Moringa oleifera leaf fractions attenuated Naje haje venom-induced cellular dysfunctions via modulation of Nrf2 and inflammatory signalling pathways in rats

Naja haje envenoming could activate multiple pathways linked to haematotoxic, neurological, and antioxidant systems dysfunctions. Moringa oleifera has been used in the management of different snake venom-induced toxicities, but there is no scientific information on its antivenom effects against Naja haje. This study thus, investigated the antivenom activities of different extract partitions of M. oleifera leaves against N. haje envenoming. Forty five male rats were divided into nine groups (n = 5). Groups 2 to 9 were envenomed with 0.025 mg/kg (LD₅₀) of N. haje venom while group 1 was given saline. Group 2 was left untreated, while group 3 was treated with polyvalent antivenom, groups 4, 6 and 8 were treated with 300 mg/kg^?1 of N-hexane, ethylacetate and ethanol partitions of M. oleifera, respectively. Groups 5, 7 and 9 were also treated with 600 mgkg^?1of the partitions, respectively. Ethanol extract and ethyl acetate partition of M. oleifera significantly improved haematological indices following acute anaemia induced by the venom. Likewise, haemorrhagic, haemolytic and anti-coagulant activities of N. haje venom were best inhibited by ethanol partition. Envenoming significantly down-regulated Nuclear factor erythroid 2-related factor 2 (Nrf2) with the consequent elevation of antioxidant enzymes activities in the serum and brain. Treatment with extract partitions however, elevated Nrf2 levels while normalising antioxidant enzyme activities. Furthermore, there were reduction in levels of pro-inflammatory cytokines (TNF-α and interleukin-1β) in tissues of treated envenomed rats. This study concludes that ethanol partition of M. oleifera was most effective against N. haje venom and could be considered as a potential source for antivenom metabolites.     

Response of soil properties and maize yield to simulated erosion by artificial topsoil removal

The study was aimed at identifying the soil properties responsible for maize yield decline on eroded soils and at quantifying their relationship with yield. Topsoil was artificially removed to incremental depths of 0, 5, 10, 15 and 20 cm to simulate various degrees of erosion. Maize growth and yield were monitored on the plots and soil physical and chemical properties were determined after two years (4 seasons) of cultivation. Soil pH was significantly higher on the control plot and decreased with increased depth of topsoil removal. Bulk density (BD) increased with depth of topsoil removal from a mean value of 1.38 g cm⁻³ under control to 1.55 g cm⁻³ at 20 cm depth of removal, while cone index of penetrometer resistance (CI) correspondingly increased from 1.09 g cm⁻² to 1.37 g cm⁻². Maize yield significantly decreased in the first year from 3.2 t ha⁻¹ on the control plot to 0.12 t ha⁻¹ where 20 cm of topsoil was removed and correspondingly from 1.85 to 0.09 t ha⁻¹ in the second year of cropping. Maize yield decreased exponentially with increase in depth of topsoil removal (r ²=0.99, P<0.01) with an average of 55% yield loss on the removal of just 5 cm topsoil. Soil organic carbon (SOC), BD, CI, field capacity (FC), pH and exchangeable Mg²⁺ were significantly correlated to maize yield parameters. However, factor analysis showed that the combination of SOC and exchangeable Mg²⁺ with soil physical properties (BD, FC, CI and depth of topsoil removal) explained 99% of variation in maize grain yield. The need for conservation farm practices is recommended on the soil to prevent soil degradation. Section Editor: L. Wade     

Brief communication: Emergence chronology of permanent teeth in Nigerian children

This study measured the mean age, duration, and sequence of the emergence of permanent dentition in Nigerian children and compared the findings with other population groups. The cross‐sectional study involved 1,078 Nigerian children, aged 4–16 years old, from selected primary and secondary schools in the Ife Central local government area in Ile‐Ife, Osun State. In general, compared to boys, girls had an earlier mean age of emergence of all the permanent teeth. Children from high socioeconomic class had an earlier mean age of emergence for the maxillary incisors (6.43 and 7.58 years) and mandibular incisors (5.28 and 6.44 years) compared to children from middle and low socioeconomic classes, although socioeconomic effects were more mixed for premolars and molars. Compared to their counterparts in the USA, Australia, Belgium, and Iran, Nigerian children showed an earlier mean age of emergence of all the permanent teeth studied except for Pakistani boys, who had an earlier mean age of emergence of the maxillary premolars and second molar. Poorer economic status has been associated with delayed dental development; however, when compared to other populations, the Nigerian children in this study have earlier mean emergence ages than children from wealthier countries such as the USA and Australia. Am J Phys Anthropol 153:506–511, 2014. © 2013 Wiley Periodicals, Inc.     

