only restored MBL activity by ~30%, suggesting an irreversible mechanism of inhibition. Mass-spectrometry and molecular modelling indicated potential oxidation of the active site Cys221 residue. Overall, these results demonstrate the therapeutic potential of a ZN148-carbapenem combination against MBL-producing Gram-negative pathogens and that ZN148 is a highly promising MBL inhibitor, capable of operating in a functional space not presently filled by any clinically approved compound.

Ibrexafungerp (formerly SCY-078) is a semisynthetic triterpenoid and potent (1->3)-β-D-glucan synthase inhibitor. We investigated the in vitro activity, pharmacokinetics, and in vivo efficacy of ibrexafungerp (SCY) alone and in combination with anti-mould triazole isavuconazole (ISA) against invasive pulmonary aspergillosis (IPA). The combination of ibrexafungerp and isavuconazole in in vitro studies resulted in an additive and synergistic interactions against Aspergillus spp. Plasma concentration-time curves of ibrexafungerp were compatible with linear dose proportional profile. In vivo efficacy was studied in a well established persistently neutropenic NZW rabbit model of experimental IPA. Treatment groups included untreated rabbits (UC) and rabbits receiving ibrexafungerp at 2.5(SCY2.5) and 7.5(SCY7.5) mg/kg/day, isavuconazole at 40(ISA40) mg/kg/day, or combinations of SCY2.5+ISA40 and SCY7.5+ISA40. The combination of SCY+ISA produced in vitro synergistic interaction. There was significant in vivo reduction of residual fungal burden, lung weights, and pulmonary infarct scores in SCY2.5+ISA40, SCY7.5+ISA40, and ISA40-treatment groups vs that of SCY2.5-treated, SCY7.5-treated and UC (p<0.01). Rabbits treated with SCY2.5+ISA40 and SCY7.5+ISA40 had prolonged survival in comparison to that of SCY2.5-, SCY7.5-, ISA40-treated or UC (p<0.05). Serum GMI and (1->3)-β-D-glucan levels significantly declined in animals treated with the combination of SCY7.5+ISA40 in comparison to those treated with SCY7.5 or ISA40 (p<0.05). Ibrexafungerp and isavuconazole combination demonstrated prolonged survival, decreased pulmonary injury, reduced residual fungal burden, lower GMI and (1->3)-β-D-glucan levels in comparison to those of single therapy for treatment of IPA. These findings provide an experimental foundation for clinical evaluation of the combination of ibrexafungerp and an anti-mould triazole for treatment of IPA.

Background: Intensive care unit (ICU) patients may experience ceftriaxone underexposure but clinical outcomes data are lacking. The objective of this study was to determine the impact of ceftriaxone dosing on clinical outcomes amongst ICU patients without central nervous system (CNS) infection.

Methods: A retrospective study of ICU patients receiving intravenous, empiric ceftriaxone for non-CNS infections was conducted. Patients ≥18 years of age who received ≤2 grams of ceftriaxone daily for ≥72 hours were included and categorized as receiving ceftriaxone 1 gram or 2 grams daily. The primary, composite outcome was treatment failure: inpatient mortality and/or antibiotic escalation due to clinical worsening. Propensity score matching was performed based on the probability of receiving ceftriaxone 2 grams daily. Multivariable logistic regression determined the association between ceftriaxone dose and treatment failure in a propensity-matched cohort.

Results: A total of 212 patients were included in the propensity-matched cohort. The most common diagnoses (83.0%) were pneumonia and urinary tract infection. Treatment failure occurred in 17.0% and 5.7% of patients receiving 1 gram and 2 grams daily, respectively (p=0.0156). Overall inpatient mortality was 8.5%. Ceftriaxone 2 gram dosing was associated with a reduced likelihood of treatment failure (adjusted odds ratio=0.190; 95% confidence interval: 0.059 – 0.607). Other independent predictors of treatment failure included sequential organ failure assessment score (aOR 1.440, 95% CI 1.254 – 1.653) and creatinine clearance at 72 hours from ceftriaxone initiation (aOR 0.980, 95% CI (0.971 – 0.999).

