Potential roles for eNOS and NrF2 /HO-1 signaling in the ameliorative effect of lixisenatide on diabetes-induced kidney injury in rats and its amplification by ticagrelor co-administration.

Antioxidant and anti-inflammatory effects of lixisenatide (LX) and ticagrelor (TC) have been previously identified in type 2 diabetes mellitus (T2DM). Diabetic nephropathy is one of the major complications of T2DM. In the current study, we examined the potential protective effects of LX and TC on experimentally induced diabetic nephropathy in T2DM rats and their possible molecular mechanisms. To examine this possibility, rats were fed a high-fat diet (HFD) for 12 weeks, followed by a single injection of 35 mg/kg streptozotocin (STZ) to induce T2DM. 10 µg/kg LX and 25 mg/kg TC were given alone or in combination to T2DM rats for 4 weeks. The kidney examination of T2DM rats showed clear deterioration. T2DM rats exhibited significantly higher body weight, blood glucose, hemostatic model assessment for insulin resistance (HOMA-IR), blood urea nitrogen (BUN), serum creatinine, kidney reactive oxygen species (ROS), nuclear factor-κ B (NF-κ B), and transforming growth factor-ß (TGF-ß ), and significantly lower serum insulin, urine creatinine, creatinine clearance (CRCL), kidney superoxide dismutase (SOD), glutathione reduced (GSH), nuclear factor erythroid 2 (NrF2 ), heme oxygenase-1 (HO-1), and endothelial nitric oxide synthase (eNOS) when compared to control rats. Single treatment with LX or TC showed obvious ameliorative effects on kidney complications in T2DM rats, with more ameliorative effects with the combined administration of both drugs. Conclusion: Our investigation found that both LX and TC could significantly ameliorate the development of diabetic nephropathy via stimulating NrF2 /HO-1 antioxidant pathway in addition to increasing eNOS and decreasing NF-κ B renal tissue concentrations, and these effects were markedly augmented by their combined administration.

Ameliorative effect of lixisenatide on diabetic cardiovascular damage and its enhancement via ticagrelor co-administration in rats: possible role of eNOS and NrF2 /HO-1 signaling.

In this study, we hypothesized that lixisenatide (LIX) and ticagrelor (TIC) could have a protective effect against type 2 diabetes mellitus (T2DM)-induced vascular damage. Furthermore, we explored the possible additional protective effect of co-administering LIX and TIC in the treatment regimen. Methods: 50 male rats were divided into five groups, each comprising 10 rats: C (control), D (T2DM rats), D + LIX (T2DM rats treated with LIX for 4 weeks), D + TIC (T2DM rats treated with TIC for 4 weeks), and D + LIX + TIC (T2DM rats treated with LIX + TIC for 4 weeks). Results: The D group showed an increase in body weight, blood glucose, hemostatic model assessment for insulin resistance (HOMA-IR), aorta reactive oxygen species (ROS), and nuclear factor kappa B (NF-κ B), along with a reduction in serum insulin, aorta superoxide dismutase (SOD), glutathione reduced (GSH), nuclear factor erythroid-2 (NrF2), hemeoxygenase-1 (HO-1), and endothelial nitric oxide synthase (eNOS). Deterioration in the aorta histopathological condition, coupled with a noticeable impairment in vascular reactivity compared to the C group, was observed. A single administration of LIX showed a reduction in body weight, blood glucose, HOMA-IR, aorta ROS, and NF-κ B, accompanied by an increase in serum insulin, aorta SOD, GSH, NrF2, HO-1, and eNOS. Amelioration in the aorta histopathological condition and improved vascular reactivity compared to the D group were reported. Similarly, a single administration of TIC showed a reduction in aorta ROS and NF-κ B, along with an increase in aorta SOD, GSH, NrF2, HO-1, and eNOS. A slight amelioration was detected in the aorta histopathological condition, with improved vascular reactivity compared to the D group. The combined administration of LIX and TIC showed a reduction in aorta ROS and NF-κ B, along with an increase in aorta GSH, SOD, HO-1, and eNOS. This was combined with evident amelioration in the aorta histopathological condition and noticeable improvement in vascular reactivity compared to the single treatment with either LIX or TIC group. Conclusion: The present study introduces clear evidence that the administration of LIX and TIC can improve metabolic and vascular complications of T2DM through modulating eNOS and NrF2 /HO-1 signaling. The combined administration of LIX and TIC produced more significant effects than a single treatment.

