Mining the 99 Lives Cat Genome Sequencing Consortium database implicates genes and variants for the Ticked locus in domestic cats (Felis catus).

Tabby patterns of fur coats are defining characteristics in wild and domestic felids. Historically, three autosomal alleles at one locus (Tabby): Abyssinian (Ta ; a.k.a. ticked), mackerel (Tm ; a.k.a. striped) and blotched (tb ; a.k.a. classic, blotched) were thought to control these patterns in domestic cats and their breeds. Currently, at least three loci influence cat tabby markings, two of which are designated Tabby and Ticked. The Tabby locus is laeverin (LVRN) and affects the mackerel and blotched patterns. The unidentified gene for the Ticked locus on cat chromosome B1 was suggested to control the presence or absence of the ticked pattern (Tabby - Abyssinian (Ta ; a.k.a. ticked). The cat reference genome (Cinnamon, the Abyssinian) has the ticked phenotype and the variant dataset and coat phenotypes from the 99 Lives Cat Genome Consortium (195 cats) were used to identify candidate genes and variants associated with the Ticked locus. Two strategies were used to find the Ticked allele(s), one considered Cinnamon with the reference allele or heterozygous (Strategy A) and the other considered Cinnamon as having the variant allele or heterozygous (Strategy B). For Strategy A, two variants in Dickkopf Wnt Signaling Pathway Inhibitor 4 (DKK4), a p.Cys63Tyr (B1:41621481, c.188G>A) and a less common p.Ala18Val (B1:42620835, c.53C>T) variant are suggested as two alleles influencing the Ticked phenotype. Bioinformatic and molecular modeling analysis suggests that these changes disrupt a key disulfide bond in the Dkk4 cysteine-rich domain 1 or Dkk4 signal peptide cleavage respectively. All coding variants were excluded as Ticked alleles using Strategy B.

Standardization of a SNP panel for parentage verification and identification in the domestic cat (Felis silvestris catus).

The domestic cat (Felis silvestris catus) is a valued companion animal throughout the world. Over 60 different cat breeds are accepted for competition by the cat fancy registries in different countries. Genetic markers, including short tandem repeats and SNPs, are available to evaluate and manage levels of inbreeding and genetic diversity, population and breed structure relationships, and individual identification for forensic and registration purposes. The International Society of Animal Genetics (ISAG) hosts the Applied Genetics in Companion Animals Workshop, which supports the standardization of genetic marker panels and genotyping for the identification of cats via comparison testing. SNP panels have been in development for many species, including the domestic cat. An ISAG approved core panel of SNPs for use in cat identification and parentage analyses is presented. SNPs (n = 121) were evaluated by different university-based and commercial laboratories using 20 DNA samples as part of the ISAG comparison testing procedures. Different SNP genotyping technologies were examined, including DNA arrays, genotyping-by-sequencing and mass spectroscopy, to select a robust and efficient panel of 101 SNPs as the ISAG core panel for cats. The SNPs are distributed across all chromosomes including two on the X chromosome and an XY pseudo-autosomal sexing marker (zinc-finger XY; ZFXY). A population study demonstrated that the markers have an average polymorphic information content of 0.354 and a power of exclusion greater than 0.9999. The SNP panel should keep testing affordable while also allowing for the development of additional panels to monitor health, phenotypic traits, hybrid cats and highly inbred cats.

Validation and efficacy of transgenerational mass marking of otoliths in viviparous fish larvae.

Transgenerational mass marking of viviparous fish larvae in vivo was validated by intra-muscular injection of elemental strontium chloride (SrCl(2)) in gestating females and detection of the Sr in the otoliths of developing larvae. All otoliths of brown rockfish Sebastes auriculatus larvae produced from SrCl(2)-injected females showed enriched Sr:Ca ratios near the otolith edges, and the signatures did not appear to be affected by the anterior, centre and posterior positions of larvae within the ovary. Results from the present study indicate that transgenerational marking is a highly reliable technique for marking large numbers of extremely small viviparous fish larvae.