Conclusions: Ceftriaxone 2 grams daily when used as appropriate antimicrobial coverage may be appropriate for ICU patients with lower mortality risk.

Mucormycosis is a life-threatening infection with high mortality that occurs predominantly in immunocompromised patients. Manogepix (MGX) is a novel antifungal that targets Gwt1, an early step in the conserved glycosylphosphotidyl inositol (GPI) post-translational modification pathway of surface proteins in eukaryotic cells. Inhibition of inositol acylation by MGX results in pleiotropic effects including inhibition of maturation of GPI-anchored proteins necessary for growth and virulence. MGX has been previously shown to have in vitro activity against some strains of Mucorales. Here we assessed the in vivo activity of the prodrug fosmanogepix, currently in clinical development for the treatment of invasive fungal infections, against two Rhizopus arrhizus strains with high (4.0 μg/ml) and low (0.25 μg/ml) minimum effective concentration (MEC) values. In both invasive pulmonary infection models, treatment of mice with 78 mg/kg or 104 mg/kg fosmanogepix, along with 1-aminobenzotriazole to enhance the serum half-live of MGX in mice, significantly increased median survival time and prolonged overall survival by day 21 post infection when compared to placebo. In addition, administration of fosmanogepix resulted in a 1-2 log reduction in both lung and kidney fungal burden. For the 104 mg/kg fosmanogepix dose, tissue clearance and survival were comparable to clinically relevant doses of isavuconazole (ISA), which is FDA approved for the treatment of mucormycosis. These results support continued development of fosmanogepix as a first in class treatment for invasive mucormycosis.

Telacebec (Q203) is a new anti-tubercular drug with extremely potent activity against Mycobacterium ulcerans. Here, we explored the treatment-shortening potential of Q203 alone or in combination with rifampin (RIF) in a mouse footpad infection model. The first study compared Q203 at 5 and 10 mg/kg doses alone and with rifampin. Q203 alone rendered most mouse footpads culture-negative in 2 weeks. Combining Q203 with rifampin resulted in relapse-free cure 24 weeks after completing 2 weeks of treatment, compared to a 25% relapse rate in mice receiving RIF+clarithromycin, the current standard of care, for 4 weeks.

The second study explored the dose-ranging activity of Q203 alone and with RIF, including the extended activity of Q203 after treatment discontinuation. The bactericidal activity of Q203 persisted for ≥ 4 weeks beyond the last dose. All mice receiving just 1 week of Q203 at 2-10 mg/kg were culture-negative 4 weeks after stopping treatment. Mice receiving 2 weeks of Q203 at 0.5, 2 and 10 mg/kg were culture-negative 4 weeks after treatment. RIF did not increase the efficacy of Q203. A pharmacokinetics sub-study revealed that Q203 doses of 2-10 mg/kg in mice produce plasma concentrations similar to those produced by 100-300 mg doses in humans, with no adverse effect of RIF on Q203 concentrations.

These results indicate the extraordinary potential of Q203 to reduce the duration of treatment necessary for cure to ≤ 1 week (or 5 doses of 2-10 mg/kg) in our mouse footpad infection model and warrant further evaluation of Q203 in clinical trials.

Objectives: Carbapenemase-producing Enterobacterales and specifically KPC-producing Klebsiella pneumoniae (KPC-Kp) are rapidly spreading worldwide. The prognosis of ventilator-associated pneumonia (VAP) caused by KPC-producing Klebsiella pneumoniae (KPC-Kp) is not well known. Our study tries to assess whether ventilator-associated pneumonia caused by a KPC-Kp strain is associated with higher all-cause mortality than if caused by carbapenem-susceptible isolates.

Study design and methods: This is a retrospective cohort study of patients with VAP due to K. pneumoniae from a 35-bed polyvalent Intensive Care Unit in a university hospital (> 40,000 annual admissions) between January 2012 and December 2016. Adjusted multivariate analysis was used to study the association of KPC-Kp with 30-day all-cause mortality (Cox regression).

