Biomedical subjects

Patricia Martin

Publications and source records attributed to Patricia Martin.

14 recordsLinked to original sources

PubMed2006

Development and clinical application of a panfungal PCR assay to detect and identify fungal DNA in tissue specimens.

Given the rise in the incidence of invasive fungal infections (IFIs) and the expanding spectrum of fungal pathogens, early and accurate identification of the causative pathogen is essential. We developed a panfungal PCR assay that targets the internal transcribed spacer 1 (ITS1) region of the ribosomal DNA gene cluster to detect fungal DNA in fresh and formalin-fixed, paraffin-embedded (PE) tissue specimens from patients with culture-proven (n=38) or solely histologically proven (n=24) IFIs. PCR products were sequenced and compared with sequences in the GenBank database to identify the causal pathogen. The molecular identification was correlated with results from histological examination and culture. The assay successfully detected and identified the fungal pathogen in 93.6% and 64.3% of culture-proven and solely histologically proven cases of IFI, respectively. A diverse range of fungal genera were identified, including species of Candida, Cryptococcus, Trichosporon, Aspergillus, Fusarium, Scedosporium, Exophiala, Exserohilum, Apophysomyces, Actinomucor, and Rhizopus. For five specimens, molecular analysis identified a pathogen closely related to that identified by culture. All PCR-negative specimens (n=10) were PE tissues in which fungal hyphae were visualized. The results support the use of the panfungal PCR assay in combination with conventional laboratory tests for accurate identification of fungi in tissue specimens.

DNA, Fungal↗

PubMed2006

Successful treatment of a localised CDC Group EF-4a infection in a cat.

Organisms classified within the Center for Disease Control (CDC) Group eugonic fermenter (EF)-4a are facultative anaerobic, Gram-negative coccobacilli, thought to be of the family Neisseriaceae. CDC Group EF-4a is considered a component of normal oral flora in cats and dogs and is most commonly isolated from bite injuries of human patients. Most previously reported cases in cats have involved fatal necrotising pneumonia. We report a localised infection of the retropharyngeal/mandibular tissues in a cat, which responded to surgical drainage and a 5-week course of amoxycillin clavulanate. There are no prior reports of successfully treated EF-4a infections in cats in the literature.

Animals↗

PubMed2006

Distinct pattern of lung gene expression in the Cftr-KO mice developing spontaneous lung disease compared with their littermate controls.

Cystic fibrosis (CF) is caused by a defect in the CF transmembrane conductance regulator (CFTR) protein that functions as a chloride channel. Dysfunction of the CFTR protein results in salty sweat, pancreatic insufficiency, intestinal obstruction, male infertility, and severe pulmonary disease. Most of the morbidity and mortality of CF patients results from pulmonary complications. Differences in susceptibility to bacterial infection and variable degree of CF lung disease among CF patients remain unexplained. Many phenotypic expressions of the disease do not directly correlate with the type of mutation in the Cftr gene. Using a unique CF mouse model that mimics aspects of human CF lung disease, we analyzed the differential gene expression pattern between the normal lungs of wild-type mice (WT) and the affected lungs of CFTR knockout mice (KO). Using microarray analysis followed by quantitation of candidate gene mRNA and protein expression, we identified many interesting genes involved in the development of CF lung disease in mice. These findings point to distinct mechanisms of gene expression regulation between mice with CF and control mice.

Animals↗

PubMed2006

Pilus-mediated adhesion of Neisseria meningitidis is negatively controlled by the pilus-retraction machinery.