Assisted reproduction mediated resurrection of a feline model for Chediak-Higashi syndrome caused by a large duplication in LYST.

Chediak-Higashi Syndrome (CHS) is a well-characterized, autosomal recessively inherited lysosomal disease caused by mutations in lysosomal trafficking regulator (LYST). The feline model for CHS was originally maintained for ~20 years. However, the colonies were disbanded and the CHS cat model was lost to the research community before the causative mutation was identified. To resurrect the cat model, semen was collected and cryopreserved from a lone, fertile,  CHS carrier male. Using cryopreserved semen, laparoscopic oviductal artificial insemination was performed on three queens, two queens produced 11 viable kittens. To identify the causative mutation, a fibroblast cell line, derived from an affected cat from the original colony, was whole genome sequenced. Visual inspection of the sequence data identified a candidate causal variant as a ~20 kb tandem duplication within LYST, spanning exons 30 through to 38 (NM_001290242.1:c.8347-2422_9548 + 1749dup). PCR genotyping of the produced offspring demonstrated three individuals inherited the mutant allele from the CHS carrier male. This study demonstrated the successful use of cryopreservation and assisted reproduction to maintain and resurrect biomedical models and has defined the variant causing Chediak-Higashi syndrome in the domestic cat.

Propranolol antagonizes the anti-inflammatory effect of alcohol and improves survival of infected intoxicated rabbits.

We studied the effects of alcohol and propranolol on the course of peritonitis in rabbits. Induction of sterile peritonitis with normal saline led to a 50% augmentation of granulocyte adherence in normal rabbits, and a mean cumulative granulocyte count of 27,000/mm(3) in peritoneal exudate by 8 h. Rabbits intoxicated with alcohol at the time of peritonitis induction maintained a granulocyte adherence below pretreatment values, and only delivered a cumulative mean of 12,000 granulocytes/mm(3) into the peritoneal fluid. When intoxicated rabbits received propranolol intravenously at the time of intoxication, adherence increased above preperitonitis levels, and stayed significantly above values for animals given alcohol alone. In addition, the defect in granulocyte delivery was prevented by propranolol, resulting in a mean cumulative granulocyte count in peritoneal fluid of 24,000/mm(3).When peritonitis was induced with live pneumococci instead of a sterile inflammatory stimulus, 14/18 normal animals survived the infection and were culture-negative when sacrificed at 2 wk. In contrast, 17/18 intoxicated animals died of the infection, in a mean of 2.8 days. 9 of 18 intoxicated animals who also received propranolol survived, and those who died lived a mean of 7.5 days. The survival rates and the time-to-death among the nonsurvivors given propranolol were both significantly greater than in the animals intoxicated without propranolol. Thus, propranolol prevents the granulocyte adherence and delivery defects induced by alcohol intoxication, and significantly improves survival from infection.

Persistent meningeal inflammation associated with intrathecal gentamicin.

A 30-year-old woman was treated with intraventricular gentamicin for Klebsiella meningitis. Cultures became sterile, but meningeal inflammation, associated with high central spinal fluid gentamicin levels, persisted until gentamicin was discontinued.

Parenteral and oral fluconazole for acute cryptococcal meningitis in AIDS: experience with thirteen patients.