Results. We analyze 69 cases of K. pneumoniae VAP of which 39 were produced by a KPC-Kp strain with high-level resistance to meropenem (MIC > 16 mg/mL). All-cause mortality at 30 days was 41% in the KPC-Kp group (16/39) and 33.3% in the carbapenem-susceptible cases (10/30). KPC-Kp etiology was not associated with higher mortality when controlled for confounders (adjusted hazard ratio [lsqb]HR[rsqb] 1.25; 95% CI: 0.46–3.41). Adequate targeted therapy (HR 0.03; 95% CI: <0.01–0.23) was associated with all-cause mortality.

Conclussion. Assuming the limitations due to the available sample size, the prognosis of VAP caused by KPC-Kp is similar to VAPs caused by carbapenem-susceptible K. pneumoniae when appropriate treatment is used.

Background. CLSI and EUCAST susceptibility breakpoints for voriconazole and C. albicans differ by one dilution (≤0.125 and ≤0.06 mg/l, respectively) whereas the epidemiological cutoff values (ECOFF/ECV) with both methodologies are the same (0.03 mg/L). We therefore determined the pharmacokinetic-pharmacodynamic (PK/PD) breakpoints of voriconazole against C. albicans for both methodologies with an in vitro PK/PD model, which was validated using existing animal PK/PD data.

Methods. Four clinical wild-type and non-wild-type C. albicans isolates (voriconazole MICs 0.008-0.125 mg/l) were tested in an in vitro PK/PD model. For validation purposes, mouse PK were simulated and in vitro PD were compared with in vivo outcome. Human PK were simulated and the exposure-effect relationship fAUC_0-24/MIC was described for EUCAST and CLSI24/48h methods. PK/PD breakpoints were determined using the fAUC_0-24/MIC associated with half-maximal activity (EI₅₀) and Monte Carlo simulation analysis.

Results. The in vitro 24h-PD EI₅₀ of voriconazole against C. albicans were 2.5-5 (1.5-17) fAUC/MIC. However, the 72h-PD were higher, 133 (51-347) fAUC/MIC for EUCAST and 94 (35-252) fAUC/MIC for CLSI. The mean (95% confidence interval) probability of target attainment (PTA) was 100(95-100)%, 97(72-100)%, 83(35-99)%, and 49(8-91)% and 100(97-100)%, 99(85-100)%, 91(52-100)% and 68(17-96)% for EUCAST and CLSI MICs 0.03, 0.06, 0.125, and 0.25 mg/L, respectively. Significantly, >95% PTAs were found for EUCAST/CLSI MICs ≤0.03 mg/ll. For MICs 0.06-0.125 mg/l trough levels 1-4 mg/ll would be required.

Conclusion. A PK/PD breakpoint of C. albicans voriconazole at the ECOFF/ECV of 0.03 mg/L was determined for both EUCAST/CLSI methods, indicating the need for breakpoint harmonization for the reference methodologies.

QPX7728 is an ultra-broad-spectrum boronic acid beta-lactamase inhibitor that demonstrates inhibition of key serine and metallo beta-lactamases at a nano molar range in biochemical assays with purified enzymes. The broad-spectrum inhibitory activity of QPX7728 observed in biochemical experiments translates into enhancement of the potency of many beta-lactams against strains of target pathogens producing beta-lactamases. The impact of bacterial efflux and permeability on inhibitory potency were determined using isogenic panels of KPC-3 producing isogenic strains of K. pneumoniae and P. aeruginosa and OXA-23-producing strains of A. baumannii with various combinations of efflux and porin mutations. QPX7728 was minimally affected by multi-drug resistance efflux pumps in either Enterobacteriaceae, or in non-fermenters such as P. aeruginosa or A. baumannii. In P. aeruginosa, the potency of QPX7728 was further enhanced when the outer membrane is permeabilized. The potency of QPX7728 in P. aeruginosa is not affected by inactivation of the carbapenem porin OprD. While changes in OmpK36 (but not OmpK35) reduced the potency of QPX7728 (8-16-fold), QPX7728 (4 μg/ml) nevertheless completely reversed KPC-mediated meropenem resistance in strains with porin mutations, consistent with a lesser effect of these mutations on the potency of QPX7728 compared to other agents. The ultra-broad-spectrum beta-lactamase inhibition profile combined with enhancement of the activity of multiple beta-lactam antibiotics with varying sensitivity to the intrinsic resistance mechanisms of efflux and permeability indicate QPX7728 is a useful inhibitor for use with multiple beta-lactam antibiotics.

