First Syntheses of Melophlins P,Q, and R,and Effects of Melophlins on the Growth of Microorganisms and Tumor Cells

The marine tetramic acid (=1,5‐dihydro‐4‐hydroxy‐2H‐pyrrol‐2‐ones) derivatives melophlin P, Q, and R ( 1p – 1r , resp.) were synthesized for the first time in only four steps. Together with the congenerous melophlins A–C and G, they were also tested for antimicrobial and cytotoxic effects. Melophlins B, C, P, Q, and R, which share a 5‐Me residue, showed some antibacterial activity, mainly in Gram‐positive bacteria. Melophlins B, C, and R, which have Me‐branched 3‐acyl side chains in common, inhibited the growth of cells of human KB‐3‐1 cervix carcinoma, A‐498 kidney carcinoma, and U‐937 leukemia with IC₅₀ values <10 μM . They were similar in activity to cisplatin. Melophlin Q, also Me‐branched, was astoundingly specific in inhibiting A‐498 kidney cancer cells, while melophlin P inhibited U‐937 leukemia cells particularly well. The position of the Me branch is decisive for the magnitude of the antiproliferative effect of the melophlin couples B/C and R/Q.     

Temperature alternations and the influence of gibberellins and indoleacetic acid on elongation growth and flowering of Begonia x hiemalis Fotsch

The aim of this study was to investigate the role of plant hormones, particularly the gibberellins (GAs), in the thermoperiodic regulation of stem elongation in the short day plant (SDP) Begonia x hiemalis. Effects of GAs and some GA precursors were tested on plants grown under alternating day/night temperatures (DT/NT; 12/12 h), and the effects of these temperature regimes on endogenous plant hormones were analyzed using combined gas chromatography and mass spectrometry (GC-MS).Compared with constant temperatures (19/19 °C; 21/21 °C), stem elongation was significantly inhibited by low DT/high NT (14/24 °C; 18/24 °C) and enhanced by the opposite treatments (24/14 °C; 26/17 °C). GA1 stimulated elongation of internodes and petioles while ent-kaurene, kaurenoic acid, GA12, GA19, GA20 had no significant effect. The effect of GA1 was enhanced by a simultaneous application of calcium 3,5-dioxo-4-propionylcyclohexanecarboxylate (BX-112). BX-112 inhibited internode elongation at high DT/low NT (24/14 °C) but not at the reverse temperature regime.Gibberellins A53, A19, A20, A1, A4, A9, and indoleacetic acid (IAA), were identified by GC-MS from both leaves, including the petioles, and stems of B. x hiemalis. There were no apparent relationships between elongation of internodes and petioles and endogenous contents of gibberellins A53, A19, A20, and A1. Recoveries of deuterated GA4 and GA9 were generally too low for estimation of endogenous levels of these GAs.Constant temperature resulted in more open flowers and flower buds compared to alternating DT and NT. BX-112 decreased the time to anthesis.     

Acute regulation of Na/H exchanger NHE3 activity by protein kinase C: role of NHE3 phosphorylation

Acute hormonalmodulation of NHE3 activity is partly mediated by kinases, includingprotein kinase C (PKC). We examined the role of NHE3 phosphorylation inregulating its activity in response to PKC activation by phorbol12-myristate 13-acetate (PMA). In pooled NHE-deficient fibroblaststransfected with NHE3, PMA increased NHE3 activity and phosphorylation.When six potential PKC target serines were mutated, NHE3phosphorylation was drastically reduced and PMA failed to regulate NHE3phosphorylation or function. To examine whether NHE3 phosphorylation issufficient for functional regulation by PKC, we exploited theheterogeneous response of NHE3 activity to PMA in individual clones oftransfectants. Clones with stimulatory, inhibitory, or null responsesto PMA were observed. Despite the diverse functional response, changesin NHE3 phosphorylation as revealed by tryptic phosphopeptide maps weresimilar in all clones. We conclude that although phosphorylationappears to be necessary, it is insufficient to mediate PKC regulationof NHE3 function and factors extrinsic to the NHE3 protein must be involved.     