The type IV pili (Tfp) of Neisseria meningitidis play an essential role in meningococcal virulence by mediating the initial interaction of bacteria with host cells. Tfp are also subject to retraction, which relies on the PilT protein. Among the other components of the Tfp machinery, PilC1, a pilus-associated protein, is important for Tfp biogenesis and adhesion. Adhesion of N. meningitidis to living epithelial cells was previously shown to rely on the upregulation of the pilC1 gene. On the other hand the lack of induction of pilC1 is believed to be responsible for the low adhesion of N. meningitidis onto fixed dead cells. Surprisingly, a pilT mutant, unable to retract its pili, has been shown to adhere very efficiently onto both living and fixed epithelial cells. To elucidate the mechanisms by which the pilus retraction machinery mediates meningococcal adhesion onto fixed cells, an analysis of gene expression levels in wild-type and pilT meningococci was performed using DNA microarrays. One of the upregulated genes in the pilT strain was pilC1. This result was confirmed using quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) and immunoblot analysis. The transcription starting point responsible for the upregulation of pilC1 in a pilT background was shown to be different from those controlling the induction of pilC1 upon contact with living host cells. Subsequent work using a strain hyperproducing PilT confirmed that PilT downregulates the production of PilC1. Furthermore using a pilC1 allele under the control of IPTG, we demonstrated that the upregulation of pilC1 in a pilT background was responsible for the adhesive phenotype onto fixed dead cells. Taken together our results demonstrate that the pilus retraction machinery negatively controlled the adhesiveness of the Tfp via the expression of pilC1.

Adenosine Triphosphatases↗

PubMed2005

Feline cholecystitis and acute neutrophilic cholangitis: clinical findings, bacterial isolates and response to treatment in six cases.

Clinicopathological findings from six cats with confirmed cholecystitis or acute neutrophilic cholangitis are presented. Historical findings included lethargy and anorexia or inappetence of up to five days duration. On physical examination all cats were pyrexic and four out of six were jaundiced and had cranial abdominal pain. Bile samples were obtained by cholecystocentesis at exploratory coeliotomy (two cases) or by percutaneous, ultrasound-guided cholecystocentesis (four cases). Gall bladder rupture and bile peritonitis occurred subsequent to ultrasound-guided cholecystocentesis in one case. The most common bacterial isolate was Escherichia coli (four cases); E coli was isolated alone in two cases, in combination with a Streptococcus species (one case) and in combination with a Clostridium species (one case). Streptococcus species alone was isolated from one case, as was Salmonella enterica serovar Typhimurium. The latter is the first reported case of Salmonella-associated cholecystitis in a cat. Concurrent pancreatic or intestinal disease was detected histologically in three cases. All cases were treated with antimicrobials based on in vitro susceptibility results. Treatment was successful in five cases. One cat with concurrent diffuse epitheliotropic intestinal lymphoma was euthanased. Percutaneous ultrasound-guided cholecystocentesis is an effective, minimally-invasive technique enabling identification of bacterial isolates in cats with inflammatory hepatobiliary disease.

Acute Disease↗

PubMed2005

Disseminated Mycobacterium avium infection in young cats: overrepresentation of Abyssinian cats.

Disseminated Mycobacterium avium-intracellulare complex (MAC) infection was diagnosed in 10 young cats (1-5 years of age) from Australia or North America between 1995 and 2004. A further two cats with disseminated mycobacteriosis (precise agent not identified) were recognised during this period. Of the 12, 10 were Abyssinian cats, one was a Somali cat and one was a domestic shorthair cat. None of the cats tested positive for either FeLV antigen or FIV antibody. The clinical course of these infections was indolent, with cats typically presenting for weight loss, initially in the face of polyphagia, with a chronicity of up to several months. Additional clinical features included lower respiratory tract signs and peripheral lymphadenomegaly. A marked diffuse interstitial pattern was evident in thoracic radiographs, even in cats without overt respiratory involvement. Hair clipped to perform diagnostic procedures tended to regrow slowly, if at all. Diagnosis was generally made by obtaining representative tissue specimens from mesenteric lymph nodes, liver or kidney at laparotomy, or from a popliteal lymph node. The primary antecedent event was most likely colonisation of either the alimentary or respiratory tract, followed by local invasion and eventual lymphatic and haematogenous dissemination. Nine cases were treated using combination therapy with agents effective for MAC infection in human patients. Two cats are still undergoing initial therapy and have responded. Of the remaining seven, all responded during long courses (5-14 months) of clarithromycin combined with either clofazimine or rifampicin, and a fluoroquinolone or doxycycline. Of these, three cats remain well (with durations between 2 months and 2 years following therapy); two developed recurrent disease (at 3 months and 2 years, respectively, following therapy) and have restarted therapy. The remaining two cats improved 1 year and 5 months, respectively, after diagnosis but ultimately succumbed. The two cats in which therapy was restarted have improved dramatically. Certain lines of Abyssinian and Somali cats likely suffer from a familial immunodeficiency that predisposes them to infection with slow-growing mycobacteria such as MAC.