Contezolid (MRX-I), a new oxazolidinone, is an antibiotic in development for treating complicated skin and soft tissue infections (cSSTI) caused by resistant Gram-positive bacteria. This was a thorough QT study conducted in 52 healthy subjects who were administered oral contezolid at a therapeutic (800 mg) dose, a supratherapeutic (1600 mg) dose, placebo, and oral moxifloxacin 400 mg in 4 separate treatment periods. The pharmacokinetic profile of contezolid was also evaluated. Time-point analysis indicated that the upper bounds of the two-sided 90% confidence interval (CI) for placebo-corrected change-from-baseline QTc (QTc) were <10 ms for the contezolid therapeutic dose at each time point. The upper bound of the 90% CI for QTc were slightly more than 10 ms with the contezolid supratherapeutic dose at 3 and 4 hours postdose, and the prolongation effect on the QT/QTc interval was less than that of the positive control, moxifloxacin 400 mg. At 3 and 4 h after the moxifloxacin dose, the moxifloxacin group met the assay sensitivity criteria outlined in ICH Guidance E14 with having a lower confidence bound ≥5 ms. The results of a linear exposure-response model which were similar to that of a time point analysis demonstrated a slightly positive relationship between contezolid plasma levels and QTcF interval with a slope of 0.227 ms per mg/L (90% CI: 0.188 to 0.266). In summary, contezolid did not prolong the QT interval at a therapeutic dose and may have a slight effect on QT interval prolongation at a supratherapeutic dose.

Polymyxins are increasingly used as the critical last-resort therapeutic options for multidrug-resistant gram-negative bacteria. Unfortunately, polymyxin resistance has increased gradually for the last few years. Although studies on mechanisms of polymyxin are expanding, system-wide analyses of the underlying mechanism for polymyxin resistance and stress response are still lacking. To understand how Klebsiella pneumoniae adapt to colistin (polymyxin E) pressure, we carried out proteomic analysis of Klebsiella pneumoniae strain cultured with different concentrations of colistin. Our results showed that the proteomic responses to colistin treatment in Klebsiella pneumoniae involving several pathways, including (i) gluconeogenesis and TCA cycle; (ii) arginine biosynthesis; (iii) porphyrin and chlorophyll metabolism; and (iv) enterobactin biosynthesis. Interestingly, decreased abundance of class A β-lactamases including TEM, SHV-11, SHV-4 were observed in cells treated with colistin. Moreover, we also present comprehensive proteome atlases of paired polymyxin-susceptible and -resistant Klebsiella pneumoniae strains. The polymyxin-resistant strain Ci, a mutant of Klebsiella pneumoniae ATCC BAA 2146, showed missense mutation in crrB. The crrB mutant Ci, which displayed lipid A modification with 4-amino-4-deoxy-L-arabinose (L-Ara4N) and palmitoylation, showed striking increases of CrrAB, PmrAB, PhoPQ, ArnBCADT and PagP. We hypothesize that crrB mutations induce elevated expression of the arnBCADTEF operon and pagP via PmrAB and PhoPQ. Moreover, multidrug efflux pump KexD, which was induced by crrB mutation, also contributed to colistin resistance. Overall, our results demonstrated proteomic responses to colistin treatment and the mechanism of CrrB-mediate colistin resistance, which may further offer valuable information to manage polymyxin resistance.

