Genetic manipulation of cardiac Hsp72 levels does not alter substrate metabolism but reveals insights into high-fat feeding-induced cardiac insulin resistance.

Heat shock protein 72 (Hsp72) protects cells against a variety of stressors, and multiple studies have suggested that Hsp72 plays a cardioprotective role. As skeletal muscle Hsp72 overexpression can protect against high-fat diet (HFD)-induced insulin resistance, alterations in substrate metabolism may be a mechanism by which Hsp72 is cardioprotective. We investigated the impact of transgenically overexpressing (Hsp72 Tg) or deleting Hsp72 (Hsp72 KO) on various aspects of cardiac metabolism. Mice were fed a normal chow (NC) or HFD for 12 weeks from 8 weeks of age to examine the impact of diet-induced obesity on metabolic parameters in the heart. The HFD resulted in an increase in cardiac fatty acid oxidation and a decrease in cardiac glucose oxidation and insulin-stimulated cardiac glucose clearance; however, there was no difference in Hsp72 Tg or Hsp72 KO mice in these rates compared with their respective wild-type control mice. Although HFD-induced cardiac insulin resistance was not rescued in the Hsp72 Tg mice, it was preserved in the skeletal muscle, suggesting tissue-specific effects of Hsp72 overexpression on substrate metabolism. Comparison of two different strains of mice (BALB/c vs. C57BL/6J) also identified strain-specific differences in regard to HFD-induced cardiac lipid accumulation and insulin resistance. These strain differences suggest that cardiac lipid accumulation can be dissociated from cardiac insulin resistance. Our study finds that genetic manipulation of Hsp72 does not lead to alterations in metabolic processes in cardiac tissue under resting conditions, but identifies mouse strain-specific differences in cardiac lipid accumulation and insulin-stimulated glucose clearance.

Analysis of the neuromuscular activity during rising from a chair in water and on dry land.

PURPOSE: The purpose of the present study was to analyze the neuromuscular responses during the performance of a sit to stand [STS] task in water and on dry land. SCOPE: 10 healthy subjects, five males and five females were recruited for study. Surface electromyography sEMG was used for lower limb and trunk muscles maximal voluntarty contraction [MVC] and during the STS task. RESULTS: Muscle activity was significantly higher on dry land than in water normalized signals by MVC from the quadriceps-vastus medialis [17.3%], the quadriceps - rectus femoris [5.3%], the long head of the biceps femoris [5.5%], the tibialis anterior [13.9%], the gastrocnemius medialis [3.4%], the soleus [6.2%]. However, the muscle activity was higher in water for the rectus abdominis [-26.6%] and the erector spinae [-22.6%]. CONCLUSIONS: This study for the first time describes the neuromuscular responses in healthy subjects during the performance of the STS task in water. The differences in lower limb and trunk muscle activity should be considered when using the STS movement in aquatic rehabilitation.

Robotic-assisted hysterectomy in a patient with a ventriculoperitoneal shunt.

There are several articles in the literature reporting laparoscopic surgery in patients with ventriculoperitoneal shunts (VPSs). Although the majority of these conclude that a pneumoperitoneum in these patients is safe, there are other reports indicating possible complications of the insufflation. This is the first known report of a robotic-assisted hysterectomy performed on a patient with a VPS and the management of the shunt during the procedure.

Ruptured tubal pregnancy associated with intrauterine pregnancy complicated by fetal holoprosencephaly: a case report.

BACKGROUND: Naturally occurring heterotopic pregnancy is rare. A surviving intrauterine pregnancy associated with a ruptured tubal pregnancy is extremely unusual. CASE: This is the first reported case of a patient presenting in hemorrhagic shock due to a ruptured tubal pregnancy that was associated with an ongoing intrauterine pregnancy complicated by fetal holoprosencephaly. CONCLUSION: Delays in diagnosis and treatment of heterotopic pregnancies may adversely affect maternal health as well as the outcome of the intrauterine pregnancy. Prenatal screening and/or diagnostic studies are necessary to evaluate the intrauterine pregnancy.

The multiple challenges in the management of a patient with HELLP syndrome, liver rupture and eclampsia.

We report a case of a patient with HELLP syndrome, hemorrhagic shock due to liver rupture and late postpartum eclampsia superimposed on lupus nephropathy and chronic hypertension. This patient was delivered at 26 weeks by C-section. Aggressive and complex surgical and medical treatments were necessary to achieve hemostasis and stabilize the patient. She recovered and was discharged to home on the 24th postpartum day in good condition. The premature baby was discharged home in fair condition on the 131st hospital day.

Prolonged in vivo residence times of antibody fragments associated with albumin.

Antibody fragments can be expressed at a high level in microbial systems, but they may have limited therapeutic value because they are rapidly eliminated from the body. We demonstrate here that site-specific conjugation or binding of bacterially derived Fab' to the long-lived protein serum albumin allows full retention of the antibody's binding characteristics while imparting the albumin's longevity in vivo. In rats the area under the curve for Fab' conjugated to rat serum albumin was 17-fold greater than for the control of Fab' conjugated to cysteine. Again, a bispecific F(ab')(2) with specificity for rat serum albumin showed an area under the curve about 8-fold greater than did a F(ab')(2) without specificity to albumin. Genetic fusions of scFv to albumin were similarly long-lived and could be expressed in yeast to provide the basis of a cost-effective production system.

