title: Bioactive Chemicals of Importance in Endophyte-Infected Grasses

authors: Babu, Jacob

subjects: indole-diterpenoids

abstract: Janthitrems are believed to be involved in the observed sporadic cases of AR37-
infected perennial ryegrass staggers. Investigations into the role of janthitrems in
perennial ryegrass staggers are difficult as isolation of the compounds from the
ryegrass is hindered by the inherent instability of these compounds. Therefore
attempts were made to isolate janthitrems from an alternative source, allowing
these janthitrem analogues to be used as surrogates for endophyte produced
janthitrems.
Analysis of a series of Penicillium janthinellum cultures revealed the presence of
janthitrems in a number of strains, including janthitrem B, janthitrem C and two
novel janthitrem compounds. Detailed one- and two-dimensional NMR and mass
spectral techniques identified the two novel compounds as 11,12-
epoxyjanthitrems B and C, which were subsequently given the trivial names
janthitrems A and D, respectively. Janthitrems B and C were isolated and
identified by NMR and revisions of some previously reported chemical shift
assignments were proposed. In addition to the janthitrems, penitrems were also
identified in two strains of P. janthinellum.
The isolated janthitrem B was utilised for the development of efficient extraction
procedures, and for the determination of ideal storage conditions for janthitrem
compounds. A method for the extraction and isolation of janthitrem B from a P.
janthinellum culture was developed and optimised to yield 6 mg of janthitrem B
from 900 mL of fungal culture in two days. Stability studies of janthitrem B
indicated the ideal storage condition which minimised degradation was dry at
−80 C where only 7% sample loss was observed over 300 days.
Bioactivity studies of janthitrems A and B found these compounds to be
tremorgenic to mice, with janthitrem A (an epoxyjanthitrem) inducing more
severe tremors than janthitrem B. Insect testing also showed that both janthitrems
A and B displayed anti-insect activity to porina larvae. Since the
epoxyjanthitrems, which are associated with AR37 endophyte-infected ryegrass,
were also shown to be tremorgenic and to display anti-insect activity, the insect
resistance and the sporadic cases of ryegrass staggers displayed by AR37 may be
related to the presence of epoxyjanthitrem compounds.
LC-UV-MS analysis of janthitrems A-D, penitrems A-F, lolitrem B, paspalinine,
paxilline and terpendole C found these indole-diterpenoids to be more sensitive
by analysis using an APCI source as opposed to an ESI source. APCI negative
ion LC-UV-MS required source induced dissociation in combination with
increased collision energy to suppress an acetate adduct peak, sourced from the
acetic acid buffer. Negative ion MS2 and MS3 data produced more informative
fragments compared to the conventional positive ion MS2 and MS3 data. The
availability of both positive and negative ion LC-UV-MS methodologies will
allow future endophyte products to be more thoroughly screened for different
classes of secondary metabolites.
Extracts of mouldy walnuts were analysed for the presence of tremorgenic
mycotoxins after a dog was found to exhibit symptoms characteristic of
tremorgenic mycotoxicosis. LC-UV-MS analysis of the mouldy walnuts
identified the tremorgenic mycotoxins penitrems A-F, thus confirming the
veterinarian's tentative diagnosis of canine tremorgenic mycotoxicosis   the first
reported case in New Zealand.