High-Level Production of Human Parathyroid Hormone (hPTH) by Induced Expression inSaccharomyces cerevisiae

Saccharomyces cerevisiaewas used as host for high-level production of intact human parathyroid hormone (hPTH). The yield increased about 30-fold by changing from the constitutive MFα promoter to the inducibleCUP1promoter in the expression cassettes, use of another host strain, and optimization of growth conditions where especially the pH value was crucial. The secreted products consisted mainly of intact hormone, hPTH(1-84). In addition, two C-terminally truncated forms that lacked the four or five last amino acid residues, hPTH(1-80) and hPTH(1-79), were identified. These hPTH forms migrated aberrantly by SDS–PAGE as 14-kDa proteins, while the real masses measured by mass spectrometry on HPLC-purified products were about 9 kDa. Availability of such easily purified truncated forms will be valuable for studies of how the C-terminal residues affect the structure and function of the hormone. Combination of mutations and disruptions of the host genes encoding proteinase A, B, carboxypeptidase Y, and Kex1p or Mkc7p did not influence the C-terminal deletions. The secretion of hPTH could be enhanced by overexpression of the yeast syntaxin geneSSO2, but the total level of the hormone was not improved due to impaired growth.     

Influences of fallow age and season on the foraging behavior and diet selection pattern of goats (Capra hircus L.)

A study was undertaken to determine the foraging behavior and diet selection pattern of goats maintained on communal exclosures in relation to the influence of fallow age (years since effective protection initiated; <8 years vs. >12 years) and season (long rainy, dry, and short rainy seasons). The frequency of browsing was higher (P < 0.05) in the exclosures fallowed for >12 years compared to those fallowed for <8 years and this frequency was also highest during the short rainy season. More (P < 0.05) herbaceous bites were recorded in the <8 years fallow exclosures, compared to the >12 years fallow exclosures. Herbaceous species bites were maximized during the long rainy season whereas woody species bites were maximized during the dry season. The goats foraged upon 48 plant species of which 69% were woody species, 19% grasses, and the rest comprised of various forb species. The top species that significantly (P < 0.05) contributed to the diet of goats were Acacia etbaica, Balanites aegyptica, Cadaba farinosa, Capparis tomentosa, Dichrostachys cinerea, and Ocimum forskolei. The goats targeted taller woody species than the average of the exclosures. Maurea angolensis, Jasminium abyssinicum, C. farinosa, and Ormocarpum pubescence were among the preferred species that appeared to be on the verge of disappearance in most of the exclosures investigated in this study and require immediate attention. Using goats to reduce the threat of potential bush infestation appears to be a viable option for managing and sustaining production from exclosures since the goats showed preference for D. cinerea, the species considered a prolific invader plant. There is, however, a need to conduct further research on the impacts of direct browsing by the goats and this should also entail the determination of optimum number of goats/unit area.     

Haematological complications in polycythaemia vera and thrombocythaemia patients treated with radiophosphorus (32P).

We have evaluated 230 patients with myeloproliferative disorders treated in the last 15 years with 32P. None of the patients affected by essential thrombocythaemia developed haematological complications. In the larger group of polycythaemia patients (214 subjects) only 38 patients (17 males and 21 females) developed complications. 60.5% of these subjects had a minor complications: 1.8% showed a thrombocytopenia lower than 100.10e9/lt, 2.3% anaemia with Hb lower than 10 g%, 2.6% leukopenia lower than 40.10e9/lt and 2.3% a pancytopenia. All these complications were transient and eventually treated with limited blood transfusions. We could not identify a correlation between the dose used and the development of such complications. We noted only that the occurrence of anaemia, given a similar dose, was more frequent in females. Only 7% of all patients presented a major complication after 32P administration. In this case too, there was no correlation with the dose administered. Myelofibrosis and chronic myeloid leukaemia resulted to be the more frequent complication (9 out of 15) but we could not clarify if they represented a natural evolution of polycythaemia vera or were due to the treatment with 32P. Acute leukaemia developed only in 5 patients and again we could not recognized a correlation with the dose administered. Moreover, the time from the diagnosis of polycythaemia vera the onset of acute leukaemia ranged widely. 32P has a definite effect on the prevention of thrombotic and haemorrhagic complications in polycythaemia patients since it prolongs their life but it also increases the incidence of acute leukaemia.