Effect of Doublesynch and Estradoublesynch protocols on estrus induction, conception rate, plasma progesterone, protein, and cholesterol profile in anestrus Gir heifers.

AIM: This study aimed to evaluate the efficacy of Doublesynch and Estradoublesynch protocols on estrus induction, conception rates, plasma progesterone, protein, and cholesterol profile in anestrus Gir heifers. MATERIALS AND METHODS: In this study, 50 pubertal anestrus Gir heifers were selected from the field and farm conditions. The heifers were dewormed (injection ivermectin, 100 mg, s/c) and supplemented with minerals and vitamins (injection organic phosphorus 800 mg and injection Vitamin AD3E and Biotin 10 ml i/m) and multi-mineral bolus at 1 bolus daily for 7 days. The heifers were randomly divided into three groups: Doublesynch (n=20), Estradoublesynch (n=20), and control (n=10). The animals were monitored for estrus response, estrus interval, behavioral signs, and conception rates after induced/first, second, and third cycle post-treatment. Blood samples were obtained on day 0, day 9, day 12, and on day 12 post-artificial insemination (AI) for determination of plasma progesterone, protein, and cholesterol profile. RESULTS: The estrus response rate between Doublesynch and Estradoublesynch protocols was similar between treated heifers (85% and 95%). The interval from the second prostaglandin F2α (PGF2α) injection to estrus induction did not differ between the groups (63.87±4.19 vs. 58.27±3.83 h). The conception rates following induced estrus (20% vs. 30%), at the second cycle (23.07% vs. 16.66%), at the third cycle (22.22% vs. 30.00%), and the overall conception rate (45% and 55%) within 27.89±5.75 and 26.45±5.48 days were the same across the treatment groups. The mean plasma progesterone concentrations were significantly (p<0.01) higher on day 9 (second PGF2α injection) and day 12 post-AI compared to day 0 (first PGF2α injection) and the day of fixed-timed artificial insemination. The concentrations were also significantly (p<0.05) higher in conceived than non-conceived heifers on day 9 of treatment and day 12 post-AI in both the protocols. The mean plasma cholesterol concentrations were significantly higher during peak follicular and luteal phases compared to the initial anestrus phase in both the protocols. The values were also higher in non-conceived than conceived animals in both the protocols. The plasma protein profile was not influenced by the sampling days or conceived and non-conceived status. CONCLUSION: The results showed that both Doublesynch and Estradoublesynch protocols resulted in similar estrus induction and conception rates with modulation of plasma progesterone and cholesterol profile in anestrus Gir heifers.

Effect of peripartum nutritional management on plasma profile of steroid hormones, metabolites, and postpartum fertility in buffaloes.

