Isolation of chitinase gene induced during infection of vicia faba by botrytis fabae

The moderately resistant [Giza 716] and the susceptible [Giza 429] faba bean cultivars were used to identify some pathogenesis related proteins [PRs] associated with infection by chocolate spot disease. One isolate of Botrytis fabae purified from a plant sample taken from Nubaria location [Behera governorate, Egypt] was used in the artificial infection experiment. Qualitative and quantitative analyses were carried out on all protein banding patterns of the healthy and the infected faba bean leaves harvested at 8, 24 and 48 hr after inoculation. Data revealed that a 26 kDa protein band was more intensive 8, 24 and 48 hr after inoculation in cultivar Giza 716,. In addition, a 29 kDa protein band appeared after 24 and 48 hr. Furthermore, in cultivar Giza 429, 54 kDa protein bands appeared after 8, 24 and 48 hr post inoculation and 28 and 20 kDa appeared after 24 hr post inoculation.Reverse-Transcription [RT-PCR] showed that chitinase gene is expressed at very early stages in infected faba bean leaves. DNA fragment at molecular weight 900 bp appeared at 8, 24 and 48 hr after inoculation and disappeared in the healthy plants. The amplified products were cloned into pGEM-T Easy vector. Four clones named [PNAM1, PNAM2, PNAM3 and PNAM4] were selected for validation. The recombinant plasmids PNAM1, PNAM2 were verified for the presence of the Chitinase gene coding sequences by using both specific and universal primers in PCR. BNAM1-Chit-EG gene sequence showed 58.15% similarity when aligned with other Chitinase genes published in the gene bank

Body fat distribution, serum leptin, and insulin resistance in obese subjects with obstructive sleep Apnoea

Obstructive sleep apnoea [OSA] is strongly associated with obesity and is characterized by endocrine and metabolic changes. The aim of the present study is to clarify whether there is interrelationship between body fat, serum leptin, glucose-insulin metabolism and OSA

Subjects and measurements: We studied 23 obese subjects with OSA [13 males,and 10 females; age mean 36 +/- 4.4 years; BMI: 31.7 +/- 3.6 kg/m2; WHR: 1.2 +/- .25 in males and 0.81+.5 in females ;Apnoea Index "AI"[ 9.2 +/- 6.1] event/hour of sleep by means of overnight polysomnography; fasting glucose[109.8 +/- 21.4 mg/dL] ; fasting insulin[18.6 +/- 7.1 uU/L ]; IR[6.7 +/- 2.8]; fasting leptin[577.69 +/- 201.6 ng/ml]. Results were compared with those of 10 healthy normal weight subjects[6 males,4 females; age mean 36.8 +/- 4.4 years; BMI: 25 +/- 0.24 kg/m2; WHR: 0.86 +/- 0.01; AI: 2.1 +/- 1.1 event/hour; fasting glucose[71.7 +/- 2.8 mg /dL ]; fasting insulin[15.3 +/- . 48 uU/L ]; IR[4.6 +/- . 17]; fasting leptin [42.4 +/- 11.5 ng/ml]

Results: Anthropometric measurements of OSA subjects were highly significantly greater than controls; body weight [P<0.003]; BMI [P<0.00]; waist [P<0.000]; and WHR [P<0.000]. Fasting glucose levels; fasting plasma insulin; IR and leptin levels were significantly higher in OSA subjects than controls [P<0.000, 0.03, 0.002; and 0.000] respectively. Overnight polysomnography revealed significant difference between OSA subjects and controls as regards AI [P<0.001]. The major dependent outcome variable was the apnoea index [AI], "the average number of apnoeas per hour of sleep determined by overnight polysomnography". OSA was defined as AI >/= 5. Highly significant correlation between AI and WHR [P<0.00]; Fasting insulin [P<0.04]; IR [P<001] and Leptin [P<0.000] were detected. Also leptin concentrations correlated with fasting insulin [P<0.02]; IR [P<0.00] and WHR [P<0.000] besides the AI

In Conclusion: There is strong bidirectional, feed-forward pernicious correlation detected between OSA in one side and each of visceral obesity, leptin, and IR; also between leptin, obesity, and IR. This association may contribute to the pathological manifestations and somatic sequale of this condition. Leptin could have major role linking OSA with various metabolic abnormalities detected in obese subjects. High circulating leptin found in this study, suggests that both obesity and OSA may be caused by a leptin resistant state. Among obese subjects, it is visceral fat [WHR], rather than generalized obesity [BMI] that predisposes to OSA