Cholesterol-depleting statin drugs protect postmitotically differentiated human neurons against ethanol- and human immunodeficiency virus type 1-induced oxidative stress in vitro

The majority of human immunodeficiency virus type 1 (HIV-1)-infected individuals are either alcoholics or prone to alcoholism. Upon ingestion, alcohol is easily distributed into the various compartments of the body, particularly the brain, by crossing through the blood-brain barrier. Both HIV-1 and alcohol induce oxidative stress, which is considered a precursor for cytotoxic responses. Several reports have suggested that statins exert antioxidant as well as anti-inflammatory pleiotropic effects, besides their inherent cholesterol-depleting potentials. In our studies, postmitotically differentiated neurons were cocultured with HIV-1-infected monocytes, T cells, or their cellular supernatants in the presence of physiological concentrations of alcohol for 72 h. Parallel cultures were pretreated with statins (atorvastatin and simvastatin) with the appropriate controls, i.e., postmitotically differentiated neurons cocultured with uninfected cells and similar cultures treated with alcohol. The oxidative stress responses in the presence/absence of alcohol in these cultures were determined by the production of the well-characterized oxidative stress markers, 8-isoprostane-F2-alpha, total nitrates as an indicator for various isoforms of nitric oxide synthase activity, and heat shock protein 70 (Hsp70). An in vitro culture of postmitotically differentiated neurons with HIV-1-infected monocytes or T cells as well as supernatants from these cells enhanced the release of 8-isoprostane-F2-alpha in the conditioned medium six- to sevenfold (monocytes) and four- to fivefold (T cells). It was also observed that coculturing of HIV-1-infected primary monocytes over a time period of 72 h significantly elevated the release of Hsp70 compared with that of uninfected controls. Cellular supernatants of HIV-1-infected monocytes or T cells slightly increased Hsp70 levels compared to neurons cultured with uninfected monocytes or T-cell supernatants (controls). Ethanol (EtOH) presence further elevated Hsp70 in both infected and uninfected cultures. The amount of total nitrates was significantly elevated in the coculture system when both infected cells and EtOH were present. Surprisingly, pretreatment of postmitotic neurons with clinically available inhibitors of HMG-coenzyme A reductase (statins) inhibited HIV-1-induced release of stress/toxicity-associated parameters, i.e., Hsp70, isoprostanes, and total nitrates from HIV-1-infected cells. The results of this study provide new insights into HIV-1 neuropathogenesis aimed at the development of future HIV-1 therapeutics to eradicate viral reservoirs from the brain.     

Assessment of sorghum genetic resources of Ethiopia for anthracnose (Colletotrichum sublineolum Henn.) resistance and agronomic traits

Sorghum anthracnose caused by Colletotrichum sublineolum Henn. is one of the key diseases limiting sorghum production and productivity. Development of anthracnose‐resistant sorghum genotypes possessing yield‐promoting agronomic traits is an important breeding goal in sorghum improvement programs. The objective of this study was to determine the responses of diverse sorghum genetic resources for anthracnose resistance and agronomic traits to identify desirable lines for breeding. A total of 366 sorghum collections and three standard checks were field evaluated during the 2016 and 2017 cropping seasons. Lines were artificially inoculated with a virulent pure isolate of the pathogen. Anthracnose disease severity was assessed to calculate the area under disease progress curve (AUDPC). Agronomic traits such as panicle length (PL), panicle width (PW), head weight (HW) and thousand grain weight (TGW) were measured. Lines showed highly significant differences (p < .001) for anthracnose severity, AUDPC and agronomic traits. Among the collections 32 lines developed levels of disease severity between 15% and 30% in both seasons. The following sorghum landraces were selected: 71708, 210903, 74222, 73955, 74685, 74670, 74656, 74183, 234112, 69412, 226057, 214852, 71420, 71484, 200126, 71557, 75120, 71547, 220014, 228179, 16212, 16173, 16133, 69088, 238388, 16168 and 71570. These landraces had a relatively low anthracnose severity possessing farmer‐preferred agronomic traits. The selected genotypes are useful genetic resources to develop anthracnose‐resistant sorghum cultivars.     