Targeted Deletion of FGL2 Leads to Increased Early Viral Replication and Enhanced Adaptive Immunity in a Murine Model of Acute Viral Hepatitis Caused by LCMV WE

Mounting effective innate and adaptive immune responses are critical for viral clearance and the generation of long lasting immunity. It is known that production of inhibitory factors may result in the inability of the host to clear viruses, resulting in chronic viral persistence. Fibrinogen-like protein 2 (FGL2) has been identified as a novel effector molecule of CD4⁺CD25⁺ Foxp3⁺ regulatory T (Treg) cells that inhibits immune activity by binding to FCγRIIB expressed primarily on antigen presenting cells (APC). In this study, we show that infection of mice with Lymphocytic Choriomeningitis Virus WE (LCMV WE) leads to increased plasma levels of FGL2, which were detected as early as 2 days post-infection (pi) and persisted until day 50 pi. Mice deficient in FGL2 (fgl2^−/−) had increased viral titers of LCMV WE in the liver early p.i but cleared the virus by day 12 similar to wild type mice. Dendritic cells (DC) isolated from the spleens of LCMV WE infected fgl2^−/− had increased expression of the DC maturation markers CD80 and MHC Class II compared to wild type (fgl2^+/+). Frequencies of CD8⁺ and CD4⁺ T cells producing IFNγ in response to ex vivo peptide re-stimulation isolated from the spleen and lymph nodes were also increased in LCMV WE infected fgl2 ^−/− mice. Increased frequencies of CD8⁺ T cells specific for LCMV tetramers GP₃₃ and NP₃₉₆ were detected within the liver of fgl2^−/− mice. Plasma from fgl2^−/− mice contained higher titers of total and neutralizing anti-LCMV antibody. Enhanced anti-viral immunity in fgl2^−/− mice was associated with increased levels of serum alanine transaminase (ALT), hepatic necrosis and inflammation following LCMV WE infection. These data demonstrate that targeting FGL2 leads to early increased viral replication but enhanced anti-viral adaptive T & B cell responses. Targeting FGL2 may enhance the efficacy of current anti-viral therapies for hepatotropic viruses.     

Distribution of bacteria and fungi in the earthworm Libyodrillus violaceous (Annelida: Oligochaeta), a native earthworm from Nigeria

Earthworms are soil invertebrates that play a key role in recycling organic matter in soils. In Nigeria, earthworms include Libyodrillus violaceous. Aerobic and anaerobic bacterial counts, as well as fungal counts of viable microorganisms in soils and gut sections, were made on twenty L. violaceous collected from different sites on the campus of the University of Agriculture, Abeokuta, Nigeria. The samples were collected between April and November, 2002. Numbers of microorganisms were higher in castings and gut sections than in un-ingested soil samples. The guts and their contents also had higher moisture and total nitrogen contents than the un-ingested soils. Bacteria and fungi isolated from the samples were identified by standard microbiological procedures on the bases of their morphological and biochemical characteristics. Isolated bacteria were identified as Staphylococcus, Bacillus spp., Pseudomonas aeruginosa, Streptococcus mutans, Clostridium, Spirocheata spp., Azotobacter spp., Micrococcus lylae, Acinetobacter spp., Halobacterium for bacteria. Yeast isolates were identified as Candida spp., Zygosaccharomyces spp., Pichia spp., and Saccharomyces spp while molds were identified as, Aspergillus spp., Pytium spp., Penicillium spp., Fusarium spp and Rhizopus spp. Of the five locations examined, the refuse dump area had the highest numbers of both aerobic and anaerobic organisms, followed by the arboretum while the cultivated land area recorded the lowest counts. The higher numbers of microorganisms observed in the gut sections and casts of the earthworms examined in this work reinforce the general concept that the gut and casts of earthworms show higher microbial diversity and activity than the surrounding soil.     

Formulation of an effective mosquito-repellent topical product from Lemongrass oil

Ointment and cream formulations of lemongrass oil in different classes of base and the oil in liquid paraffin solution have been evaluated for mosquito repellency in a topical application. Mosquito repellency was tested by determining the bite-deterrence of product samples applied on an experimental bird's skin against a 2-day starved culture of Aedes aegypti L. mosquitoes. The 1%v/v solution and 15%v/w cream and ointment preparations of the oil exhibited > or =50% repellency lasting 2-3 h, which may be attributed to citral, a major oil constituent. This activity was comparable to that of a commercial mosquito repellent. Base properties of the lemongrass oil formulations influenced their effectiveness. The oil demonstrated efficacy from the different bases in the order of hydrophilic base > emulsion base > oleaginous base.     

Fine needle aspiration (FNA) in the management of palpable masses in Ibadan: impact on the cost of care

Fine needle aspiration cytology is a relatively new technique in the management of palpable lesions in Ibadan. In the University College Hospital (UCH) Ibadan, as in most centres in Nigeria, inadequate facilities, a heavy patient load, financial constraints and an unreliable supply of basic necessities like water, often delay definitive diagnosis and management. In order to alleviate the patient's problems and provide prompt and accurate diagnosis, an FNA Cytology Clinic was set up in the Pathology Department, UCH, Ibadan, managed by the pathology team. This report represents the results of our experience. The cost effectiveness and impact on cost of care are highlighted. FNA costs N250.00 (pounds sterling 2.00), whilst cost of biopsy can vary from N5000.00 to N10000.00 (pounds sterling 35.00 to pounds sterling 70.00). A total of 752 satisfactory smears was reviewed during the 3-year period 1995-97 from various sites including breast (n = 295), lymph node (n = 183) and thyroid (n = 143). Diagnostic accuracy varied with different sites, the accuracy rate for breast, lymph node and thyroid malignancy being approximately 100%, 80% and 93%, respectively.     

