Aim: Currently, we have limited armamentarium of antifungal agents against Mucorales. There is an urgent need to discover novel antifungal agents that are effective, safe and affordable. Materials & methods: In this study, the anti-Mucorale action of native lactoferrin (LF) and its functional fragments CLF, RR6 and LFcin against three common Mucorale species are reported. The synergistic action of LF with antifungal agents like amphotericin B, isavuconazole and posaconazole was analyzed using checkerboard technique. Results: All the three mucor species showed inhibition when treated with fragments. The checkerboard assay confirmed that native LF showed the best synergistic action against Mucorales in combination with Amphotericin B. Conclusion: These results highlight the potential therapeutic value of native LF against Mucorales.
Black fungus, or ‘mucormycosis’, is a dangerous fungal infection. Normally, it affects people with a weakened immune system. It is only treatable when diagnosed early. It spreads by breathing the fungus in, eating contaminated food or direct contact with an infected wound. There are not many medicines that can treat this type of fungus, so it is important to find new ones. In this study, we tested a natural protein called lactoferrin and some of its building blocks, called peptides, to see if they could stop the fungus from growing. Lactoferrin and its peptides could stop the fungus from growing, especially when used with a medicine called amphotericin B. This means that lactoferrin could potentially be a helpful treatment for this fungal infection.

A laboratory-based lateral flow (LF) test that utilizes up-converting reporter particles (UCP) for ultrasensitive quantification of Schistosoma circulating anodic antigen (CAA) in urine is a well-accepted test to identify active infection. However, this UCP-LF CAA test requires sample pre-treatment steps not compatible with field applications. Flow, a new low-cost disposable, allows integration of large-volume pre-concentration of urine analytes and LF detection into a single field-deployable device. We assessed a prototype Flow-Schistosoma (Flow-S) device with an integrated UCP-LF CAA test strip, omitting all laboratory-based steps, to enable diagnosis of active Schistosoma infection in the field using urine. Flow-S is designed for large-volume (5-20 mL) urine, applying passive paper-based filtration and antibody-based CAA concentration. Samples tested for schistosome infection were collected from women of reproductive age living in a Tanzania region where S. haematobium infection is endemic. Fifteen negative and fifteen positive urine samples, selected based on CAA levels quantified in paired serum, were analyzed with the prototype Flow-S. The current Flow-S prototype, with an analytical lower detection limit of 1 pg CAA/mL, produced results correlated with the laboratory-based UCP-LF CAA test. Urine precipitates occurred in frozen banked samples and affected accurate quantification; however, this should not occur in fresh urine. Based on the findings of this study, Flow-S appears suitable to replace the urine pre-treatment required for the laboratory-based UCP-LF CAA test, thus allowing true field-based applications with fresh urine samples. The urine precipitates observed with frozen samples, though less important given the goal of testing fresh urines, warrant additional investigation to evaluate methods for mitigation. Flow-S devices permit testing of pooled urine samples with applications for population stratified testing. A field test with fresh urine samples, a further optimized Flow-S device, and larger statistical power has been scheduled.