Animals↗

PubMed2005

Feline pyothorax: a retrospective study of 27 cases in Australia.

Pyothorax was diagnosed in 27 cats between 1983 and 2002. In 21 (78%) of the cases, pleural fluid culture and/or cytology was consistent with a mixed anaerobic bacterial infection of oropharyngeal origin. In six cases (22%), infection was caused by unusual pathogens or pathogens of non-oropharyngeal origin, including a Mycoplasma species, Cryptococcus gattii, Escherichia coli, Salmonella typhimurium and Staphylococcus aureus. The overall mortality rate was 22%. Treatment was successful in 18 of 19 cases (95%) where closed thoracostomy tubes were inserted. One case resolved only after thoracotomy. Actinomyces species were isolated in three cases and in contrast to dogs where thoracotomy is recommended, they were resolved with tube thoracostomy. Mechanical complications occurred in 58% of the cats with indwelling chest tubes. Probable mechanisms of pleural space infection were identified in 18 cats (67%) including haematogenous infection (n=1), direct inoculation of bacteria into the pleural space (n=1), intrathoracic oesophageal rupture (n=1) and parapneumonic extension of infection (n=15; 56%). Of the latter, perioperative aspiration was suspected in two cats, parasitic migration in two and antecedent upper respiratory tract infection was implicated in seven. Parapneumonic spread of infection after colonisation and invasion of lung tissue by oropharyngeal flora appears to be the most frequent cause of feline anaerobic polymicrobial pyothorax and contests the widespread belief that direct inoculation of pleural cavity by bite wounds is more common.

Animals↗

PubMed2005

Microsatellite instability regulates transcription factor binding and gene expression.

Microsatellites are tandemly repeated simple sequence DNA motifs widely prevalent in eukaryotic and prokaryotic genomes. In pathogenic bacteria, instability of these hypermutable loci through slipped-strand mispairing mediates the high-frequency reversible switching of phenotype expression, i.e., phase variation. Phase-variable expression of NadA, an outer membrane protein and adhesin of the pathogen Neisseria meningitidis, is mediated by changes in the number of TAAA repeats located upstream of the core promoter of nadA. Here we report that loss or gain of TAAA repeats affects the binding of the transcriptional regulatory protein IHF to the nadA promoter. Thus, phase-variable transcription of nadA potentially incorporates interplay between stochastic (mutational) and prescriptive (classical) mechanisms of gene regulation.

Amino Acid Sequence↗

PubMed2004

Infections and some other conditions affecting the skin and subcutis of the naso-ocular region of cats--clinical experience 1987-2003.

Infections of the skin or subcutis of the naso-ocular region develop through two mechanisms. Cases with lesions but without concomitant signs of nasal disease probably result from cat scratch injuries. Under certain circumstances, such lacerations result in the introduction of saprophytic microorganisms in such large numbers that host defence mechanisms are overwhelmed. This results in localised, variably invasive, disease in an otherwise immunocompetent host. An unpredictable range of organisms can give rise to such infections including a variety of fungal and bacterial genera. Causal organisms will likely vary from one geography to another as a result of differences in soil type and related environmental factors. Accordingly, procurement of appropriate tissue specimens for culture and susceptibility testing is essential to guide therapy, as these cases require medical and sometimes surgical intervention in order to effect a favourable outcome. In contrast, patients with naso-ocular lesions and concurrent signs of nasal disease have a different pathogenesis. Primary infection of the sinonasal region likely results from the inhalation of infectious propagules, with the infection subsequently penetrating overlying bones to invade the subcutaneous space. These lesions are typically the result of cryptococcosis or aspergillosis and must be distinguished from invasive nasal malignancies. An approach to the investigation and treatment of these patients is presented together with photographs of representative cases.

Animals↗

PubMed2004

Percutaneous penetration enhancers in cigarette mainstream smoke.