A survey of pregnant women with tuberculosis at the Port Moresby General Hospital.

From March 1995 to February 1998, 110 patients diagnosed with tuberculosis (TB) in pregnancy or the puerperium at the Port Moresby General Hospital (PMGH) were surveyed. 96% were diagnosed as a result of the symptoms of tuberculosis, 4% through contact tracing. 11 of 40 patients who first attended antenatal clinic in the second trimester were not diagnosed until after delivery. The mean birthweight of term infants of TB patients was significantly less than term infants in a previous survey at PMGH. 45% of babies were growth restricted. With increasing duration of treatment, both increasing maternal weight gain in pregnancy and higher mean birthweight were found. Maternal and perinatal mortality were high in the study patients. There were 6 maternal deaths and a perinatal mortality rate of 137/1000. The majority of maternal and fetal losses occurred in patients who had pulmonary, miliary and meningeal TB. Improvement in the detection of tuberculosis in antenatal patients and the introduction of adequate treatment before delivery should prevent maternal deaths and perinatal morbidity and mortality.

Antenatal haemoglobin profile at the Port Moresby General Hospital.

From 1 November 1995 to 10 April 1996, 1001 consecutive mothers who had attended the Port Moresby General Hospital (PMGH) antenatal clinic and were delivered in the Maternity Unit of the PMGH were surveyed for their antenatal haemoglobin (Hb) levels. One or more Hb results had been noted in the antenatal records of 997 subjects. The mean of the first Hb level for this sample was 10.6 +/- 1.72 g/dl. The first Hb level was below 10.0 g/dl in 33% of the 997 subjects. The dates were reliable in 777 subjects. The mean first Hb for this group was the same as for the total sample. A subgroup of 449 subjects with reliable dates and first Hb measurement at < or = 26 weeks gestation had a first Hb mean of 10.9 +/- 1.78 g/dl. Among those with reliable dates, 569 had two or more Hb measurements, the last being at > or = 34 weeks. In this subgroup, the mean of the last Hb was 11.1 +/- 1.60 g/dl. The first Hb was measured at < or = 26 weeks in 352 subjects in this subgroup. The means for the first and last Hb levels for this subset were 10.9 +/- 1.78 and 11.1 +/- 1.57 g/dl, respectively; the paired t test did not show any difference between the first and last Hb levels. For subjects in the total sample who had two or more Hb measurements, the mean for the lowest Hb level was 10.0 +/- 1.54; 45% had levels below 10.0 g/dl at some time during the pregnancy.

Gonadotropin-releasing hormone neural projections to the systemic vasculature during sexual maturation and delayed puberty in the male Djungarian hamster.

To study GnRH neuron projections to the systemic circulation during sexual maturation and in photoperiod-induced delayed puberty, we injected fluorogold (i.p.) into long- or short-day-reared male Djungarian hamsters. Five days after fluorogold treatment, at 15, 25, or 40 days of age, most GnRH neurons were found to access the systemic vasculature from terminal fields in the median eminence and organum vasculosum of the lamina terminalis. More than 80% of GnRH cell bodies in the medial preoptic area and diagonal band of Broca contained fluorogold, the locus for the majority of the GnRH neuron system in this species. GnRH cell numbers (approximately 180) and percentages with fluorogold were the same in 15- and 25-day-old males in long days, respectively pre-and peripubertal, and were not different from that in short-day hamsters with delayed puberty. Neither the number of fluorogold-labeled GnRH somata nor intensity of the fluorogold per cell was altered in hamsters during the critical period for initiation of reproductive maturation. Only later, at age 40 days, was the percentage of GnRH somata with fluorogold increased in long-day hamsters or reduced in 40-day-old males in short days relative to that in males at 15 days of age (same day length). The findings indicate that GnRH neuron projections to regions outside the blood-brain barrier are established before puberty and are independent of photoperiod control of reproductive development.

Familial ovarian cancer: report of ovarian carcinoma in three sisters.

This is a report of ovarian carcinoma occurring in two sisters diagnosed almost at the same time, prompting prophylactic oophorectomy in a third sister. Histology of the overtly normal ovary in the third sister showed a focus of ovarian cancer. Discussion and a review of the literature suggest that any program designed to reduce the incidence of late-stage ovarian carcinoma should include the surveillance of family members of the index case, including the performance of prophylactic oophorectomy in the unaffected members of the family after they have completed their families.

Vocim--a system for measuring the larynx endoscopically.