text: Bioactive Chemicals of Importance in Endophyte-Infected Grasses Janthitrems are believed to be involved in the observed sporadic cases of AR37-
infected perennial ryegrass staggers. Investigations into the role of janthitrems in
perennial ryegrass staggers are difficult as isolation of the compounds from the
ryegrass is hindered by the inherent instability of these compounds. Therefore
attempts were made to isolate janthitrems from an alternative source, allowing
these janthitrem analogues to be used as surrogates for endophyte produced
janthitrems.
Analysis of a series of Penicillium janthinellum cultures revealed the presence of
janthitrems in a number of strains, including janthitrem B, janthitrem C and two
novel janthitrem compounds. Detailed one- and two-dimensional NMR and mass
spectral techniques identified the two novel compounds as 11,12-
epoxyjanthitrems B and C, which were subsequently given the trivial names
janthitrems A and D, respectively. Janthitrems B and C were isolated and
identified by NMR and revisions of some previously reported chemical shift
assignments were proposed. In addition to the janthitrems, penitrems were also
identified in two strains of P. janthinellum.
The isolated janthitrem B was utilised for the development of efficient extraction
procedures, and for the determination of ideal storage conditions for janthitrem
compounds. A method for the extraction and isolation of janthitrem B from a P.
janthinellum culture was developed and optimised to yield 6 mg of janthitrem B
from 900 mL of fungal culture in two days. Stability studies of janthitrem B
indicated the ideal storage condition which minimised degradation was dry at
−80 C where only 7% sample loss was observed over 300 days.
Bioactivity studies of janthitrems A and B found these compounds to be
tremorgenic to mice, with janthitrem A (an epoxyjanthitrem) inducing more
severe tremors than janthitrem B. Insect testing also showed that both janthitrems
A and B displayed anti-insect activity to porina larvae. Since the
epoxyjanthitrems, which are associated with AR37 endophyte-infected ryegrass,
were also shown to be tremorgenic and to display anti-insect activity, the insect
resistance and the sporadic cases of ryegrass staggers displayed by AR37 may be
related to the presence of epoxyjanthitrem compounds.
LC-UV-MS analysis of janthitrems A-D, penitrems A-F, lolitrem B, paspalinine,
paxilline and terpendole C found these indole-diterpenoids to be more sensitive
by analysis using an APCI source as opposed to an ESI source. APCI negative
ion LC-UV-MS required source induced dissociation in combination with
increased collision energy to suppress an acetate adduct peak, sourced from the
acetic acid buffer. Negative ion MS2 and MS3 data produced more informative
fragments compared to the conventional positive ion MS2 and MS3 data. The
availability of both positive and negative ion LC-UV-MS methodologies will
allow future endophyte products to be more thoroughly screened for different
classes of secondary metabolites.
Extracts of mouldy walnuts were analysed for the presence of tremorgenic
mycotoxins after a dog was found to exhibit symptoms characteristic of
tremorgenic mycotoxicosis. LC-UV-MS analysis of the mouldy walnuts
identified the tremorgenic mycotoxins penitrems A-F, thus confirming the
veterinarian's tentative diagnosis of canine tremorgenic mycotoxicosis   the first
reported case in New Zealand.

year: 2009

title: Methods of a national colorectal cancer cohort study: the PIPER Project

authors: Firth, Melissa

abstract: A national study looking at bowel cancer in New Zealand has previously been completed (the PIPER Project). The study included 5,610 patients and collected medical information about how each person was found to have bowel cancer and the treatment they received. This paper reports how the study was carried out. The information collected in the study will be used to look at the quality of care being provided to New Zealand patients with bowel cancer, and to find out if differences in care occur based on where people live, their ethnicity and their socioeconomic status.

text: Methods of a national colorectal cancer cohort study: the PIPER Project A national study looking at bowel cancer in New Zealand has previously been completed (the PIPER Project). The study included 5,610 patients and collected medical information about how each person was found to have bowel cancer and the treatment they received. This paper reports how the study was carried out. The information collected in the study will be used to look at the quality of care being provided to New Zealand patients with bowel cancer, and to find out if differences in care occur based on where people live, their ethnicity and their socioeconomic status.

year: 2016

title: Developing a kaupapa Māori evaluation model – one size fits all?

authors: Masters-Awatere, Bridgette

subjects: Maori health

abstract: Health statistics in Aotearoa (New Zealand) highlight that Māori, the indigenous people Aotearoa have poorer health than non- Māori. In response to the statistics a number of Māori health providers have established services that address specific areas of need in their regions. Initially there were minimal accountability requirements of providers. However, changes in the health system now mean that groups wanting to establish a new service must provide accountability measures before, during and after the funding has been allocated. As a result providers need to develop a rationale behind their decisions and assess the measure of change that has taken place as a result of the service or programme to ensure continued funding. The requirements reflect the dominant Western paradigm in which health promotion is understood to be about producing specific quantifiable behaviour changes in individuals. Māori health providers on the other hand have tended to take a holistic approach to health. Thus they have found themselves in the position of trying to show change within a paradigm where measurements are not easily taken. This has created frustration amongst Māori providers who face losing their funding because of an inability to report measurable outcomes using a framework that does not apply to their culture.

text: Developing a kaupapa Māori evaluation model – one size fits all? Health statistics in Aotearoa (New Zealand) highlight that Māori, the indigenous people Aotearoa have poorer health than non- Māori. In response to the statistics a number of Māori health providers have established services that address specific areas of need in their regions. Initially there were minimal accountability requirements of providers. However, changes in the health system now mean that groups wanting to establish a new service must provide accountability measures before, during and after the funding has been allocated. As a result providers need to develop a rationale behind their decisions and assess the measure of change that has taken place as a result of the service or programme to ensure continued funding. The requirements reflect the dominant Western paradigm in which health promotion is understood to be about producing specific quantifiable behaviour changes in individuals. Māori health providers on the other hand have tended to take a holistic approach to health. Thus they have found themselves in the position of trying to show change within a paradigm where measurements are not easily taken. This has created frustration amongst Māori providers who face losing their funding because of an inability to report measurable outcomes using a framework that does not apply to their culture.

year: 2002