The promoter of a novel human papillomavirus (HPV77) associated with skin cancer displays UV responsiveness, which is mediated through a consensus p53 binding sequence.

An aetiological role has been proposed for human papillomavirus (HPV) in skin carcinogenesis within the immunosuppressed patient population. To examine this possibility, we have focused on an HPV type that, to date, has been identified only in the cutaneous lesions of renal transplant recipients despite a high degree of sequence homology with other HPVs commonly found in warts in the general population. We report that the non-coding region of this virus, HPV type 77, contains a consensus binding site for the tumour suppressor protein p53, and we show by gel-retardation analysis that this sequence does indeed bind p53. Furthermore, using reporter gene assays, we demonstrate that HPV77 promoter activity is stimulated by UV radiation and that this response is mediated through the p53 binding site. This is the first report of a p53-dependent positive response element within a viral genome. Our results suggest a possible novel mechanism by which specific types of HPV might act as cofactors with UV radiation in cutaneous transformation.     

Optimization of methods for peripheral blood mononuclear cells isolation and expansion of human gamma delta T cells

Human Vg9/Vδ2 T cells (γδ T cells) are immune surveillance cells both in innate and adaptive immunity and are a possible target for anticancer therapies, which can induce immune responses in a variety of cancers. Small non-peptide antigens such as zoledronate can do activation and expansion of T cells in vitro. It is evident that for adoptive cancer therapies, large numbers of functional cells are needed into cancer patients. Hence, optimization of methods needs to be carried out for the efficient expansion of these T cells. Standardization of peripheral blood mononuclear cells (PBMCs) isolation was devised. Cytokines (interleukin 2 (IL-2) and interleukin 15 (IL-15)) and zoledronate were also standardized for different concentrations. It was found that an increased number of PBMCs were recovered when washing was done at 1100 revolution per minute (rpm). Significantly high expansion fold was (2524 ± 787 expansion fold) achieved when stimulation of PBMCs was done with 1 µM of zoledronate and both cytokines IL-2 and IL-15 supported the expansion and survival of cells at the concentrations of 100 IU/ml and 10 ng/ml respectively. 14-day cultures showed highly pure (91.6 ± 5.1%) and live (96.5 ± 2.5%) expanded γδ T cells. This study aimed to standardize an easy to manipulate technique for the expansion of γδ T cells, giving a higher yield.     

An improved intrafemoral injection with minimized leakage as an orthotopic mouse model of osteosarcoma

Osteosarcoma, the most common type of primary bone cancer, is the second highest cause of cancer-related death in pediatric patients. To understand the mechanisms behind osteosarcoma progression and to discover novel therapeutic strategies for this disease, a reliable and appropriate mouse model is essential. For this purpose, osteosarcoma cells need to be injected into the bone marrow. Previously, the intratibial and intrafemoral injection methods were reported; however, the major drawback of these methods is the potential leakage of tumor cells from the injection site during or after these procedures. To overcome this, we have established an improved method to minimize leakage in an orthotopic mouse model of osteosarcoma. By taking advantage of the anatomical benefits of the femur with less bowing and larger medullary cavity than those of the tibia, osteosarcoma cells are injected directly into the femoral cavity following reaming of its intramedullary space. To prevent potential leakage of tumor cells during and after the surgery, the injection site is sealed with bone wax. This method requires a minor surgery of approximately 15 min under anesthesia. Our established orthotopic osteosarcoma model could serve as a valuable and reliable tool for examining progression of various types of bone tumors.     

The distribution and comparison of glutathione, glutathione reductase and glutathione S-transferase in various camel tissues

Extracts prepared from liver, kidney, lung and brain of camel contain glutathione, glutathione S-transferase and glutathione reductase. Liver had the highest level of glutathione (218.7 mumol/g wet weight) whereas brain had the lowest level (66.4 mumol/g wet weight). The highest activity for glutathione reductase was found in the kidney (2.6 mumol/min/mg protein) while the lowest activity was found in the lung (0.9 mumol/min/mg protein). Glutathione S-transferase activity was the highest in liver (4.2 mumol/min/mg protein) and the lowest in brain (1 mumol/min/mg protein). Purified glutathione S-transferases from lung, kidney, brain and liver were similar in their molecular size, subunit composition as well as immuno-reactivity and showed some differences in their response to heat and inhibitors.     