A large outbreak of gastrointestinal illness at an open-water swimming event in the River Thames, London.

Open-water swimming is increasingly popular, often in water not considered safe for bathing. Limited evidence exists on the associated health risks. We investigated gastrointestinal illness in 1100 swimmers in a River Thames event in London, UK, to describe the outbreak and identify risk factors. We conducted a retrospective cohort study. Our case definition was swimmers with any: diarrhoea, vomiting, abdominal cramps lasting ⩾48 h, nausea lasting ⩾48 h, with onset within 9 days after the event. We used an online survey to collect information on symptoms, demographics, pre- and post-swim behaviours and open-water experience. We tested associations using robust Poisson regression. We followed up case microbiological results. Survey response was 61%, and attack rate 53% (338 cases). Median incubation period was 34 h and median symptom duration 4 days. Five cases had confirmed microbiological diagnoses (four Giardia, one Cryptosporidium). Wearing a wetsuit [adjusted relative risk (aRR) 6·96, 95% confidence interval (CI) 1·04-46·72] and swallowing water (aRR 1·42, 95% CI 1·03-1·97) were risk factors. Recent river-swimming (aRR 0·78, 95% CI 0·67-0·92) and age >40 years (aRR 0·83, 95% CI 0·70-0·98) were protective. Action to reduce risk of illness in future events is recommended, including clarification of oversight arrangements for future swims to ensure appropriate risk assessment and advice is provided.

Interim report: review of evidence of the health impact of famine in Ethiopia.

Historical accounts of famines in Ethiopia go as far back as the 9th century, however, evidence on its impact on health only started to emerge from the 15th century onwards. Unfortunately, famine has been endemic in Ethiopia in the last few decades. The 1973 famine is reported to have claimed over 300,000 lives. In 1985 approximately 10 million people were reported to be starving, with approximately 300,000 already dead and about 1000 dying daily. In the following years, droughts leading to food shortage have had local and national adverse health effects, in particular in 1999/2000. This paper describes the initial findings of a literature review of evidence on the health impact of droughts leading to famine in Ethiopia and highlights gaps in knowledge. The key finding, thus far, is the marked paucity of health impact data. This review also highlights the fact that adverse health impacts of famines are probably complex and long lasting. Interpretation of any health impact data is difficult as there are few baseline data to compare. Health effects also impact livelihoods. Livelihood disruption following famine does not just affect one generation but also subsequent generations. Surveillance systems are needed so that records of the health impacts of a drought that leads to famine can inform action. With climate change bringing increased likelihood of drought and famine in some parts of the world, the findings of this review could be beneficial not just for Ethiopia but also elsewhere.

Seasonal and altitudinal variations in fly density and their association with the occurrence of trachoma, in the Gurage zone of central Ethiopia.