Seasonal variation in primary production of a large high altitude tropical lake (Lake Tana,Ethiopia): effects of nutrient availability and water transparency

Primary production rates, chlorophyll and phytoplankton biovolume were measured monthly from April 2003 to November 2004 in Lake Tana, a large tropical lake in the highlands of Ethiopia. The lake is characterised by low nutrient concentrations, and a low water transparency due to high silt load of the inflowing rivers during the rainy seasons (May–November) and daily resuspension of sediments in the inshore zone. The mean chlorophyll-a concentrations varied seasonally and ranged from 2.6 mg m⁻³ to 8.5 mg m⁻³ (mean: 4.5 mg m⁻³) in the offshore zone. Primary production was measured using the light–dark bottles technique. We incubated only at three depths, i.e. 0.6, 1.2 and 1.8 m. Therefore, we may have missed a substantial part of the depth production profile and probably also frequently missed P _max. Gross primary production in the openwater averaged 2.43 g O₂ m⁻² d⁻¹ and ranged between 0.03 g O₂ m⁻² d⁻¹ and 10.2 g O₂ m⁻² d⁻¹; production was significantly higher in the inshore zone. The highest production rates were observed in the post-rainy season (Oct–Nov), which coincided with a bloom of Microcystis and higher chlorophyll levels. This seasonal high production is probably caused by a relatively high nutrient availability in combination with favourable light conditions. The gross primary production rates of L. Tana are among the lowest compared with other tropical lakes. This will be partly the result of our underestimation of gross primary production by often missing P _max. Another cause is the oligotrophic nature of the lake in combination with its relatively low water transparency. The gross primary production per unit chlorophyll in the openwater zone was in the same range as in 30 other tropical lakes and reservoirs. The higher primary production in the inshore zone is probably the result of the daily water column mixing (Z _mix ≥ Z _t) in this area, enhancing nutrient recycling. A large proportion of the annual primary production is realised in one of the four seasons only. This productive post-rainy season is relatively short (2 months) and therefore efficiency of transfer of matter between the first and second trophic level of the Lake ecosystem will be poor.     

The biogeochemistry of two forested catchments in the Black Forest and the eastern Ore Mountains (Germany)

The biogeochemical input-output fluxes of two forested catchments with contrasting levels of atmospheric deposition were investigated in Germany. This paper focuses on the effects of recent changes in atmospheric inputs on the chemical composition in the soil solution and stream. The catchment 'Schluchsee' (Black Forest; SW Germany) is characterized by relatively low atmospheric inputs whereas 'Rotherdbach' (Ore Mountains; E Germany) received significant amounts of acid deposition (mainly originating from SO₂ emissions) until recent years. Both sites reveal decreases in H⁺ and S deposition during the 1990s. This pattern is typical when compared to trends in Europe. In response to the reduced S deposition, soil solution and streamwater SO₄ ^2– concentrations decreased significantly. A net release of SO₄ ^2– (output > input) was observed at both sites due to the release of S previously stored in the soil. The level of N deposition was more or less constant at both sites. At Schluchsee, NO₃ ^– concentration in streamwater remained more or less unchanged, whereas a decrease at Rotherdbach was observed. A recovery from acidification was found in seepage water as indicated by increasing acid neutralizing capacity (ANC). Streamwater ANC increased only in the permanently acidified Rotherdbach. No change of ANC was observed in the Schluchsee stream, which was characterized by episodic acidification during high-flow conditions. Nevertheless, the key factor controlling the recovery from surface water acidification was the type, amount and distribution of stored S pools in the ecosystem. Thus, time series analysis of long-term data of input-output chemistry can be a valuable instrument in order to improve the understanding of linked terrestrial-aquatic systems and give useful clues for modeling efforts.     