Guillain-Barré syndrome as a cause of acute flaccid paralysis in Iraqi children: a result of 15 years of nation-wide study

Background

Guillain-Barré syndrome (GBS) is the most common cause of acute flaccid paralysis (AFP) in the post-poliomyelitis eradication era. This is the first study done to identify the epidemiology, clinical features, and outcome of GBS in Iraqi children over 15 years.

Methods

The surveillance database about AFP cases?<?15 years reported during January 1997-December 2011 was used.

Results

GBS represented 52.5% of AFP cases, with an incidence of 1.33 case/100,000 population?<?15 years/year. There was a higher incidence in the Southern provinces, age group 1–4 years, males, and outside the capital city of province, with no significant seasonal variations (p?=?.22). Survival probability after the 1 year of onset for those with respiratory muscle involvement was .76 (95% CI: .60-.86), versus .97 (95% Cl: .96-.98) for those who did not develop it (p?<?.001); and .97 (95% CI: .96-.98) for those living inside the capital city, versus .94 (.93-.95) for those living outside (p?=?.001). Cumulative incidence of residual paralysis for patients living inside the capital city was .21 (95% CI: .18-.24), versus .27 (95% CI: .25-.29) for those living outside (p?<?.001).

Conclusions

The incidence, age and gender distribution, and seasonality of GBS among Iraqi children is similar to those reported from other previous studies. It is the most important cause of AFP, especially in those between the age of 1 to 4 years living in rural areas.

     

A histological,immunohistochemical and biochemical study of the effects of pomegranate peel extracts on gibberellic acid induced oxidative stress in adult rat testes

ABSTRACT

Gibberellins are commonly used plant growth regulators that exhibit deleterious effects on various animal tissues. We investigated the histological, immunohistochemical and biochemical effects of gibberellic acid (GA3) on rat testes as well as the possible protective role of pomegranate peel extract (PPE). We used 28 adult male rats divided into control, PPE treated, GS3 treated and GA3 + PPE treated groups. Testis specimens were analyzed for superoxide dismutase (SOD) and catalase (CAT) activity, and examined histologically. We also investigated the androgen receptor using immunohistochemistry. The GA3 treated group exhibited significantly decreased SOD and CAT levels and area percent of androgen receptor. Seminiferous tubules (ST) were widely separated and the germinal epithelium was separated from the basement membrane in some tubules. Areas of vacuolation, degenerated germ cells with pyknotic nuclei and large multinucleated cells were observed. Ultrastructurally, primary spermatocytes exhibited vacuolated cytoplasm, degenerated mitochondria and hyperchromatic nuclei. Degenerated early spermatids with a ruptured or hyperchromatic nucleus were found. Spermatozoa exhibited abnormalities of the head and tail. The cytoplasm of Sertoli and Leydig cells exhibited dilated smooth endoplasmic reticulum. A significant improvement of the biochemical, histological and immunohistochemical alterations was observed in the GA3 + PPE treated group compared to the GA3 treated group.     

Investigation of some physical,chemical, and bacteriological parameters of water quality in some dams in Albaha region,Saudi Arabia

This study was conducted to evaluate the quality of water in selected dams in Albaha region, Kingdom of Saudi Arabia. Water samples from eight dams were subjected to physical, chemical, and bacteriological assessment using standardized procedures of conductivity, total dissolved solids, ions, acidity & alkalinity, and EC blue 100® coliform detection. About three fourth (75%) of dams’ water samples exceeded the permissible levels of pH, total dissolved solids, turbidity, Mn and NO₃ set by Saudi standards. Average levels of total dissolved solids, Fe, Mn, SO₄, NO₃, and NO₂ were 3065.00, 0.10, 0.89, 68.25, 17.91 and 0.016 mg/L, respectively. However, the average pH of water samples was 7.95 ± 0.66 which still within the accepted range set by national and global standards. Moreover, total dissolved solids also exceeded regular standards of Food and Agriculture Organization for irrigation water quality. Coliform bacteria were detected in 37.5% of dams without any significant spatial differences between dams and sites as groups. Correlations were found between pH & NO₃, SO₄ & NO₃, coliform bacteria & turbidity, coliform bacteria & NO₂ levels. Increased concentrations of assessed parameters in dams may be attributed to agricultural activities as well as animal and human wastes deposited into dams via rainfalls and flash floods. Proper treatment of dams needs to be taken into account before consumption and irrigation.