Percutaneous penetration enhancers (PPEs) are chemicals used to enhance the transdermal delivery of drugs. Fifty-eight of the approximately 150 PPEs used for the transdermal delivery of drugs have been reported in cigarette mainstream smoke (MS). MS is a complex aerosol of minute liquid droplets (termed the particulate phase) suspended within a mixture of gases (CO(2), CO, NO(x), etc.) and semi-volatile compounds. The gases and many of the semi-volatiles are termed the vapor phase. Twenty-nine of the 58 PPEs have been identified in MS vapor phase, 15 in the particulate phase and 14 in both the vapor and particulate phases. There is a tendency for MS PPEs to be hydrophobic, with 40 of the 58 compounds (69%) being either hydrophobic or strongly hydrophobic, and only 24% being hydrophilic. Many of the 4800 known constituents of MS are hydrophilic and would not be expected to readily cross cell membranes or penetrate tissue when delivered as single compounds. The in vivo effect on biological activity of the juxtaposition within the cigarette smoke aerosol of the large number of hydrophilic constituents with the 58 PPEs is currently unknown. As an initial step in understanding this potential complex interaction, the 58 PPEs in MS have been identified and a number of molecular parameters related to the ability to penetrate tissue have been calculated, including MS concentration, measured and calculated base ten logarithm of the octanol-water partition coefficient (Mlog P and Clog P), molecular volume (MgVol) and calculated molar refractivity (CMR).

Administration, Cutaneous↗

PubMed2004

Initial head computed tomographic scan characteristics have a linear relationship with initial intracranial pressure after trauma.

BACKGROUND: Despite current recommendations by the Brain Trauma Foundation regarding the placement of intracranial pressure (ICP) monitoring devices, advances in computed tomographic (CT) scan technology have led to the suggestion that increased ICP may be predicted by findings on admission head CT scan and that patients without such findings do not require such monitoring. A linear relationship exists between characteristics of admission head CT scan and initial ICP level, allowing for selective placement of ICP monitoring devices. METHODS: From 1997 to 2001, a retrospective review of patients admitted with a Glasgow Coma Scale (GCS) score < 8 and head CT scan who underwent ventriculostomy placement at our institution, was conducted. Patients undergoing craniotomy with evacuation of mass lesions before ventriculostomy placement were excluded. Age, sex, mechanism of injury, anoxia, osmotic treatment, presence of drugs/alcohol, initial mean arterial pressure, initial GCS score, and initial ICP were recorded. Initial head CT scans were reviewed independently by two neuroradiologists who were blinded to ICP measurements, neurosurgical treatment, patient outcome, and each other's interpretation. Initial CT scans were evaluated and scored on a 1 (normal) to 3 (abnormal) scale with respect to ventricle size, basilar cistern size, sulci size, degree of transfalcine herniation, and gray/white matter differentiation. Initial ICP readings and CT scan findings were compared to determine whether a significant linear relationship existed between the above CT scan findings and ICPs. Logistic and univariate linear regression were used to compare averaged radiologist score versus dichotomized ICP at baseline. RESULTS: Initial head CT scan characteristics show a linear relationship to baseline ICPs. These findings are associative, but are not uniformly predictive. CONCLUSION: Therefore, the current Brain Trauma Foundation recommendation of ICP monitoring in those patients presenting with a GCS score < 8 with an abnormal CT scan or a normal CT scan with age > 40 years, systolic blood pressure < 90 mm Hg, or exhibiting posturing should be followed.

Adolescent↗

PubMed2004

Involvement of genes of genome maintenance in the regulation of phase variation frequencies in Neisseria meningitidis.

In Neisseria meningitidis, the reversible expression of surface antigens, i.e. phase variation, results from changes within repeated simple sequence motifs located in coding or promoter regions of the genes involved in their biosynthesis. The mutation rates of these simple sequences, which have a major influence on the generation of phenotypic diversity, can affect the fitness of the population. The aim of the present study was to investigate the involvement of genetic factors involved (mutS and dam) and not yet analysed (drg and dinB) in the regulation of phase variation frequencies of genes associated with a variety of repeat tracts. The frequency of frameshifts occurring in the polycytidine (polyC) tracts associated with siaD, spr and lgtG and in the tetranucleotide (TAAA) repeat tract associated with nadA was determined by colony immunoblotting or using the lacZ gene as a reporter. Inactivation of mutS increased the frequency of phase variation of genes presenting homopolymeric tracts of diverse length. Overexpression of dinB enhanced the instability of the homopolymeric tract associated with siaD. Investigation of the dam locus in a population of genetically distinct N. meningitidis strains revealed that 27 % of strains associated with invasive disease contained the dam gene. In all strains where a Dam function was absent, the drg gene had been inserted into the dam locus. Disruption of dam and drg in strains representative of each genotype, i.e. dam(+)/drg and dam/drg(+), did not modify phase variation frequencies. In contrast to the effects of certain genes on homopolymeric tracts, none of the genetic factors investigated affected the stability of tetranucleotide repeat tracts.