Antigenic Properties of the HIV Envelope on Virions in Solution

The structural flexibility found in human immunodeficiency virus (HIV) envelope glycoproteins creates a complex relationship between antigenicity and sensitivity to antiviral antibodies. The study of this issue in the context of viral particles is particularly problematic as conventional virus capture approaches can perturb antigenicity profiles. Here, we employed a unique analytical system based on fluorescence correlation spectroscopy (FCS), which measures antibody-virion binding with all reactants continuously in solution. Panels of nine anti-envelope monoclonal antibodies (MAbs) and five virus types were used to connect antibody binding profiles with neutralizing activities. Anti-gp120 MAbs against the 2G12 or b12 epitope, which marks functional envelope structures, neutralized viruses expressing CCR5-tropic envelopes and exhibited efficient virion binding in solution. MAbs against CD4-induced (CD4i) epitopes considered hidden on functional envelope structures poorly bound these viruses and were not neutralizing. Anti-gp41 MAb 2F5 was neutralizing despite limited virion binding. Similar antigenicity patterns occurred on CXCR4-tropic viruses, except that anti-CD4i MAbs 17b and 19e were neutralizing despite little or no virion binding. Notably, anti-gp120 MAb PG9 and anti-gp41 MAb F240 bound to both CCR5-tropic and CXCR4-tropic viruses without exerting neutralizing activity. Differences in the virus production system altered the binding efficiencies of some antibodies but did not enhance antigenicity of aberrant gp120 structures. Of all viruses tested, only JRFL pseudoviruses showed a direct relationship between MAb binding efficiency and neutralizing potency. Collectively, these data indicate that the antigenic profiles of free HIV particles generally favor the exposure of functional over aberrant gp120 structures. However, the efficiency of virion-antibody interactions in solution inconsistently predicts neutralizing activity in vitro.     

Sodium stibogluconate (SSG) & paromomycin combination compared to SSG for visceral leishmaniasis in East Africa: a randomised controlled trial

Background

Alternative treatments for visceral leishmaniasis (VL) are required in East Africa. Paromomycin sulphate (PM) has been shown to be efficacious for VL treatment in India.

Methods

A multi-centre randomized-controlled trial (RCT) to compare efficacy and safety of PM (20 mg/kg/day for 21 days) and PM plus sodium stibogluconate (SSG) combination (PM, 15 mg/kg/day and SSG, 20 mg/kg/day for 17 days) with SSG (20 mg/kg/day for 30 days) for treatment of VL in East Africa. Patients aged 4–60 years with parasitologically confirmed VL were enrolled, excluding patients with contraindications. Primary and secondary efficacy outcomes were parasite clearance at 6-months follow-up and end of treatment, respectively. Safety was assessed mainly using adverse event (AE) data.

Findings

The PM versus SSG comparison enrolled 205 patients per arm with primary efficacy data available for 198 and 200 patients respectively. The SSG & PM versus SSG comparison enrolled 381 and 386 patients per arm respectively, with primary efficacy data available for 359 patients per arm. In Intention-to-Treat complete-case analyses, the efficacy of PM was significantly lower than SSG (84.3% versus 94.1%, difference = 9.7%, 95% confidence interval, CI: 3.6 to 15.7%, p = 0.002). The efficacy of SSG & PM was comparable to SSG (91.4% versus 93.9%, difference = 2.5%, 95% CI: −1.3 to 6.3%, p = 0.198). End of treatment efficacy results were very similar. There were no apparent differences in the safety profile of the three treatment regimens.

Conclusion

The 17 day SSG & PM combination treatment had a good safety profile and was similar in efficacy to the standard 30 day SSG treatment, suggesting suitability for VL treatment in East Africa.

Clinical Trials Registration

www.clinicaltrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00255567","term_id":"NCT00255567"}}NCT00255567     

Leishmania aethiopica: identification and characterization of cathepsin L-like cysteine protease genes

There is limited information on the biology and pathogenesis of Leishmania aethiopica, causative agent of cutaneous leishmaniasis (CL) in Ethiopia. In this study we have identified and characterized two cathepsin L-like cysteine protease genes, Laecpa and Laecpb, from L. aethiopica. The predicted amino acid sequence of Laecpa and Laecpb is more than 75% identical with homologous cathepsin L-like cysteine protease genes of other Leishmania species and less than 50% identical with human cathepsin L. Laecpa is expressed predominantly in the stationary, and to a lower level, during the amastigote stage while Laecpb is specifically expressed in the stationary stage of L. aethiopica development. Phylogenetic analysis showed that the two genes are grouped into separate clades which are the result of gene duplication. The isolation of these genes will be useful in developing Leishmania species specific diagnostics for molecular epidemiological studies and serves as a first step to study the role of cysteine proteases in L. aethiopica pathogenesis.     

