Molecular and biochemical mechanisms ofPasteurella haemolyticaleukotoxin-induced cell death

Pasteurella haemolyticaleukotoxin (LKT) is a member of the RTX family of pore-forming toxins that kill bovine immune cells. Several studies have suggested that RTX toxins kill target cells by the induction of apoptosis. In the present study, BL3 bovine leukaemia cells were exposed to LKT and assessed by molecular and flow cytometric techniques that measure different aspects of apoptotic cell death. The intoxicated cells demonstrated morphological, light scatter and Hoechst 33258 staining characteristics consistent with cells undergoing apoptosis. The cells also exhibited internucleosomal DNA fragmentation and poly (ADP-ribose) polymerase (PARP) cleavage, both indicators of apoptosis. LKT-treated cells bound annexin-V-FITC indicating that phosphatidylserine groups were translocated from the inner to the outer leaflet of the cell membrane. The effect of LKT on cells was dose dependent and inhibitable by incubation with anti-LKT monoclonal antibody. Finally, an early step for induction of apoptosis appears to be the binding of LKT to a β2 integrin since pre-incubating cells with anti-β2 integrin antibodies inhibited LKT-induced apoptosis. This study provides new insights into understanding the pathogenesis of bovine pasteurellosis and could lead to the development of both preventative and therapeutic strategies for disease management.     

Innate immunity of surfactant protein A and D in urinary tract infection with uropathogenic Escherichia coli

Objective To investigate the role of surfactant protein (SP) - A and SP - D in urinary tract infection mouse model, and evaluate the effects of SP-A and SP-D absence on urinary tract infection. Methods SP-A and SP-D double knockout (SP-A/D KO) mice were made. SP-A/D KO and wild-type (WT) C57BL/6 female mice were used for this study. The expression of SP-A and SP-D in kidney was detected by immunohistochemistry (IHC). The levels of p - p38 and p38 protein in kidneys were measured by Western blotting. Uropathogenic Escherichia coli or buffer was delivered into the bladder of female mice. At 24 and 48 h after inoculation, CFU of Escherichia coli in the kidney and urine of the treated and control mice were measured. Histological, cellular and molecular analysis were performed by several methods of H/E staining, IHC and Western blotting. The effects of SP-A and SP-D on bacterial growth were studied in vitro. Results SP-A and SP-D in kidney were located in the proximal tubules and collecting tubules. Compared with WT mice, infected SP - A/D KO mice with UPEC had higher CFU in kidneys and urine at 24 h and 48 h, increased inflammatory cells infiltration in kidneys（P＜0.05）. Compared with WT mice, SP - A/D KO mice had higher p38 MAPK phosphorylation levels in kidneys（P＜0.05）. Growth of Escherichia coli was greatly inhibited by both SP-A and SP-D（P＜0.05）. Conclusions Both SP-A and SP-D are expressed in kidney. SP-A and SP-D can attenuate UTI induced by UPEC which may be through inhibiting bacterial growth and modulating renal inflammation.     

Embryonic heart rates before the seventh week of pregnancy

Previous studies of normal human embryonic heart rates showed an increase in the rates between the 7th and 9th gestational weeks; the rates gradually declined thereafter until the 15th week. The present study added new information about embryonic heart rates before the seventh week. The heart rates are slower than those reported to occur after the seventh week. These observations are connected with morphologic and physiologic changes during this period of embryonic life.     

Clinical experiences with fetal heart rate monitoring of one thousand patients in labor

A series of 1,000 labor patients has been monitored electronically, and their records reviewed. Monitoring provides accurate objective evidence that fetal condition is good and that tolerance for labor is normal. With a normal record, the likelihood is strong that a normal baby will be delivered. Electronic fetal heart rate monitoring is unreliable in the prediction of babies born in poor condition. It does, however, provide an excellent screening test for selection of patients for scalp pH determinations. Electronic monitoring of fetal heart rate provides early evidence of fetal anoxia. It should be carried out in all patients receiving oxytocin during labor to avoid episodes of fetal anoxia. External monitoring methods currently available permit routine monitoring of all patients in labor. Whether monitoring all normal patients will improve fetal health and survival is still an unanswered question.     

Cytopathic effects of Actinobacillus actinomycetemcomitans on monkey blood leukocytes

The suitability of cynomolgus monkeys (Macaca fascicularis) for studies concerned with the biologic properties of Actinobacillus actinomycetemcomitans is the subject of the present investigation. We found that normal monkeys harbored leukotoxic strains of A. actinomycetemcomitans in subgingival plaque samples. Monkey peripheral blood PMNs and monocytes were killed following in vitro exposure to sonic extracts of leukotoxic strains of A. actinomycetemcomitans . Monkey sera were capable of inhibiting the leukotoxic properties of A. actinomycetemcomitans sonic extracts due to the presence of IgG antibodies which neutralized the leukotoxin. Similarly, sera from patients with juvenile periodontitis (but not normal human sera) abolished leukotoxin-mediated killing of monkey PMNs. Monkey peripheral blood lymphoid cells were not killed by A. actinomycetemcomitans but demonstrated depressed responses to mitogens following pre-incubation with A. actinomycetemcomitans sonicates prepared from either leukotoxic or "non-leukotoxic" human strains. These studies suggest that cynomolgus monkeys may serve as a suitable in vitro and in vivo model for delineating more about host- A. actinomycetemcomitans interrelationships in the etiology of human periodontal disease.     

Failure of vitamin B6 to reverse the l-dopa effect in patients on a dopa decarboxylase inhibitor

Seven patients with Parkinsonism previously on l-dopa were placed on a regimen of l-dopa and alpha methyl dopa hydrazine (a dopa decarboxylase inhibitor). Two of these patients had previously shown marked clinical deterioration of the l-dopa improvement when given pyridoxine. None of the seven patients receiving alpha methyl dopa hydrazine demonstrated any change in their condition when given pyridoxine. The failure of vitamin B₆ to reverse the clinical effect of l-dopa in patients receiving both l-dopa and a peripheral dopa decarboxylase inhibitor suggests that reversal of the l-dopa effect induced by vitamin B₆ is due to increasing the activity of the enzyme dopa decarboxylase outside the central nervous system.     

Intractable paroxysmal sneezing: a conversion reaction of adolescence

A case of recurrent paroxysmal sneezing in an adolescent girl is reported with a review of the literature. The mechanisms of sneezing are described. Most cases appear to have a psychogenic etiology. We postulate that this is a conversion symptom in which the patient received secondary gain and probably represents a pre-oedipal conflict.     

Routine electronic monitoring of fetal heart rate and uterine activity during labor.

Routine electronic monitoring has been performed on 2411 labor patients at Booth Memorial Medical Center. Most recently, 88% of all patients delivered were monitored. As a direct result of this program, intrapartum stillbirths have been dramatically reduced from 1.2/1000 livebirths to 0.5/1000 livebirths. Perinatal mortality for fetuses over 1000 g has fallen to 8.8/1000 deliveries. Apgar scores below 6 at 5 minutes have decreased from a rate of 24/1000 to 14/1000 livebirths. No increase in cesarean sections for fetal distress has occurred although the primary cesarean section rate has increased over the past 10 years, apparently unrelated to fetal monitoring. It is strongly recommended that all patients in labor be monitored by currently available technics.