Adenosine Triphosphatases↗

PubMed2003

Inherited mannose-binding lectin deficiency as evidenced by genetic and immunologic analyses: association with severe recurrent infections.

BACKGROUND: Mannose-binding lectin (MBL), an acute-phase serum protein of hepatic origin, plays an essential protective role in host innate immunity in targeting microbial pathogens for destruction via opsonization, enhancement of phagocytosis and complement activation. MBL deficiency, characterized by low serum MBL, has been attributed to genetic mutations in both structural and promoter regions of the gene coding for the protein. Concomitant MBL deficiency in patients with chronic immunologic disease has been associated with increased susceptibility to complicating infections that may hasten disease progression. OBJECTIVE: Few cases of inherited MBL deficiency in adults and possible associations with recurrent infection have been reported. To address this issue, we investigated the MBL profile of four generations within a single adult family whose members have experienced a variety of persistent infections. METHODS: MBL serum levels and MBL genotypes of each participating family member were ascertained by enzyme-linked immunoadsorbent assay and reverse transcriptase-polymerase chain reaction, respectively. MBL complement activation, as measured by C4b deposition against mannan-coated wells, was assayed using an enzyme-linked immunoadsorbent assay. Routine immunologic and cellular tests were carried out to evaluate the immunologic status of each family member. RESULTS: Six of the 7 family members screened carried one or more of the variant MBL alleles in their genotype and had correspondingly low serum MBL and reduced ability to affect C4b opsonization. Medical histories of the participating family members revealed an array of mild to severe recurrent infections despite no apparent immunodeficiency. CONCLUSIONS: Our studies show that MBL deficiency is an inherited characteristic and may be a crucial factor in maintaining immunologic health.

Adult↗

PubMed2002

The potential use of saliva to detect recurrence of disease in women with breast carcinoma.

BACKGROUND: Approximately 1 woman in every 10 will develop breast cancer in her lifetime. It has been shown that screening for breast cancer can reduce breast cancer mortality. The use of a saliva-based test could prove to be very useful in post-operative and/or adjunctive therapy management of breast cancer patients. METHODS: The following study was undertaken to establish the possible usefulness of the salivary protein product of the oncogene c-erbB-2 in following patients diagnosed with carcinoma of the breast. Included in this study were 25 patients with a mean age of 54 years with varying histological diagnoses and stages of carcinoma of the breast. ELISA assays for c-erbB-2 and CA 15-3 were performed on serum and stimulated whole saliva samples collected on all patients prior to any adjunct therapy or surgery and sequentially during therapy. RESULTS: The results of the GLM analyses using marker concentration as the dependent variable and treatment regimen and the serial assessments as independent variables yielded a significant overall model for both the serum (P < 0.007) and salivary (P < 0.017) c-erbB-2 markers. The model for serum c-erbB-2, however, exhibited a significant difference for treatment regimen (P < 0.001) with the chemotherapy and radiation treatment regimen being significantly different (P < 0.001) from the other treatment therapies. Time (serial assessments) was not significant. The model for the salivary c-erbB-2 marker was reversed. Treatment regimen was not significant for this model; however, time (serial assessments) was significant (P < 0.002). The serum and salivary CA 15-3 marker models yielded no significant results. Paired t-test analyses indicated that only the salivary c-erbB-2 concentrations exhibited a significant difference between the pre- and post-therapy values (t = 4.245, P < 0.0001). Additionally, salivary c-erbB-2 displayed greater percent reductions across all therapies as compared to the other markers. CONCLUSIONS: This preliminary study appears to indicate that c-erbB-2 protein expression in saliva may be a very useful diagnostic tool for measuring patient response to chemotherapy and/or surgical treatment of their disease.

Adult↗