Mechanisms of water economy in lactating Ethiopian Somali goats during repeated cycles of intermittent watering

We investigated the physiological and behavioural strategies by which lactating Ethiopian Somali goats endure repeated periods of water deprivation. The experiment lasted for 32 days and was divided into four periods of 8 days each. Measurements were taken during the first 4 days in each period. Seven does (W1) were watered once daily and seven does (W4) were watered once every 4th day. Rectal temperature was taken at 0800 and 1715 h. Blood samples were taken in the evening and milk samples in the morning. The goats were on pasture between 0900 to 1215 h and 1315 to 1630 h with behaviour recorded every 5 min. The does were supplemented with 300 g of concentrates per head per day. Plasma and milk osmolality were determined by freezing point depression. Plasma total protein was measured on a TS refractometer. Plasma vasopressin concentrations were analysed by radio-immunoassay. The mean daily water intake of W1 was 1897 ml compared with the calculated mean of 1075 ml in W4 (P < 0.001). The mean diurnal variation of the rectal temperature was 3.5°C in both groups. Afternoon rectal temperature in W4 during period 1 was higher than that in the W1 on the days of water deprivation (P < 0.05). With repeated periods, plasma osmolality in W4 increased less over the days of water deprivation. It was 336, 321, 311 and 306 mosm/l on the 4th day at periods 1, 2, 3 and 4, respectively. The corresponding vasopressin concentrations were 10.0, 9.2, 4.2 and 4.4 pmol/l. Total plasma protein concentration during period 1 on days 3 and 4 were higher in W4 than in W1 (P < 0.01). During the subsequent periods, it did not increase more in W4 compared with W1, but it was lower in W4 on the days after watering. W4 milk production decreased by 22% compared with W1 in all periods. With increasing days of water deprivation, the W4 goats spent less time in the sun, grazed shorter time and frequently ate cactus (Opuntia ficus-indica) as compared with W1. Results suggested that dehydrated lactating Ethiopian Somali goats economised on water by diurnal variations of rectal temperature, storing water in the extracellular fluid, by changing behaviour at grazing and by decreasing milk production.     

Clinico-mycological Profile of Dermatophytosis in a Reference Centre for Leprosy and Dermatological Diseases in Addis Ababa

To assess the clinical and fungal species spectrum of dermatophyte infection in a reference centre in Addis Ababa, 539 dermatological patients with signs of dermatophytosis were investigated. Seventy-one percent were female and 29% male, aged 2–66 years (median 9). Four hundred-fifteen (77%) had at least one skin lesion. Tinea capitis was diagnosed in 138/155 males (89%) as compared to 214/384 females (40%) (p<0.05). T. capitis was diagnosed in 69% of the 374 children. Fingernails were affected in 132/145 (91%) of onychomycosis, 118 (90%) of these patients were females and 14 males (p<0.05). Tinea corporis was observed in 45, and other types of tinea in 12 patients. Thirty-six percent of all patients had also other skin lesions, mostly impetigo. Of 490 cultured samples 364 (74%) grew dermatophytes: Trichophyton violaceum in 84%, Trichophyton verrucosum in 9.6%, Trichophyton tonsurans in 1.4% and T. rubrum in 0.5%. Additionally, 15 isolates were identified as white variants of T. violaceum, in 3 cases confirmed by sequencing of the rDNA ITS 2 region. T. capitis in young males and T. unguium of fingernails in females were the most common manifestations of dermatophytosis in Addis Ababa, usually caused by T.violaceum.