Conserved amino acid residues that are important for ligand binding in the type I gonadotropin-releasing hormone (GnRH) receptor are required for high potency of GnRH II at the type II GnRH receptor

GnRH I regulates reproduction. A second form, designated GnRH II, selectively binds type II GnRH receptors. Amino acids of the type I GnRH receptor required for binding of GnRH I (Asp2.61(98), Asn2.65(102), and Lys3.32(121)) are conserved in the type II GnRH receptor, but their roles in receptor function are unknown. We have delineated their functions using mutagenesis, signaling and binding assays, immunoblotting, and computational modeling. Mutating Asp2.61(97) to Glu or Ala, Asn2.65(101) to Ala, or Lys3.32(120) to Gln decreased potency of GnRH II-stimulated inositol phosphate production. Consistent with proposed roles in ligand recognition, mutations eliminated measurable binding of GnRH II, whereas expression of mutant receptors was not decreased. In detailed analysis of how these residues affect ligand-dependent signaling, [Trp2]-GnRH I showed lesser decreases in potency than GnRH I at the Asp2.61(97)Glu mutant. In contrast, [Trp2]-GnRH II showed the same loss of potency as GnRH II at this mutant. This suggests that Asp2.61(97) contributes to recognition of His2 of GnRH I, but not of GnRH II. GnRH II showed a large decrease in potency at the Asn2.65(101)Ala mutant compared with analogs lacking the CO group of Gly10NH2. This suggests that Asn2.65(101) recognizes Gly10NH2 of GnRH II. GnRH agonists showed large decreases in potency at the Lys3.32(120)Gln mutant, but antagonist activity was unaffected. This suggests that Lys3.32(120) recognizes agonists, but not antagonists, as in the type I receptor. These data indicate that roles of conserved residues are similar, but not identical, in the type I and II GnRH receptors.     

Evaluation of bioactive compounds,free radical scavenging and anticancer activities of bulb extracts of Boophone disticha from Eastern Cape Province,South Africa

Boophone disticha (B. disticha) is a bulbous tropical and subtropical flowering plant widespread in Africa, which is frequently used to treat several human ailments. Until the present, there is no scientific validation on the biological activity of this plant from the Eastern Cape Province of South Africa and as a result, this study aimed to assess the bioactive compounds, free radicals scavenging and anticancer potentials of crude bulb extracts (chloroform, acetone, and ethanol) of Boophone disticha obtained from this geographical location. Standard biochemical techniques and Gas-chromatography mass spectrometry (GCMS) analysis were used to pinpoint the bioactive compounds in the crude extracts sequel to their antioxidant potentials against radicals such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), hydrogen peroxide and nitric oxide as well as their ferric ion reducing power. In addition, their cytotoxicity effects against Human cervix adenocarcinoma (HeLa) cells were assessed as an in vitro model for anticancer. The phytochemical evaluation of the crude extracts showed the presence of phenolics, flavonoids, and alkaloids. GCMS profiles confirmed the presence of some bioactive compounds in the crude extracts of B. disticha that could be responsible for their biological activities. The plant extracts possessed considerable antioxidant activity and exhibited dose-dependent radicals’ inhibition from all assays carried out. Furthermore, the cytotoxicity effects against HeLa cells recorded inhibition concentration (IC₅₀) of 1.5, 1.6, and 1.9 µg/mL for acetone, chloroform, and ethanolic extracts of B. disticha, respectively. Findings from the present study suggest that B. disticha could be a good prospective source of antioxidant and anticancer agents. Therefore, further research on the isolation and purification of compounds from these extracts are indispensable.     

Relationship between acid phosphatase activity and P concentration in organs of Cyclopia and Aspalathus species,and a non-legume of the Cape Floristic Region

Aims The role of tissue acid phosphatase (APase) activity of legumes and non-legumes in their P nutrition and adaptation to low-P soils is not well understood. To better understand this, a relationship between APase activity and P concentration in leaves, stems, roots and nodules of legumes,CyclopiaandAspalathusand a non-legume,Leucadendron strictum, all native to the P-poor soils of the Cape fynbos biome, was assessed.     

Nitrogen nutrition,carbon accumulation and δ13C of Cyclopia and Aspalathus species in different settings of the Cape fynbos,South Africa

Aims Cyclopia and Aspalathus are legumes harvested for production of Honeybush and Rooibos tea, respectively. Farmers grow these species from either seeds or cuttings over several years with continuous annual harvesting. The aims of this study were to assess the effect of plant age, plant species, toposequence, planting material and farmer practice on nitrogen (N) nutrition and water-use efficiency of two Cyclopia and Aspalathus species in the Cape fynbos.Methods The study was conducted using plants from Koksrivier farm located near Gansbaai (33° S 18° E, 39 m.a.s.l), and at Kanetberg farm near Barrydale (33° S 21° E, 830 m.a.s.l). The 15 N natural abundance technique was used to determine N 2 fixation, carbon (C) assimilation and δ 13 C in shoot of Cyclopia and Aspalathus species.Important findings Older tea plantations of C. genistoides and C. subternata derived more N from fixation and exhibited greater water-use efficiency than younger plants. At Koksrivier, Aspalathus caledonensis and A. aspalathoides showed greater water-use efficiency and derived more N from fixation than Cyclopia genistoides. Annual harvesting of C. genistoides decreased N 2 fixation. At Kanetberg, C. subternata plants on the upper and middle slopes derived more N from atmospheric fixation than those on the lower slope. C. subternata plants grown from seedlings recorded greater %Ndfa than cuttings. N 2 fixation and water-use efficiency of Cyclopia was affected by age, slope and planting material. Further, symbiotic N nutrition and water-use efficiency of Cyclopia and Aspalathus were related.     

Factors Associated with Increased Mortality in a Predominantly HIV-Infected Population with Stevens Johnson Syndrome and Toxic Epidermal Necrolysis

Introduction

Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are life-threatening drug reactions with a higher incidence in HIV-infected persons. SJS/TEN are associated with skin and mucosal failure, predisposing to systemic bacterial infection (BSI), a major cause of death. There are limited data on risk factors associated with BSI and and mortality in HIV-infected people with SJS/TEN.

Methods

We conducted a retrospective study of patients admitted to a university hospital with SJS/TEN over a 3 year period. We evaluated their underlying illnesses, eliciting drugs, predictive value of bacterial skin cultures and other factors associated with mortality and BSI in a predominantly HIV-infected population by comparing characteristics of the patients who demised and those who survived.

Results

We admitted 86 cases during the study period and 67/86(78%) were HIV-infected. Tuberculosis was the commonest co-morbidity, diagnosed in 12/86(14%) cases. Skin cultures correlated with BSI by the same organism in 7/64(11%) cases and 6/7 were Gram-negative. Two of the 8 cases of Gram-negative BSI had an associated Gram-negative skin culture, although not always the same organism. All 8 fatalities had >30% epidermal detachment, 7 were HIV-infected, 6 died of BSI and 6 had tuberculosis.

Conclusions

Having >30% epidermal detachment in SJS/TEN carries an increased risk of BSI and mortality. Tuberculosis and BSI are associated with higher risk of death in SJS/TEN. Our data suggests there may be an association between Gram-negative BSI and Gram-negative skin infection.