Examination of the larynx with a flexible endoscope has been a standard procedure for over 20 years. Although excellent images have been obtained there has not been a method for achieving any objective measurements and there are clearly both research and clinical applications for this. The reason for this is firstly because of the inherent distortion of the lens and secondly because of an absence of a reliable method of assessing the relative position of the endoscope within the larynx. This paper illustrates the theory of how these problems have been overcome and describes the equipment used in the clinical situation. The first clinical trial of the Vocim system is detailed, demonstrating an error of only 4%.

Development and application of a fluorometric assay for mammalian membrane dipeptidase.

Membrane dipeptidase (EC 3.4.13.19) is a widely distributed mammalian cell surface enzyme involved in the metabolism of glutathione, leukotriene D4, and certain beta-lactam antibiotics. In this study we have developed a sensitive and rapid assay for membrane dipeptidase based on the fluorometric detection of the D-Phe released from the model substrate Gly-D-Phe. The released D-Phe is first acted on by D-amino acid oxidase in the presence of flavin adenine dinucleotide. The resulting hydrogen peroxide is then metabolized by peroxidase in the presence of the acceptor substrate p-hydroxyphenylacetic acid which is converted to a highly fluorescent compound. The assay configuration is sensitive down to 0.1 nmol D-Phe and can accurately measure membrane dipeptidase activity even in the presence of large amounts of contaminating protein. The membrane dipeptidase assay and the subsequent fluorometric detection of the released D-Phe can be performed in microtiter plates, thus taking less than 1 h to process 96 samples. This sensitive and rapid assay will be useful for the routine measurement of membrane dipeptidase activity in a number of different applications.

Orientation acuity for sine-wave gratings with random variation of spatial frequency.

Orientation discrimination thresholds were estimated for sine-wave grating stimuli using a two-alternative forced-choice procedure on the two principal, and the two main oblique axes. The two discriminanda within any trial were subject to independent random variation of their spatial frequency. The variation of spatial frequency was defined by a Gaussian probability density function. Thresholds were obtained for five different depths of frequency variation within the range 0.0-1.0 octave. We find that orientation acuity on any of the axes tested is unaffected by variation in the spatial frequency of the targets. The data support the hypothesis that orientation and spatial frequency are independent and orthogonal neural codes.

Identification and developmental expression of a novel embryonic myosin heavy-chain gene in chicken.

The developmental expression of an embryonic chicken myosin heavy-chain (MHC) gene homologous to the genomic clone pCM4.1 was examined by S1 analysis. Transcripts homologous to pCM4.1 are first detected at day 12 in ovo, and are maximally expressed between days 15-17 in ovo. No pCM4.1 transcripts are detected at earlier stages of embryogenesis or at high levels in posthatch stages. This unique pattern of expression has led to the proposal that pCM4.1 represents a previously uncharacterized MHC gene, which is confined in its expression to late embryogenesis. Genomic hybridization data, in addition to a comparison between the DNA and amino acid sequences of pCM4.1 and other characterized chicken MHC 3' end clones, provide further evidence for this proposal. We also present observations made during the sequence analysis of pCM4.1 that may be relevant to our understanding of the 3'-end processing of homologous primary transcripts, and of the mechanism controlling developmental MHC isoform transitions.

A long polypyrimidine/polypurine tract induces an altered DNA conformation on the 3' coding region of the adjacent myosin heavy chain gene.

A long (147 base pairs), natural A.T rich polypyrimidine/polypurine tract has been found 55 base pairs downstream of a chicken embryonic myosin heavy chain (MHC) gene. Analysis at the nucleotide level of nicks induced by S1 and Neurospora crassa nucleases indicate that this long interrupted polypyrimidine/polypurine tract exists in an alternate DNA structure in vitro at pH 4.5 and pH 7.5 in both supercoiled and linear plasmid DNA. The polypyrimidine/polypurine tract induces this alternate structure upon at least 200 base pairs of its 5' flanking DNA, and thus extends into the 3' coding and non-coding regions of the neighboring MHC gene. The different nicking patterns induced by the nucleases S1 and N. crassa on each strand of this alternate structure suggests that the polypyrimidine/polypurine tract may form heteronomous DNA. When this long polypyrimidine/polypurine tract is present in a supercoiled plasmid at low pH, a new and as yet undefined S1 hypersensitive DNA alteration was detected near the center of this tract.

The localization of a tcRNA102 gene near the 3' OH terminus of a fast myosin heavy chain gene. A comparison between normal and dystrophic chickens.

Two genomic fragments were isolated from a normal and a dystrophic library containing the 3'OH terminus of the fast isoform of myosin heavy chain gene. Restriction map analysis confirmed that the genes were similar. The sequences coding for myosin were defined and shown to be the same in each genomic fragment. However, using a cDNA clone for tcRNA102 and two specific oligomers for tcRNA102 sequences, we determined that only the genomic fragment from normal chick contained homologous sequences to tcRNA102. Dystrophic chick DNA did not contain these regions of homology. In addition, the normal genomic fragment transcribes tcRNA102 in vitro via RNA polymerase III while the corresponding fragment of DNA from dystrophic chick was inactive. These results suggest that there are detectable differences between the normal and dystrophic genomes in this regard.