AIM: The aim of this study was to evaluate the influence of peripartum protein and minerals supplementation on plasma profile of steroid hormones, metabolites, and fertility in rural buffaloes. MATERIAL AND METHODS: A total of 85 advanced pregnant (~8 months) pluriparous buffaloes selected at farmers' doorstep in three tribal villages of Middle Gujarat were randomly divided into two groups, viz., control (n=45) and nutrients treatment (40). The buffaloes of treatment group (n=40), in addition to farmers feeding schedule/control, received daily 1.5 kg compound concentrate mixture (22% CP) and 50 g of chelated ASMM for 2 months each pre- and post-partum. Further, 15 buffaloes, each of control and treatment group, were injected parentrally (deep i/m) with 5 ml of micro-minerals (each ml containing Se, Zn, Cu and Mn at 5, 40, 15 and 10 mg, respectively), twice 2 months before and on the day of calving, keeping rest of the animals (control, n=30 and treatment, n=25) as controls. Blood sampling was done on days -60, -30, -15, 0, 15, 30, 45, and 60 peripartum for estimation of plasma progesterone and estradiol by standard RIA techniques and other metabolites using assay kits on biochemistry analyzer. The puerperal events and postpartum fertility were monitored through history and by fortnightly palpation per rectum till day 45 and then again at 120 days postpartum for both the groups and subgroups. RESULTS: The mean plasma progesterone concentrations in all groups declined significantly (p<0.05) from day 60 to day 15 prepartum, reached to the basal levels (<0.5 ng/ml) on the day of parturition, and subsequently, reduced nonsignificantly till day 15 postpartum and then showed a rising trend from day 30 to 60 postpartum with significantly higher values at day 45 and/or 60. The mean plasma estradiol values increased with approaching parturition and were at its peak on the day of calving (p<0.01). Thereafter, there was a rapid fall in the levels by day 15 and it remained low till day 45-60 postpartum. The blood glucose values showed an increasing trend with advancing gestation, reaching the highest on the day of calving, dropped significantly (p<0.01) within 15 days postpartum, and thereafter showed consistent values. The buffaloes supplemented with peripartum nutrients maintained significantly (p<0.05) higher blood glucose concentrations than the control during the peak lactation. The plasma protein levels varied significantly (p<0.05) between days within the group with the lowest values on the day of calving, as well as between groups with higher (p<0.05) values on day 30 and 60 postpartum in treated group. Micro-minerals injected did not reveal significant influence on steroid hormones, blood glucose, or plasma protein. The mean plasma total cholesterol was significantly lower (p<0.05) in treatment than the control group. The mean values in micro-minerals injected subgroup were higher than the non-injected control subgroup during postpartum phase. The mean plasma triglyceride values in the pregnant buffaloes under both the groups and subgroups gradually decreased as parturition approached with significantly lowest values on the day of calving. The values increased nonsignificantly by day 15 and then remained steady throughout postpartum period without influence of nutrient supplementation or micro-minerals injection. The incidence of retained fetal membranes (RFMs) was 5.00 and 13.33% in treatment and control groups, respectively, with placental expulsion time of 3.27±0.37 and 4.44±0.53 h (p>0.05). The micro-minerals injection appreciably reduced the incidence of RFMs and significantly (p<0.05) reduced the placental expulsion time over non-injected controls. In treatment group, the period for involution of uterus was significantly shorter (29.39±0.50 vs. 32.12±0.82 days, p<0.05), with early onset of first postpartum estrus (67.65±1.67 vs. 79.43±3.06 days, p<0.01), shorter service period (90.89±4.41 vs. 105.09±4.76 days, p<0.05) and higher conception rate (55.00 vs. 40.00%) than in control group. The micro-minerals injection apparently and/or significantly improved all these traits in both the groups. Thus, the postpartum reproductive performance was significantly improved in treated than control groups and subgroups. CONCLUSION: The results showed that nutrient supplementation in terms of high protein concentrate, ASMM and injection of sustained release micro-minerals (Se, Zn, Cu, and Mn) during transition period minutely altered the plasma steroid hormones and blood metabolites though it significantly improved the postpartum reproductive performance in buffaloes under field conditions.

Are there differences in outcome after elective sigmoidectomy for diverticular disease and for cancer? A national inpatient study.

AIM: The postoperative outcome after elective sigmoidectomy for diverticulitis has not been compared to that for cancer. The study aimed to evaluate the differences in the postoperative outcome after sigmoidectomy for diverticular disease and cancer. METHOD: The National Inpatient Sample Database was used to identify patients who underwent elective sigmoid resection for diverticular disease or cancer between 2004 and 2011. After excluding patients with metastatic cancer and preoperative weight loss, sigmoid cancer and diverticulitis patients were matched using propensity score, controlling for age, gender, race, type of operation (open vs laparoscopic) and comorbidities. The end-points of interest were infective complications, reoperation, anastomotic leakage, rebleeding, length of hospital stay and in-hospital mortality. RESULTS: After propensity score matching (diverticulitis 11 192 patients, sigmoid cancer 11 192 patients), the mean age was 65 ± 12.5 years, 53.8% were male and 61.5% were Caucasian. Only 18.0% of the operations were done by laparoscopy. The overall complication rate was 17.7% and the in-hospital mortality rate was 0.9%. The diverticulitis group had a higher rate of surgical site infection (3.2% vs 2.6%, P = 0.004), intra-abdominal abscess formation (1.2% vs 0.4%, P < 0.0001) and reoperation (6.1% vs 4.1%, P < 0.0001) compared with the cancer group. The cancer group had a higher incidence of pneumonia (1.9% vs 1.5%, P = 0.01) and anastomotic leakage (9.2% vs 8.3%, P = 0.001). There was no difference in sepsis, deep vein thrombosis, respiratory failure, renal failure, rebleeding, overall complication rate or length of hospital stay. Subgroup analysis showed a higher in-hospital mortality for cancer than for diverticulitis patients whether resected by open or by laparoscopic surgery. CONCLUSION: Although elective sigmoidectomy for diverticular disease has a higher risk of infective complications, elective sigmoidectomy for cancer has a higher risk of anastomotic leakage.