In the Gurage zone of central Ethiopia, the association between fly density and the occurrence of trachoma has been investigated across varying altitudes. The seasonal pattern of fly density in the area was also explored. When, over short sampling periods (10 min/child indoors and 10 min/child outdoors), hand nets were used to collect flies from the eyes of children aged 2-8 years, only Musca sorbens and M. domestica were caught. Almost all of the 13,147 'eye-seeking' flies collected came from villages at low (<2000 m; 40.7%) or mid altitudes (2200-2500 m; 58.6%) with only 0.7% of them caught in the high-altitude villages investigated (at >3000 m). Musca sorbens predominated outdoors and M. domestica indoors. Almost all (99.3%) of the eye-seeking M. sorbens collected were caught outdoors whereas most (76.7%) of the M. domestica were caught indoors (P<0.0001 for each). The median numbers of flies caught, per child, per 10-min collection, in the low-, mid- and high-altitude villages were 9.5, six and zero, respectively, for M. sorbens, and eight, three and zero, respectively, for M. domestica. The altitudinal trends in these numbers of 'eye-seeking' flies matched those in the prevalences of active trachoma among children aged 1-10 years, which were high in the villages at low (81.6%) and mid altitude (78.7%) but much lower (1.7%) in the high-altitude villages. In conclusion, trachoma is a common disease of public-health importance only in the low- and mid-altitude villages in the Gurage zone, where there are large numbers of eye-seeking flies, and not in the villages that lie >3000 m above sea level, where there is a dearth of such flies.

Production of recombinant protein Pap31 and its application for the diagnosis of Bartonella bacilliformis infection.

Tropical bartonellosis is a highly fatal epidemic and endemic infectious disease that occurs throughout the communities of the Andes Mountains in South America. The disease is caused by the facultative intracellular bacteria, Bartonella bacilliformis. The emergence of bartonellosis in new geographic areas and an increase in the number of reported cases suggest the need for a rapid test for epidemiologic study and investigation of the disease burden. The objective of this research is to develop a rapid serologic diagnostic test using recombinant antigens to overcome the limitations of the current standard IFA technique for laboratory diagnosis. Western blot analysis with patient sera of whole cell lysate separated on a 2D gel identified Pap31 as a dominant antigen. PCR primers were designed according to the sequence of ATCC strain 35685 to amplify the gene coding for Pap31 from a local isolate (HOSP 800-09, Peru). The amplicon was subsequently cloned into pET24a, adding the T7 tag, and expressed in E. coli. Patient sera with different IFA titers confirmed the diagnostic band of 31 kDa on a Western blot of SDS-PAGE. The performance of affinity-purified recombinant Pap31 (rPap31) was also evaluated in an ELISA format with 137 patient sera of known IFA titers. The range of ELISA reading from positive sera did not overlap with the range of those from negative sera, suggesting the potential application of rPap31 in both ELISA for high throughput regional hospital settings and in the construction of handheld rapid tests for rural clinical sites.

Histone deacetylase inhibitors reduce polyglutamine toxicity.

Polyglutamine diseases include at least nine neurodegenerative disorders, each caused by a CAG repeat expansion in a different gene. Accumulation of mutant polyglutamine-containing proteins occurs in patients, and evidence from cell culture and animal experiments suggests the nucleus as a site of pathogenesis. To understand the consequences of nuclear accumulation, we created a cell culture system with nuclear-targeted polyglutamine. In our system, cell death can be mitigated by overexpression of full-length cAMP response element binding protein (CREB)-binding protein (CBP) or its amino-terminal portion alone. CBP is one of several histone acetyltransferases sequestered by polyglutamine inclusions. We found histone acetylation to be reduced in cells expressing mutant polyglutamine. Reversal of this hypoacetylation, which can be achieved either by overexpression of CBP or its amino terminus or by treatment with deacetylase inhibitors, reduced cell loss. These findings suggest that nuclear accumulation of polyglutamine can lead to altered protein acetylation in neurons and indicate a novel therapeutic strategy for polyglutamine disease.

Codon optimization of gene fragments encoding Plasmodium falciparum merzoite proteins enhances DNA vaccine protein expression and immunogenicity in mice.