Human solid tumor xenografts in immunodeficient mice are vulnerable to lymphomagenesis associated with Epstein-Barr virus

Xenografting primary human solid tumor tissue into immunodeficient mice is a widely used tool in studies of human cancer biology; however, care must be taken to prove that the tumors obtained recapitulate parent tissue. We xenografted primary human hepatocellular carcinoma (HCC) tumor fragments or bulk tumor cell suspensions into immunodeficient mice. We unexpectedly observed that 11 of 21 xenografts generated from 16 independent patient samples resembled lymphoid neoplasms rather than HCC. Immunohistochemistry and flow cytometry analyses revealed that the lymphoid neoplasms were comprised of cells expressing human CD45 and CD19/20, consistent with human B lymphocytes. In situ hybridization was strongly positive for Epstein-Barr virus (EBV) encoded RNA. Genomic analysis revealed unique monoclonal or oligoclonal immunoglobulin heavy chain gene rearrangements in each B-cell neoplasm. These data demonstrate that the lymphoid neoplasms were EBV-associated human B-cell lymphomas. Analogous to EBV-associated lymphoproliferative disorders in immunocompromised humans, the human lymphomas in these HCC xenografts likely developed from reactivation of latent EBV in intratumoral passenger B lymphocytes following their xenotransplantation into immunodeficient recipient mice. Given the high prevalence of latent EBV infection in humans and the universal presence of B lymphocytes in solid tumors, this potentially confounding process represents an important pitfall of human solid tumor xenografting. This phenomenon can be recognized and avoided by routine phenotyping of primary tumors and xenografts with human leukocyte markers, and provides a compelling biological rationale for exclusion of these cells from human solid tumor xenotransplantation assays.     

Digit Sucking Habit and Association with Dental Caries and Oral Hygiene Status of Children Aged 6 Months to 12 Years Resident in Semi-Urban Nigeria

Objectives

Non-nutritive sucking (NNS) is a common behavior in childhood. The association between digit sucking, dental caries and oral health has been studied with inconclusive results. The objectives of this study were to determine the prevalence of, and the association between digit sucking, caries and oral hygiene status of children age six months to 12 years, resident in Ile-Ife, Osun State, Nigeria.

Methods

A cross-sectional study was conducted in Ife Central Local Government Area of Osun State. Data were collected through a household survey using a multi-stage sampling procedure from children between six months and 12 years. Details of each child’s socio-demographic characteristics, digit sucking habits, caries status and oral health status were collected. The association between digit sucking, caries status and oral hygiene status was determined using Chi square and Logistic regression.

Results

The mean age of the 992 study participants was 5.8 ± (3.2) years. The prevalence of digit sucking, caries and poor oral hygiene were 7.2%, 10.5% and 2.4% respectively. The mean dmft score was 0.22 ± (0.80), mean DMFT score was 0.04 ± (0.30) while mean Oral Hygiene Index score was 1.27 ± (0.73). Digit sucking increased the odds of having caries (OR: 1.28; CI: 0.58–2.81) but decreased the odds of having poor oral hygiene (OR: 0.58; CI: 0.34–1.01) insignificantly.

Conclusions

Digit sucking was not a significant predictor of caries and oral hygiene status, although the odds of having caries increased while the odds of having poor oral hygiene decreased with digit sucking.     

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

In vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate the human disease provide a valuable platform for research into disease pathophysiology and for the preclinical evaluation of novel therapies. We present a variety of methods to generate subcutaneous or orthotopic human HCC xenografts in immunodeficient mice that could be utilized in a variety of research applications. With a focus on the use of primary tumor tissue from patients undergoing surgical resection as a starting point, we describe the preparation of cell suspensions or tumor fragments for xenografting. We describe specific techniques to xenograft these tissues i) subcutaneously; or ii) intrahepatically, either by direct implantation of tumor cells or fragments into the liver, or indirectly by injection of cells into the mouse spleen. We also describe the use of partial resection of the native mouse liver at the time of xenografting as a strategy to induce a state of active liver regeneration in the recipient mouse that may facilitate the intrahepatic engraftment of primary human tumor cells. The expected results of these techniques are illustrated. The protocols described have been validated using primary human HCC samples and xenografts, which typically perform less robustly than the well-established human HCC cell lines that are widely used and frequently cited in the literature. In comparison with cell lines, we discuss factors which may contribute to the relatively low chance of primary HCC engraftment in xenotransplantation models and comment on technical issues that may influence the kinetics of xenograft growth. We also suggest methods that should be applied to ensure that xenografts obtained accurately resemble parent HCC tissues.