BACKGROUND: Fuzheng Huayu formula (FZHY), composed of Salvia miltiorrhiza Bunge, Cordyceps sinensis, the seed of Prunus persica (L.) Batsch, the pollen of Pinus massoniana Lamb, Gynostemma pentaphyllum (Thunb.) Makino and the fruit of Schisandra chinensis (Turcz.) Baill, is a Chinese herbal compound with demonstrated clinical benefits in liver fibrosis (LF). However, its potential mechanism and molecular targets remain to be elucidated.
OBJECTIVE: This study was designed to evaluate the anti-fibrotic role of FZHY in hepatic fibrosis and to elucidate the potential mechanisms.
METHODS: Network pharmacology was assayed to identify the interrelationships among compounds of FZHY, potential targets and putative pathways on anti-LF. Then the core pharmaceutical target for FZHY against LF was verified by serum proteomic analysis. Further in vivo and in vitro assays were performed to verify the prediction of the pharmaceutical network.
RESULTS: The network pharmacology analysis revealed that a total of 175 FZHY-LF crossover proteins were filtered into a protein-protein interaction (PPI) network complex and designated as the potential targets of FZHY against LF, and the Epidermal Growth Factor Receptor (EGFR) signaling pathway was further explored according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Then analytical studies were validated by carbon tetrachloride (CCl4)-induced model in vivo. We found FZHY could attenuate CCl4-induced LF, especially decrease p-EGFR expression in α-Smooth Muscle Actin (α-SMA)-positive hepatic stellate cell (HSC) and inhibit the downstream of the EGFR signaling pathway, especially Extracellular Regulated Protein Kinases (ERK) signaling pathway in liver tissue. We further demonstrate that FZHY could inhibit Epidermal Growth Factor (EGF)-induced HSC activation, as well as the expression of p-EGFR and the key protein of the ERK signaling pathway.
CONCLUSIONS: FZHY has a good effect against CCl4-induced LF. The action mechanism was associated with the down-regulation of the EGFR signaling pathway in activated HSCs.

UNASSIGNED: Polycystic ovary syndrome (PCOS) patients shows common features like increased insulin resistance and adiposity, which have been known to correlate with sympathetic hyperactivity.
UNASSIGNED: The aim of this study is to analyze the characteristics of heart rate variability in women with PCOS. To compare frequency domain parameters of heart rate variability (HRV) between women with PCOS and apparently healthy women. To study the impact of cardiometabolic parameters such as BMI and blood pressure on frequency-domain HRV parameters.
UNASSIGNED: A total of 30 women with PCOS aged 20 to 40 years (as per Rotterdam criteria) were enrolled as cases and 30 age-matched women having normal ovulatory cycles were enrolled as controls. HRV was recorded using an electrocardiography machine (ECG) machine. The following frequency-domain parameters were assessed: Total power, Very low frequency (VLF), VLF%, Low Frequency (LF), LF%, LF nu, High frequency (HF), HF%, HF nu, LF/HF ratio, short-term variability (SD1), and long-term variability (SD2), respectively.
UNASSIGNED: Mean age of cases was 28.03 ± 5.33 years. Mean BMI of PCOS women was 25.39 ± 2.69 kg/m2. A total of 18 (60%) had BMI >25 kg/m2. Cases had significantly higher BMI, waist hip ratio, and blood pressure as compared with controls. None of the controls had BMI >25 kg/m2. Majority of cases (66.7%) had systolic blood pressure/diastolic blood pressure (SBP/DBP) >130/85 mmHg as compared with only 6 (20%) of controls (P < 0.001). For different frequency domain parameters, no statistically significant difference between two groups was observed for VLF and LF. Mean VLF%, LF%, LF (nu), and LF/HF were significantly higher in cases as compared with controls. For all, the other mean value was significantly lower in cases as compared with that of controls (P < 0.05).
UNASSIGNED: Autonomic nervous system is affected by PCOS status of women, and sympathetic hyperactivity is seen.