Comparative efficacy of different estrus synchronization protocols on estrus induction response, fertility and plasma progesterone and biochemical profile in crossbred anestrus cows.

AIM: To evaluate estrus induction response and fertility including plasma progesterone and biochemical profile following use of three standard hormonal protocols in anestrus crossbred cows. MATERIALS AND METHODS: The study was carried out on 40 true anestrus and 10 normal cyclic cows. 10 anestrus cows each were treated with standard intravaginal controlled internal drug release (CIDR) device, Ovsynch (GPG) protocol, and Norgestomet ear implant with fixed-time artificial insemination (FTAI). 10 anestrus cows were kept as untreated control while 10 cows exhibiting the first estrus within 90 days postpartum without any treatment served as normal cyclic control. Blood samples were obtained from treated cows on day 0, 7, 9 (AI) of treatment and day 21 post-AI, and from control groups on the day of AI and day 21 post-AI for estimation of plasma progesterone, protein, cholesterol, calcium, and inorganic phosphorus profile. RESULTS: The use of CIDR, Ovsynch, and Norgestomet ear implant protocols resulted in 100% estrus induction with conception rates at induced estrus of 60%, 50%, and 50%, and the overall of three cycles as 80%, 80%, and 70%. In untreated anestrus control (n=10), only three cows exhibited spontaneous estrus within 90 days of follow-up and conceived giving the first service and overall conception rates of 66.66% and 30.00%, respectively. In normal cyclic control (n=10), the conception rates at first and overall of three cycles were 50% and 80%. The overall mean plasma progesterone (P4) concentrations in anestrus cows studied on day 0 (initiation), 7 (prostaglandin injection and/or removal of implant), 9 (FTAI) of treatment and on day 21 post-AI revealed that the values on day 7 and 21 were significantly (p<0.01) higher than other two periods in all three groups. The concentrations were significantly (p<0.05) higher in conceived than non-conceived group on day 21 post-AI in CIDR (4.36±0.12 vs. 1.65±0.82 ng/ml) and Ovsynch (4.85±0.62 vs. 1.59±0.34 ng/ml), but not in Norgestomet ear implant (4.50±0.53 vs. 3.02±1.15 ng/ml) or normal cyclic group (5.39±0.67 vs. 3.13±0.37 ng/ml). The cholesterol and protein levels were significantly higher, but not the calcium and phosphorus, in normal cyclic control than in anestrus groups. The influence of treatment days and pregnancy status was not significant for any of the biochemical constituents in any of the groups. CONCLUSION: Ovsynch and/or CIDR synchronization protocol can be effectively used to improve fertility up to 80% in anestrus cows, as compared to 30% in anestrus control, combined with plasma progesterone to delineate the reproductive status before and after treatment.

Relative efficacy of egg yolk and soya milk-based extenders for cryopreservation (-196°C) of buffalo semen.

AIM: The aim was to compare commercially available soybean milk-based extenders, viz. Bioxcell(®) and Optixcell(®) (IMV, France) with standard Tris-citrate-fructose-egg yolk-glycerol (TFYG) extender for cryopreservation of buffalo semen. MATERIALS AND METHODS: Semen was collected twice a week in artificial vagina from six sexually mature, 4-6 years old, healthy breeding bulls of Surti buffalo breed. In all 48 qualifying ejaculates (8 per bull) having initial motility >70% were split into three equal aliquots and were diluted (at 34°C keeping 100×10(6) sperm ml(-1)) in TFYG, Bioxcell and Optixcell extenders. The French mini straws filled from each aliquot were gradually cooled to 4-5°C, equilibrated at 4°C for 4 h and frozen in liquid nitrogen 2 vapor using programmable biofreezer. Just before freezing (post-equilibration) and 24 h after frozen storage, the samples were evaluated for various sperm quality parameters using standard protocols. Frozen semen straws were thawed in a water bath at 37°C for 30 s. The post-thaw incubation survival (37°C for 1 h) was assessed through motility rating at 0, 30 and 60 min of incubation. RESULTS: The mean percentages of prefreeze sperms in TFYG, Bioxcell and Optixcell extenders in terms of progressive motility (69.48±0.37, 68.02±0.49, 70.94±0.38), viability (79.21±0.39, 77.38±0.48, 81.58±0.38), total abnormalities (7.90±0.14, 8.60±0.16, 7.08±0.15), intact acrosome (89.54± 0.18, 88.58±0.22, 90.52±0.21) and hypoosmotic swelling (HOS) reactivity (67.96±0.32, 65.65±0.42, 70.23±0.37) varied significantly (p<0.05) between extenders. Similar pattern of significant (p<0.05) variations between these extenders for post-thaw sperm progressive motility (47.71±0.79, 44.38±0.85, 49.90±0.90), viability (57.19±0.79, 53.85±0.84, 59.67±0.91), total abnormalities (12.33±0.17, 12.75±0.21, 11.27±0.18), intact acrosome (76.83±0.23, 75.90± 0.27, 78.50±0.25) and HOS reactivity (45.02±0.84, 42.31±0.82, 47.81±0.90) was also observed for TFYG, Bioxcell and Optixcell extenders. The recently launched improved soybean milk-based extender Optixcell excelled the older Bioxcell extender and even standard TFYG in respect of some of the sperm quality parameters. CONCLUSION: The advantages of soy lecithin-based bovine semen extenders over egg yolk regarding sanitary issues are unquestionable but still egg yolk-based semen extenders are widely used because of the cost factor and good in vivo fertility results.