Third-generation cephalosporin (3GC)-resistant Enterobacteriaceae are classified as critical priority pathogens, with extended-spectrum β-lactamases (ESBLs) as principal resistance determinants. Enmetazobactam (formerly AAI101) is a novel ESBL inhibitor developed in combination with cefepime for empiric treatment of serious Gram-negative infections in settings where ESBLs are prevalent. Cefepime-enmetazobactam has been investigated in a phase 3 trial in patients with complicated urinary tract infections or acute pyelonephritis. This study examined pharmacokinetic-pharmacodynamic (PK-PD) relationships of enmetazobactam, in combination with cefepime, for ESBL-producing isolates of Klebsiella pneumoniae in 26-hour murine neutropenic thigh infection models. Enmetazobactam dose fractionation identified time above a free threshold concentration (fT > C_T) as the PK-PD index predictive of efficacy. Nine ESBL-producing isolates of K. pneumoniae, resistant to cefepime and piperacillin-tazobactam, were included in enmetazobactam dose-ranging studies. The isolates encoded CTX-M-type, SHV-12, DHA-1 and OXA-48 β-lactamases and covered a cefepime-enmetazobactam MIC range from 0.06 to 2 μg/ml. Enmetazobactam restored the efficacy of cefepime against all isolates tested. Sigmoid curve fitting across the combined set of isolates identified enmetazobactam PK-PD targets for stasis and for a 1-log₁₀ bioburden reduction of 8% and 44% fT > 2 μg/ml, respectively, with a concomitant cefepime PK-PD target of 40 – 60% fT > cefepime-enmetazobactam MIC. These findings support clinical dose selection and breakpoint setting for cefepime-enmetazobactam.

Mycobacterium abscessus lung infections remain difficult to treat. Recent studies have recognized the power of new combinations of antibiotics such as bedaquiline and imipenem although in vitro data have questioned this combination. We report that the efficacy of the bedaquiline plus imipenem treatment relies essentially on the activity of bedaquiline in a C3HeB/FeJ mice model of infection with a rough variant of M. abscessus. The addition of imipenem contributed at clearing the infection in the spleen.

Klebsiella pneumoniae that produce extended spectrum beta lactamases (ESBLs) are a persistent public health threat. There are relatively few therapeutic options and there is undue reliance on carbapenems. Alternative therapeutic options are urgently required. A combination of cefepime and the novel beta lactamase inhibitor enmetazobactam is being developed for treatment of serious infections caused by ESBL-producing organisms. The pharmacokinetics-pharmacodynamics (PK-PD) of cefepime-enmetazobactam against ESBL-producing K. pneumoniae was studied in a neutropenic murine pneumonia model. Dose ranging studies were performed. Dose fractionation studies were performed to define the relevant PD index for the inhibitor. The partitioning of cefepime and enmetazobactam into the lung was determined by comparing area under the concentration time curve (AUC) in plasma and epithelial lining fluid. The magnitude of drug exposure for cefepime-enmetazobactam required for logarithmic killing in the lung was defined using 3 ESBL-producing strains. Cefepime 100 mg/kg q8h i.v. had minimal antimicrobial effect. When this background regimen of cefepime was combined with enmetazobactam half-maximal effect was induced with enmetazobactam 4.71 mg/kg q8h i.v. The dose fractionation study suggest both fT>threshold and fAUC:MIC are potentially relevant PD indices. The AUC_ELF:AUC_plasma for cefepime and enmetazobactam was 73.4% and 61.5%, respectively. A ≥2-log kill in the lung was achieved with a plasma and ELF cefepime fT>MIC of ≥20% and enmetazobactam fT>2 mg/L of ≥20% of the dosing interval. These data and analyses provide the underpinning evidence for the combined use of cefepime and enmetazobactam for nosocomial pneumonia.

The human diseases caused by the fungal pathogens Cryptococcus neoformans and C. gattii are associated with high indices of mortality, and toxic and/or cost-prohibitive therapeutic protocols. The need for affordable antifungals to combat cryptococcal disease is unquestionable. Previous studies suggested benzimidazoles as promising anti-cryptococcal agents combining low cost and high antifungal efficacy, but their therapeutic potential has not been demonstrated so far. In this study, we investigated the antifungal potential of fenbendazole, the most effective anti-cryptococcal benzimidazole. Fenbendazole was inhibitory against 17 different isolates of C. neoformans and C. gattii at a low concentration. The mechanism of anti-cryptococcal activity of fenbendazole involved microtubule disorganization, as previously described for human parasites. In combination with fenbendazole, the concentrations of the standard antifungal amphotericin B required to control cryptococcal growth were lower than those required when this antifungal was used alone. Fenbendazole was not toxic to mammalian cells. During macrophage infection, the anti-cryptococcal effects of fenbendazole included inhibition of intracellular proliferation rates and reduced phagocytic escape through vomocytosis. Fenbendazole deeply affected the cryptococcal capsule. In a mice model of cryptococcosis, the efficacy of fenbendazole to control animal mortality was similar to that observed for amphotericin B. These results indicate that fenbendazole is a promising candidate for the future development of an efficient and affordable therapeutic tool to combat cryptococcosis.

