ABSTRACT
Weanling pigs tend to avoid wind, and suckling piglets are thought to be more sensitive to wind than weanlings, owing to their thinner s.c. fat layer. We developed a crush-reducing device based on the anticipated behavior of suckling piglets toward wind and evaluated the performance of the device through field testing. The crush-reducing device consisted of six photo sensors, a controller, six solenoid valves, and an air compressor. In this study, 206 sows and their suckling piglets (Landrace xYorkshire) were investigated to ascertain the individual effects of several factors. Some of the newborn piglets were cross-fostered before the crush-reducing device was implemented. Litter weights were measured on d 0 and 4 to determine the influence of the crush-reducing device on the weights of suckling piglets. The crushing of suckling piglets by sows was affected by season (P < 0.01) and litter size (P < 0.05), but not by the parity of the sows; however, the number of crushed piglets per litter was less (P < 0.01) in the crush-reducing device group (0.05 +/- 0.02 crushed piglets/litter) than in the control group (0.23 +/- 0.04 crushed piglets/litter), regardless of litter size or season. The BW gain of suckling piglets did not differ between the control and the crush-reducing device groups. Based on these results, the crush-reducing device is expected to decrease the number of crushed piglets per litter without influencing the BW gain of suckling piglets, thereby greatly contributing to the productivity of pig breeders.
Subject(s)
Animal Husbandry/instrumentation , Animal Husbandry/methods , Animals, Suckling , Swine , Animals , Female , Mortality , Seasons , WindABSTRACT
ParE is the ATP-binding subunit of topoisomerase IV (Topo IV). During topoisomerization, the ATP-binding and hydrolysis cycle must be coordinated with the cycle of DNA cleavage and religation. We have isolated three dominant-negative mutant alleles of parE that encode ParE proteins that fail to hydrolyze ATP when reconstituted with ParC to form Topo IV. ParE G110S Topo IV and ParE S123L Topo IV failed to bind ATP at all, whereas ParE T201A could bind ATP. All three mutant Topo IV proteins exhibited an elevated level of spontaneous DNA cleavage that could be associated with a decreased rate of DNA resealing. In ParE T201A Topo IV, this defect appeared to result from an increased likelihood that the tetrameric enzyme would fall apart after DNA cleavage. Thus, while ATP is not required for DNA cleavage, the properties of these mutant enzymes suggests that ATP-hydrolysis informs DNA religation.
Subject(s)
Adenosine Triphosphatases/genetics , Bacterial Proteins/genetics , DNA Topoisomerases, Type II/genetics , DNA-Binding Proteins/genetics , Escherichia coli/enzymology , Adenosine Triphosphate/metabolism , Adenylyl Imidodiphosphate/metabolism , Animals , Chromatography, Gel , Crithidia fasciculata , DNA Mutational Analysis , DNA Topoisomerase IV , DNA, Superhelical/metabolism , Dimerization , Models, Molecular , MutationABSTRACT
In order to define regions of ParE, one of the two subunits of topoisomerase IV, that are involved in catalysis during topoisomerization, we developed a selection procedure to isolate dominant-negative parE alleles. Both wild-type parC and mutagenized parE were expressed from a tightly-regulated lac promoter on a moderate-copy plasmid. Mutated parE alleles were rescued from those plasmids that caused IPTG-dependent cell death. The mutant ParE proteins could be divided into two groups when reconstituted with ParC to form topoisomerase IV, those that elicited hyper-DNA cleavage and those that affected covalent complex formation.