In contrast to conventional vaccines, DNA and other subunit vaccines exclusively utilize host cell molecules for transcription and translation of proteins. The adenine plus thymine content of Plasmodium falciparum gene sequences (approximately 80%) is much greater than that of Homo sapiens (approximately 59%); consequently, codon usage is markedly different. We hypothesized that modifying codon usage of P. falciparum genes encoded by DNA vaccines from that used by the parasite to those resembling mammalian codon usage would lead to increased P. falciparum protein expression in vitro in mouse cells and increased antibody responses in DNA-vaccinated mice. We synthesized gene fragments encoding the receptor-binding domain of the 175-kDa P. falciparum erythrocyte-binding protein (EBA-175 region II) and the 42-kDa C-terminal processed fragment of the P. falciparum merozoite surface protein 1 (MSP-1(42)) using the most frequently occurring codon in mammals to code for each amino acid, and inserted the synthetic genes in DNA vaccine plasmids. In in vitro transient-expression assays, plasmids containing codon-optimized synthetic gene fragments (pS plasmids) showed greater than fourfold increased protein expression in mouse cells compared to those containing native gene fragments (pN plasmids). In mice immunized with 0.5, 5.0, or 50 microg of the DNA plasmids, the dose of DNA required to induce equivalent antibody titers was 10- to 100-fold lower for pS than for pN plasmids. These data demonstrate that optimizing codon usage in DNA vaccines can improve protein expression and consequently the immunogenicity of gene fragments in DNA vaccines for organisms whose codon usage differs substantially from that of mammals.

CREB-binding protein sequestration by expanded polyglutamine.

Spinal and bulbar muscular atrophy (SBMA) is one of eight inherited neurodegenerative diseases known to be caused by CAG repeat expansion. The expansion results in an expanded polyglutamine tract, which likely confers a novel, toxic function to the affected protein. Cell culture and transgenic mouse studies have implicated the nucleus as a site for pathogenesis, suggesting that a critical nuclear factor or process is disrupted by the polyglutamine expansion. In this report we present evidence that CREB-binding protein (CBP), a transcriptional co-activator that orchestrates nuclear response to a variety of cell signaling cascades, is incorporated into nuclear inclusions formed by polyglutamine-containing proteins in cultured cells, transgenic mice and tissue from patients with SBMA. We also show CBP incorporation into nuclear inclusions formed in a cell culture model of another polyglutamine disease, spinocerebellar ataxia type 3. We present evidence that soluble levels of CBP are reduced in cells expressing expanded polyglutamine despite increased levels of CBP mRNA. Finally, we demonstrate that over-expression of CBP rescues cells from polyglutamine-mediated toxicity in neuronal cell culture. These data support a CBP-sequestration model of polyglutamine expansion disease.

Onchocerciasis in Gilgel Ghibe River Valley southwest Ethiopia.

OBJECTIVE: To determine the prevalence of onchocerciasis and the entomological transmission indices such as the parous rate and annual transmission potential (ATP). SETTING: Gilgel Ghibe village, Gilgel Ghibe River Valley Southwest Ethiopia between April 1994 and March 1995. SUBJECTS: Two hundred twenty eight subjects of the total 400 population in Gilgel Ghibe village were subjected to parasitological and clinical examinations. METHOD: Two skin snips per person were taken and examined for microfilariae of Onchocerca volvulus. Fly collections were done from dawn to dusk from human baits seated in pairs at four representative sites at the river bank and away from the river bank. Flies were dissected for parity and infections with O. volvulus larvae. RESULTS: Among the 228 people examined, the prevalence of the disease was low (17%), being higher in males (19%) than in females (14%). The geometric mean of microfilarial density was 11.1 (range, 1-132) mf per skin snip. Itching followed by pigmentary changes were the most common clinical signs and symptoms. The predominant anthropophilic blackfly species was Simulium (Edwardsellum) damnosum s.l. The annual parous rate and ATP were 74.7% and 1669.5, respectively, being higher at the river bank than at sites further away suggesting a greater risk of infection by the river side. CONCLUSION: The low prevalence of onchocerciasis in Gilgel Ghibe area vis-a-vis the high ATP level could be due to the possible presence of bovine onchocerciasis in the area. Further studies employing molecular techniques are thus required to identify O. volvulus from other filariae in flies.