Carbapenem pharmacokinetic profiles are significantly changed in critically ill patients because of the drastic variability of the patients' physiological parameters. Published population PK studies have mainly focused on specific diseases and the majority of these studies had small sample sizes. The aim of this study was to develop a population PK model of imipenem in critically ill patients that estimated the influence of various clinical and biological covariates and the use of Extracorporeal Membrane Oxygenation (ECMO) and Continuous Renal Replacement Therapy (CRRT). A two-compartment population PK model with Creatinine clearance (CrCL), body weight (WT), and ECMO as fixed effects was developed using the non-linear mixed effect model (NONMEM). A Monte Carlo simulation was performed to evaluate various dosing schemes and different levels of covariates based on the pharmacokinetic/pharmacodynamic index (f%T>MIC) for the range of clinically relevant minimum inhibitory concentrations(MICs). The results showed that there may be insufficient drug use in the clinical routine drug dose regimen, and 750mg Q6h could achieve a higher treatment success rate. The blood concentrations of imipenem in ECMO patients were lower than that of non-ECMO patients, therefore dosage may need to be increased. The dosage may need adjustment for patients with CrCL ≤ 70ml/min, but dose should be lowered carefully to avoid the insufficient drug exposure. Dose adjustment is not necessary for patients within the WT ranging from 50-80 kg. Due to the large variation in PK profile of imipenem in critically ill patients, TDM should be carried out to optimize drug regimens.

Clostridioides difficile, the leading cause of nosocomial infections, is an urgent health threat worldwide. The increased incidence and severity of disease, the high recurrence rates, and the dearth of effective anticlostridial drugs have created an urgent need for new therapeutic agents. In an effort to discover new drugs for treatment of Clostridioides difficile infections (CDIs), we investigated a panel of FDA-approved antiparasitic drugs against C. difficile and identified diiodohydroxyquinoline (DIHQ), an FDA-approved oral antiamoebic drug. DIHQ exhibited potent activity against 39 C. difficile isolates, inhibiting growth of 50% and 90% of these isolates at the concentrations of 0.5 μg/mL and 2 μg/mL, respectively. In a time-kill assay, DIHQ was superior to vancomycin and metronidazole, reducing a high bacterial inoculum by 3-log₁₀ within six hours. Furthermore, DIHQ reacted synergistically with vancomycin and metronidazole against C. difficile in vitro. Moreover, at subinhibitory concentrations, DIHQ was superior to vancomycin and metronidazole in inhibiting two key virulence factors of C. difficile, toxin production and spore formation. Additionally, DIHQ did not inhibit growth of key species that compose the host intestinal microbiota, such as Bacteroides, Bifidobacterium and Lactobacillus spp. Collectively, our results indicate that DIHQ is a promising anticlostridial drug that warrants further investigation as a new therapeutic for CDIs.

Probiotics might provide an alternative approach for the control of oral candidiasis. However, studies on the antifungal activity of probiotics in the oral cavity are based on the consumption of yogurt or other dietary products, and there is a necessary to use appropriate biomaterials and specific strains to obtain probiotic formulations targeting local oral administration. In this study, we impregnated gellan gum, a natural biopolymer used as a food-additive, with a probiotic and investigated its antifungal activity against Candida albicans. Lactobacillus paracasei 28.4, a strain recently isolated from the oral cavity of a caries-free individual, was incorporated in several concentrations of gellan gum (0.6% to 1%). All tested concentrations could incorporate L. paracasei cells while maintaining bacterial viability. Probiotic/gellan formulations were stable for 7 days when stored at room temperature or 4°C. Long-term storage of bacteria-impregnated gellan gum was achieved when L. paracasei 28.4 was lyophilized. The probiotic/gellan formulations provided a release of L. paracasei cells over 24 hours that was sufficient to inhibit the growth of C. albicans with effects dependent on the cell concentrations incorporated into gellan gum. The probiotic/gellan formulations also had inhibitory activity against Candida spp. biofilms by reducing the number of Candida spp. cells (p < 0.0001), decreasing the total biomass (p = 0.0003), and impairing hyphae formation (p = 0.0002), compared to the control group which received no treatment. Interestingly, probiotic formulation of 1% w/v gellan gum provided an oral colonization of L. paracasei in mice with approximately 6 log of CFU/mL after 10 days. This formulation inhibited the C. albicans growth (p < 0.0001), prevented the development of candidiasis lesions (p = 0.0013), and suppressed inflammation (p = 0.0006) when compared to the mice not treated in the microscopic analysis of the tongue dorsum. These results indicate that gellan gum is a promising biomaterial and can be used as a carrier system to promote oral colonization for probiotics that prevent oral candidiasis.