Subject(s)
Bacterial Proteins/genetics , DNA Topoisomerases, Type II/genetics , DNA-Binding Proteins/genetics , Escherichia coli/enzymology , Alleles , DNA Topoisomerase IV , DNA, Superhelical/chemistry , Electrophoresis, Polyacrylamide Gel , Isopropyl Thiogalactoside/metabolism , Microscopy, Fluorescence , Mutation , PhenotypeABSTRACT
The products of three dominant-negative alleles of parE, encoding the ATP-binding subunit of topoisomerase IV (Topo IV), were purified and their activities characterized when reconstituted with ParC to form Topo IV. The ability of the ParE E418K, ParE G419D, and ParE G442D mutant Topo IVs to bind DNA, hydrolyze ATP, and close their ATP-dependent clamp was relatively unaffected. However, their ability to relax negatively supercoiled DNA was compromised significantly. This could be attributed to severe defects in covalent complex formation between ParC and DNA. Thus, these residues, which are far from the active site Tyr of ParC, contribute to covalent catalysis. This indicates that a dramatic conformational rearrangement of the protein likely occurs subsequent to the binding of the G segment at the DNA gate and prior to its opening.
Subject(s)
Bacterial Proteins/genetics , DNA Topoisomerases, Type II/genetics , DNA-Binding Proteins/genetics , Escherichia coli/enzymology , Adenosine Triphosphate/metabolism , Alleles , Bacterial Proteins/chemistry , Binding Sites , DNA Topoisomerase IV , DNA Topoisomerases, Type II/chemistry , DNA, Superhelical/metabolism , DNA-Binding Proteins/chemistry , Models, Molecular , Mutation , Norfloxacin/chemistry , Protein Binding/genetics , Protein ConformationABSTRACT
OBJECTIVE: There are little data on the actual care given pediatric tracheostomy patients in their homes. Information on the use of supplies and on techniques and frequency of care is valuable for a better understanding of the needs of this population. DESIGN: Questionnaires were distributed by mail or at clinic visits from May 1995 to June 1996 to a convenience sample of tracheotomized patients at the University of Michigan Pediatric Physical Medicine and Rehabilitation clinic. SETTING: Tertiary care clinic. RESULTS: Clean technique for suctioning was reported by 96.7% of subjects and the rest reported sterile technique. Fifty percent of subjects reported reusing suction catheters. Cleaning solutions used to clean suction catheters for reuse varied. Tracheostomy tube reuse was reported by 55% of subjects. Sixty percent of those who reused tracheostomy tubes had had pneumonia within the previous year, whereas only 25% of those who never reused the tracheostomy tube had pneumonia in the same time period. CONCLUSIONS: Suctioning frequency, suction catheter, and tracheostomy tube reuse and cleaning methods are variables that warrant further investigation of safety and efficacy.
Subject(s)
Home Nursing/methods , Tracheostomy , Adolescent , Adult , Child , Child, Preschool , Disinfection , Equipment Reuse , Humans , Infant , Michigan , Suction , Surveys and QuestionnairesABSTRACT
Three phthalocyanine dyes HOSiPcOSi(CH3)2(CH2)3N(CH3)2 (Pc 4), HOSiPcOSi(CH3)2(CH2)3N+(CH3)3I- (Pc 5) and aluminum tetrasulfophthalocyanine hydroxide (AlOHPcS4) were evaluated for their ability to inactivate the trypomastigote form of Trypanosoma cruzi in fresh frozen plasma (FFP) and red blood cell concentrates (RBCC). The compound Pc 4 was found to be highly effective in killing T. cruzi, Pc 5 less effective and AlOHPcS4 ineffective. With FFP as the medium, a complete loss of parasite infectivity in vitro (> or = 5 log10) was found to occur with 2 microM Pc 4 after irradiation with red light (> 600 nm) at a fluence of 7.5 J/cm2, while with RBCC as the medium, a complete loss was found to occur at a fluence of 15 J/cm2. Even without illumination, Pc 4 at 2 microM also killed about 3.7-4.1 log10 of T. cruzi in FFP during 30 min. Observed differences in T. cruzi killing by the various phthalocyanines may related to differences in binding; Pc 4 binds to the parasites about twice as much as Pc 5. Ultrastructural analysis of treated parasites suggests that mitochondria are a primary target of this photodynamic treatment. The data indicate that Pc 4 combined with exposure to red light could be used to eliminate bloodborne T. cruzi parasites from blood components intended for transfusion. The inactivation of T. cruzi by Pc 4 in the dark suggests a possible therapeutic application.