PIP: 400 people in Gilgel Ghibe, southwestern Ethiopia, were subjected to parasitological and clinical examination to determine the prevalence and intensity of onchocerciasis. Its association with entomological transmission indices such as the parous rate and annual transmission potential (ATP) were determined simultaneously. Two skin snips per person were taken and examined for microfilariae of Onchocerca volvulus. In addition, collections of adult blackfly were done from human baits seated in pairs at 4 representative sites at the riverbank and away from the riverbank. Flies were then dissected for parity and infections with O. volvulus. Among the 228 people examined, the prevalence of the disease was low (17%), being higher in males than in females (19% vs. 14%). The geometric mean of microfilarial density was 11.1 mf per skin snip. Itching and skin changes were the most common signs and symptoms of the disease. The predominant anthropophilic blackfly species was Simulium (Edwardsellum) damnosum s.l. The annual parous rate was 74.7%, while ATP was 1669.5, being higher at the riverbank than at farther sites, suggesting a greater risk of infection by the riverside. In conclusion, the low prevalence of onchocerciasis vis-a-vis the high ATP level could be caused by the possible presence of bovine onchocerciasis in the area. Further studies employing molecular techniques are needed to identify O. volvulus from other filariae in flies.

Cleavage, aggregation and toxicity of the expanded androgen receptor in spinal and bulbar muscular atrophy.

Spinal and bulbar muscular atrophy (SBMA) is a neurodegenerative disease caused by the expansion of a polyglutamine repeat within the androgen receptor (AR). We have studied the mutant AR in an in vitro system, and find both aggregation and proteolytic processing of the AR protein to occur in a polyglutamine repeat length-dependent manner. In addition, we find the aberrant metabolism of expanded repeat AR to be coupled to cellular toxicity, indicating a likely molecular basis for the toxic gain of AR function that produces neuronal degeneration in SBMA.

Fine needle aspiration cytology of breast lesions: a clinicocytologic review of 1211 cases.

Fine needle aspiration (FNA) is a widely accepted cytologic technique for the early diagnosis of palpable breast lesions. Early diagnosis and treatment of benign and malignant breast lesions in turn are associated with increased chance of long term survival. Clinical data and results of fine needle aspiration done at the Department of Pathology in Tikur Anbessa Hospital between January 1991 to December 1994 were reviewed to study the cytologic features of breast lesions and to elucidate the clinical use of fine needle aspiration. A total of 9,946 fine needle aspirations were done in the department, out of which 1,211 (12.2%) were from breast lesions. Results from breast lesions were reported as: malignant, benign suspicious and unsatisfactory. Repeat aspiration was performed for all unsatisfactory cases. Fine needle aspiration of the 1,211 breast lesions in the four year period revealed 255 (21%) malignant, 901 (74%) benign, 15 (1.2%) suspicious and 40 (3.3%) unsatisfactory cases. The malignant lesions ranged from small, mobile, firm nodules measuring 4 cm to fixed ulcerated and fungating mass, measuring 20 cm or more. The benign lesions were small and mobile, measuring from 2 cm to 6 cm, mostly fibroadenoma (38%). Biopsy result was found for 20 patients. Out of these 14 were benign lesions and 6 were malignant lesions, confirmed by surgical biopsy. The mean age of patients with malignant lesions was 43 years (range: 24 to 80 years), while it was 27 years (range: 15 to 50 years) for those with benign lesion. There were 1132 (93.5%) females and 79 (6.5%) males. Fine needle aspiration diagnostic technique is recommended to be widely introduced into the health system of Ethiopia in order to ensure timely diagnosis of benign and malignant breast lesions. Furthermore a properly designed study will be beneficial to elucidate the cytologic and clinical characteristics of breast lesions in Ethiopian setting.