OBJECTIVE: Cryptococcus neoformans infections of the central nervous system affect up to ten percent of AIDS patients. Standard therapy with amphotericin B with or without 5-flucytosine has a high rate of failure, relapse, and toxicity. Fluconazole is a new triazole antifungal agent available in both oral and intravenous forms that has shown efficacy in the primary and maintenance treatment of cryptococcal meningitis in AIDS patients. In this open, noncomparative trial, we evaluated the safety and efficacy of intravenous fluconazole followed by oral fluconazole in the treatment of acute cryptococcal meningitis in AIDS patients. METHODS: Thirteen AIDS patients with acute cryptococcal meningitis, or relapse after successful primary therapy, received 400 mg of intravenous fluconazole daily for 12-16 days followed by oral fluconazole 400 mg/d for the duration of primary therapy. If cerebrospinal fluid (CSF) cultures converted to negative within 32 weeks of treatment, the fluconazole dose was decreased to 200 mg/d as maintenance therapy. RESULTS: Fluconazole therapy was successful in six patients (46 percent) and unsuccessful in seven (54 percent). Of the seven patients considered unsuccessful, one demonstrated clinical improvement but remained CSF-culture positive, five were clinical failure and were switched to amphotericin B therapy, and one died after two weeks secondary to cryptococcal meningitis. No patient experienced any adverse reactions necessitating discontinuation of therapy. CONCLUSIONS: In this small group of patients, moderate doses of parenteral and oral fluconazole for acute cryptococcal meningitis in AIDS patients demonstrated failure rates similar to those reported in other studies with fluconazole and with amphotericin B. As there was no difference in initial Karnofsky scores or the severity of disease in treatment successes versus failures, it is difficult to determine who might respond to fluconazole as initial therapy or who should be treated initially with another agent. Further studies and clinical experience are needed.

Infrainguinal anastomotic arterial graft infections treated by selective graft preservation.

The purpose of this study was to determine whether the type of graft material and bacteria involved in an infrainguinal arterial anastomotic infection can be used as guidelines for graft preservation. Between 1972 and 1990, the authors treated 35 anastomotic infections involving a common femoral or distal artery. The graft material was Dacron in 14 patients, polytetrafluoroethylene (PTFE) in 14, and vein in 7. Of the 14 Dacron grafts, immediate graft excision was required for overwhelming infection in eight patients (bleeding in five, sepsis in three) and for an occluded graft in one patient. Three of five patients failed attempted graft preservation because of nonhealing wounds. Thus, 12 of the 14 Dacron grafts ultimately required graft excision. Of the 21 "smooth-walled" vein and PTFE grafts, 10 required immediate graft excision for occluded grafts (five PTFE, one vein) or bleeding (three PTFE, one vein). Ten of the remaining 11 (91%) patients with patent "smooth-walled" grafts, intact anastomoses, and absence of sepsis managed by graft preservation healed their wounds and maintained distal arterial perfusion. Wound cultures grew pure gram-positive cocci in 17 of 21 "smooth-walled" graft infections versus 8 of 14 Dacron graft infections. In the absence of systemic sepsis, graft preservation is the treatment of choice for gram-positive infections involving an intact anastomosis of patent PTFE and vein grafts. Regardless of the bacterial cause, the authors recommend that any infrainguinal anastomotic infection of a Dacron graft be treated by immediate excision of all infected graft material.

Syphilitic uveitis in human immunodeficiency virus-infected and noninfected patients.

Seventeen patients with luetic uveitis had human immunodeficiency virus (HIV) testing and lumbar puncture to determine if the HIV-positive status altered the clinical profile of syphilis. Twelve of the 17 patients tested positive for HIV. All 12 patients had abnormal lumbar punctures, but only two of the five HIV-negative patients had abnormal results of spinal taps. Ocular disease also was more severe in the HIV-positive group.

Treatment of the septic hip with total hip arthroplasty.

Forty-three patients, 23 with definite infection and 20 with probable infections before total hip arthroplasty (THA), were compared to 41 matched uninfected patients. The 43 infected patients were treated by 45 operative procedures: eight Girdlestone resections, 12 revisions of total hips, and 25 conversions from infected nontotal hip surgery to total hip arthroplasties. (Two revision THAs were converted to Girdlestones). The average follow-up period was 38.8 months, with a range of six-118 months. The statistically significant negative prognosticators were gross sepsis at surgery, number of previous operations, and elevated erythrocyte sedimentation rate (ESR). The type of infecting organism did not affect the outcome. The prosthesis survival rate for total hip arthroplasties revised for sepsis was 83%. The prosthesis survival rate for other infected hips treated by total hip arthroplasty was 100%. All groups except Girdlestone resections improved postoperatively. While Girdlestone resection offered acceptable pain relief, total hip arthroplasty provided unequivocally superior function (p = 0.0001).