Brain infections with Cryptococcus neoformans are associated with significant morbidity and mortality. Cryptococcosis typically presents as meningoencephalitis or fungal mass lesions called cryptococcomas. Despite frequent in vitro discoveries of promising novel antifungals, the clinical need for drugs that can more efficiently treat these brain infections remains. A crucial step in drug development is the evaluation of in vivo drug efficacy in animal models. This mainly relies on survival studies or post-mortem analyses in large groups of animals, but these techniques only provide information on specific organs of interest at predefined time points. In this proof-of-concept study, we validated the use of non-invasive preclinical imaging to obtain longitudinal information on the therapeutic efficacy of amphotericin B or fluconazole monotherapy in meningoencephalitis and cryptococcoma mouse models. Bioluminescence imaging (BLI) enabled the rapid in vitro and in vivo evaluation of drug efficacy while complementary high-resolution anatomical information obtained by magnetic resonance imaging (MRI) of the brain allowed a precise assessment of the extent of infection and lesion growth rates. We demonstrated a good correlation between both imaging readouts and the fungal burden in various organs. Moreover, we identified potential pitfalls associated with the interpretation of therapeutic efficacy based solely on post-mortem studies, demonstrating the added value of this non-invasive dual imaging approach compared to standard mortality curves or fungal load endpoints. This novel preclinical imaging platform provides insights in the dynamic aspects of the therapeutic response and facilitates a more efficient and accurate translation of promising antifungal compounds from bench to bedside.

Gepotidacin, a triazaacenaphthylene bacterial type II topoisomerase inhibitor, is in development for treatment of uncomplicated urinary tract infection (uUTI). This Phase 2a study in female participants with uUTI evaluated the pharmacokinetics (primary objective), safety, and exploratory efficacy of gepotidacin. Eligible participants (N = 22) were confined to the clinic at baseline, received oral gepotidacin 1,500 mg twice daily for 5 days (on-therapy; Days 1 to 5), and returned to the clinic for test-of-cure (Days 10 to 13) and follow-up (Day 28±3). Pharmacokinetic, safety, clinical, and microbiological assessments were performed. Maximum plasma concentrations were observed approximately 1.5 to 2 hours postdose. Steady state was attained by Day 3. Urinary exposure over the dosing interval increased from 3,742 μg.h/ml (Day 1) to 5,973 μg.h/ml (Day 4), with trough concentrations of 322 to 352 μg/ml from Day 3 onward. Gepotidacin had an acceptable safety-risk profile with no treatment-limiting adverse events and no clinically relevant safety trends. Clinical success was achieved in 19 (86%) and 18 (82%) of 22 participants at test-of-cure and follow-up, respectively. Eight participants had a qualifying baseline uropathogen (growth; ≥10⁵ CFU/ml). A therapeutic (combined clinical and microbiological [no growth; <10³ CFU/ml]) successful response was achieved in 6 (75%) and 5 (63%) of 8 participants at test-of-cure and follow-up, respectively. Plasma area under the free-drug concentration-time curve over 24 hours at steady state divided by the MIC (fAUC_0-24/MIC) and urine AUC_0-24/MIC ranged from 6.99 to 90.5 and 1,292 to 121,698, respectively. Further evaluation of gepotidacin in uUTI is warranted. (NCT03568942)