The elimination of lymphatic filariasis (LF) is achieved through repeated mass drug administration (MDA) of anti-filarial medications, which interrupts transmission and prevents new infections. Accurate transmission assessments are critical to deciding when to stop MDA. Current methods for evaluating transmission may be insufficiently sensitive, resulting in post-MDA resurgence. We, therefore, evaluated potential diagnostic testing scenarios for post-MDA surveillance. Data were used from two surveys (a household cluster and a cohort) conducted in an area of Mandalay Region, Myanmar, with ongoing transmission following several rounds of MDA. First, age- and sex-adjusted seroprevalence were estimated for the area using the household survey. Next, three Bayesian networks were built from the combined datasets to compare antigens by immunochromatic testing (ICT) and/or Og4C3 enzyme-linked immunosorbent assay (ELISA) and antibody (Ab) detection methods (Wb123 or Bm14 Ab ELISA). The networks were checked for validity and then used to compare diagnostic testing scenarios. The adjusted prevalence from the household survey for antigen, Wb123 Ab and Bm14 Ab were 4.4% (95% CI 2.6-7.3%), 8.7% (5.96-12.5%) and 20.8% (16.0-26.6%), respectively. For the three networks, the True Skill Statistic and Area Under the Receiver Operating Characteristic Curve for antigen, Wb123 and Bm14 Ab were 0.79, 0.68 and 0.55; and 0.97, 0.92 and 0.80, respectively. In the Bayesian network analysis, a positive case was defined as testing positive to one or more infection markers. A missed result was therefore the probability of a positive case having a negative test result to an alternate marker. The probability of a positive case prior to any testing scenario was 17.4%, 16.8% and 26.6% for antigen, Wb123 Ab and Bm14 Ab, respectively. In the antigen-only testing scenario, the probability of a missed positive LF result was 5.2% for Wb123 and 15.6% for Bm14 Ab. The combination of antigen plus Bm14 Ab testing reduced the probability of missing a positive LF case as measured by Wb123 Ab to 0.88%. The combination of antigen plus Wb123 Ab was less successful and yielded an 11.5% probability of a missed positive result by Bm14 Ab testing. Across scenarios, there was a greater discordance between Bm14 and both antigen and Wb123 Ab in the 1-10 age group compared to older ages. These findings suggest that the addition of Bm14 Ab improves the sensitivity of LF testing for current or past infection. The combination of antigen plus Bm14 Ab should therefore be considered for inclusion in post-MDA surveillance to improve the sensitivity of transmission surveys and prevent the premature cessation of MDA.

The biologically active vitamin B12 derivates, methylcobalamin (MeCbl) and adenosylcobalamin (AdoCbl), are ubiquitous organometallic cofactors. In addition to their key roles in enzymatic catalysis, B12 cofactors have complex photolytic properties which have been the target of experimental and theoretical studies. With the recent discovery of B12-dependent photoreceptors, there is an increased need to elucidate the underlying photochemical mechanisms of these systems. This book chapter summarizes the photolytic properties of MeCbl- and AdoCbl-dependent enzymes with particular emphasis on the effect of the environment of the cofactor on the excited state processes. These systems include isolated MeCbl and AdoCbl as well as the enzymes, ethanolamine ammonia-lyase (EAL), glutamate mutase (GLM), methionine synthase (MetH), and photoreceptor CarH. Central to determining the photodissociation mechanism of each system is the analysis of the lowest singlet excited state (S1) potential energy surface (PES). Time-dependent density functional theory (TD-DFT), employing BP86/TZVPP, is widely used to construct such PESs. Regardless of the environment, the topology of the S1 PES of AdoCbl or MeCbl is marked by characteristic features, namely the metal-to-ligand charge transfer (MLCT) and ligand field (LF) regions. Conversely, the relative energetics of these electronic states are affected by the environment. Applications and outlooks for Cbl photochemistry are also discussed.

Lactoferrin (LF) is a multifunctional milk glycoprotein that promotes bone regeneration. Local delivery of LF at the bone defect site is a promising approach for enhancement of bone regeneration, but efficient systems for sustained local delivery are still largely missing. The aim of this study was to investigate the potential of the poloxamers for sustained delivery of LF to enhance local bone regeneration. The developed LF/poloxamer formulations were liquid at room temperature (20 °C) transforming to a sustained releasing gel depot at body temperature (37 °C). In vitro release studies demonstrated an initial burst release (~50%), followed by slower release of LF for up to 72 h. Poloxamer, with and without LF, increased osteoblast viability at 72 h (p < 0.05) compared to control, and the immune response from THP-1 cells was mild when compared to the suture material. In rat calvarial defects, the LF/poloxamer group had lower bone volume than the controls (p = 0.0435). No difference was observed in tissue mineral density and lower bone defect coverage scores (p = 0.0267) at 12 weeks after surgery. In conclusion, LF/poloxamer formulations support cell viability and do not induce an unfavourable immune response; however, LF delivery via the current formulation of LF200/poloxamer gel did not demonstrate enhanced bone regeneration and was not compatible with the rat calvarial defect model.

The Lymphatic Filariasis (LF) Non-governmental Development Organization (NGDO) Network was established to engage in supporting both international and national LF elimination agendas covering areas such as assisting ministries of health as an on-the-ground link between communities and programmes, which additionally gives the Network members an important voice from the field at international meetings; playing key roles in programme evolution (especially helping to both scale up and scale down mass drug administration [MDA] as elimination thresholds are met); having a role in operational research and developing new programme delivery models that can be taken to scale (such as linkages with other disease programmes and approaches to morbidity management and disability prevention); developing advocacy and policy approaches with other partners; convening other important stakeholders (academic, technical, programmatic and funding); mobilizing financial and technical resources to support programmes; supporting national human resource capacity building to catalyse national ownership of LF programmes; providing leadership in LF governance structures and working in areas of conflict to ensure that everybody in LF-endemic areas enjoys treatment services. Three case studies will illustrate the roles identified for NGDOs in LF programmes covering development of operational research, policy and advocacy linkage between LF and malaria programmes; launching LF morbidity management projects and NGDO\'s ability to work and deliver LF services in areas of conflict. In addition, the case studies will show the role of NGDOs in mobilising financial and technical resources that support national human resources, leading to national ownership of programmes. Conclusions will be drawn on the role of NGDOs in the Global Alliance for LF elimination and the need for continued partnerships to reach programme goals.

To eliminate lymphatic filariasis (LF) by 2020, the World Health Organization (WHO) has launched a campaign against the disease. Since the launch in 2000, significant progress has been made to achieve this ambitious goal. In this article we review the progress and status of the LF programme in Africa through the WHO neglected tropical diseases preventive chemotherapy databank, the Expanded Special Project for Elimination of Neglected Tropical Diseases (ESPEN) portal and other publications. In the African Region there are 35 countries endemic for LF. The Gambia was reclassified as not requiring preventive chemotherapy in 2015, while Togo and Malawi eliminated LF as a public health problem in 2017 and 2020, respectively. Cameroon discontinued mass drug administration (MDA) and transitioned to post-MDA surveillance to validate elimination. The trajectory of coverage continues to accelerate; treatment coverage increased from 0.1% in 2000 to 62.1% in 2018. Geographical coverage has also significantly increased, from 62.7% in 2015 to 78.5% in 2018. In 2019, 23 of 31 countries requiring MDA achieved 100% geographic coverage. Although much remains to be done, morbidity management and disability prevention services have steadily increased in recent years. Vector control interventions conducted by other programmes, particularly malaria vector control, have had a profound effect in stopping transmission in some endemic countries in the region. In conclusion, significant progress has been made in the LF programme in the region while we identify the key remaining challenges in achieving an Africa free of LF.

The discovery of naturally occurring B12-depedent photoreceptors has allowed for applications of cobalamins (Cbls) in optogenetics and synthetic biology to emerge. However, theoretical investigations of the complex mechanisms of these systems have been lacking. Adenosylcobalamin (AdoCbl)-dependent photoreceptor, CarH, is one example and it relies on daylight to perform its catalytic function. Typically, in enzymes employing AdoCbl as their cofactor, the Co-C5\' bond activation and cleavage is triggered by substrate binding. The cleavage of the Co-C5\' bond is homolytic resulting in radical pair formation. However, in CarH, this bond is instead activated by light. To explore this peculiarity, the ground and first excited state potential energy surfaces (PESs) were constructed using the quantum mechanics/molecular mechanics (QM/MM) framework and compared with other AdoCbl-dependent enzymes. QM/MM results indicate that CarH is photolytically active as a result of the AdoCbl dual role, acting as a radical generator and as a substrate. Photo-cleavage of the Co-C5\' bond and subsequent H-atom abstraction is possible because of the specific orientation of the H-C4\' bond with respect to the Co(II) center. Comparison with other AdoCbl-dependent enzymes indicate that the protein environment in the CarH active center alters the photochemistry of AdoCbl by controlling the stereochemistry of the ribose moiety.