Treatment of dogs naturally infected with Leishmania infantum using meglumine antimoniate (MA) encapsulated in conventional liposomes (LC) in association with allopurinol has been previously reported to promote marked reduction in the parasite burden in the main infection sites. Here, a new assay in naturally infected dogs was performed using a novel liposome formulation of MA consisting of a mixture of conventional and long-circulating (PEGylated) liposomes (LCP), with expected broader distribution among affected tissues of the mononuclear phagocyte system. Experimental groups of naturally infected dogs were as follows: LCP+Allop, receiving LCP intravenously as 2 cycles of 6 doses (6.5 mg Sb/kg/dose) at 4-day intervals, plus allopurinol at 30 mg/kg/12 h p.o. during 130 days; LC+Allop, receiving LC intravenously as 2 cycles of 6 doses (6.5 mg Sb/kg/dose), plus allopurinol during 130 days; Allop, treated with allopurinol only; non-treated control. Parasite loads were evaluated by quantitative PCR in liver, spleen and bone marrow and by immunohistochemistry in the ear skin, before, just after treatment and 4 months later. LCP+Allop and LC+Allop groups, but not the Allop group, showed significant suppression of the parasites in the liver, spleen and bone marrow 4 months after treatment, compared to the pre-treatment period or the control group. Only LCP+Allop group showed significantly lower parasite burden in the skin, in comparison to the control group. On the basis of clinical staging and parasitological evaluations, LCP formulation exhibited a more favorable therapeutic profile, when compared to LC one, being therefore promising for treatment of canine visceral leishmaniasis.

Pharmacokinetic changes are often seen in patients with severe infections. Administration by continuous infusion has been suggested to optimize antibiotic exposure and pharmacokinetic/pharmacodynamic (PK/PD) target attainment for β-lactams. In an observational study, unbound piperacillin concentrations (n=196) were assessed in 78 critically ill patients following continuous infusion of piperacillin/tazobactam (ratio 8:1). The initial dose of 8, 12 or 16 g (piperacillin component) was determined by individual creatinine clearance (CRCL). Piperacillin concentrations were compared to the EUCAST clinical breakpoint MIC for Pseudomonas aeruginosa (16 mg/L), and the following PK/PD targets were evaluated: 100% fT>1xMIC and 100% fT>4xMIC. A population pharmacokinetic model was developed using NONMEM 7.4.3 consisting of a one-compartment disposition model with linear elimination separated into non-renal and renal (linearly increasing with patient CRCL) clearances. Target attainment was predicted and visualized for all individuals based on the utilized CRCL dosing algorithm. The target of 100% fT>1xMIC was achieved for all patients based on the administered dose, but few patients achieved the target of 100% fT>4xMIC. Probability of target attainment for a simulated cohort of patients showed, that increasing the daily dose by 4 g increments (piperacillin component) did not result in substantially improved target attainment for the 100% fT>4xMIC target. To conclude, in patients with high CRCL combined with high-MIC bacterial infections, even a CI regimen with a daily dose of 24 g may be insufficient to achieve therapeutic concentrations.

Attention has been paid to H5N6 highly pathogenic avian influenza virus (HPAIV) because of its heavy burden on the poultry industry and human mortality. Since an influenza A virus carrying N6 neuraminidase (NA) has never spread in humans, the potential for H5N6 HPAIV to cause disease in humans and the efficacy of antiviral drugs against the virus need to be urgently assessed. We used non-human primates to elucidate the pathogenesis of H5N6 HPAIV as well as to determine the efficacy of antiviral drugs against the virus. H5N6 HPAIV infection led to high fever in cynomolgus macaques. The lung injury caused by the virus was severe with diffuse alveolar damage and neutrophil infiltration. In addition, an increase in IFN-α showed an inverse correlation with virus titers during the infection process. Oseltamivir was effective for reducing H5N6 HPAIV propagation, and continuous treatment with peramivir reduced virus propagation and severity of symptoms in the early stage. This study also showed the pathologically severe lung injury states in the cynomolgus macaques infected with H5N6 HPAIV, even in those that received early antiviral drug treatments, indicating the need for close monitoring and the need for further studies on the virus pathogenicity and new antiviral therapies.