Depression in the elderly in Vellore, South India: the use of a two-question screen.

BACKGROUND: Depression in the elderly is a common and disabling condition. The aim of the study was to evaluate the sensitivity and specificity of a two-question screen to identify depression and common mental disorders in the elderly. METHOD: Residents of a ward in the town of Vellore were identified by a door-to-door survey from which 204 subjects aged over 60 years were selected for the study by systematic random sampling. They were screened using the two-question screen. The Revised Clinical Interview Schedule (CIS-R) was employed to confirm the diagnosis. RESULTS: The prevalence of depression and common mental disorder, using the CIS-R standard, was found to be 31.5%. The two-question screen has a sensitivity of 93.8% and specificity of 48.2%. CONCLUSIONS: The high sensitivity of the two-question screen makes it a useful screening method which can be employed by health workers in the field.

Accurate axillary nodal staging can be achieved after neoadjuvant therapy for locally advanced breast cancer.

Lymph node status remains the most important prognostic indicator for breast cancer. Recent reports have established that the accuracy of assessing lymph node status is proportional to the number of nodes dissected. The accuracy of axillary staging following neoadjuvant chemotherapy has been cited as a technical concern due to limited node retrieval. The current study attempts to evaluate the ability to perform sentinel node biopsy (SNB) and formal axillary node dissection (AND) following neoadjuvant chemotherapy and to compare these results with non-neoadjuvant patients. One hundred sixteen consecutive patients undergoing SNB with simultaneous AND were retrospectively reviewed. Forty-two of these patients were treated with neoadjuvant chemotherapy prior to AND. Overall success rate in performing SNB in the neoadjuvant group was 95 per cent, and no false negatives have been noted to date. The overall SNB success rate in the non-neoadjuvant group was also 95 per cent with a false negative rate of 3 per cent. After AND in each group, a mean of 21 nodes were retrieved in the neoadjuvant group and 17.9 nodes in the non-neoadjuvant group (P = 0.018). In the neoadjuvant group, there were 19 node positive patients (42%) and 21 patients (28%) in the non-neoadjuvant group (P = 0.16). The mean number of positive nodes per patient was also similar between the two groups (2.9 in the neoadjuvant group vs 1.67 in the non-neoadjuvant group, P = 0.10). Following neoadjuvant therapy, accurate evaluation of the axilla is feasible. In this study, the mean number of nodes is significantly different in favor of the neoadjuvant group, but there is no significant difference in the number of node positive patients identified or in the mean number of positive nodes identified per patient. SNB is technically feasible with accuracy similar to that seen in patients with no history of neoadjuvant therapy. Neoadjuvant chemotherapy extends the use of breast-conserving therapy without sacrificing the ability to accurately stage the axilla either by use of standard axillary dissection or SNB.

Neoadjuvant chemotherapy for locally advanced breast cancer results in alterations in preoperative tumor marker status.

Neoadjuvant therapy followed by breast-conserving surgery has become an acceptable option for patients with locally advanced breast cancer. Although a distinct survival benefit has not been demonstrated using this approach, several questions have been raised following such therapy including its effects on receptor status and tumor markers. The current study retrospectively reviews estrogen receptor (ER), progesterone receptor (PR), and HER2-neu status in 55 consecutive patients treated by neoadjuvant chemotherapy. Preoperative and postoperative tumor markers were available for 43 of the 55 patients (78%). The pathologic complete tumor response rate (pCR) for this group was 19 per cent (8/43). Of those patients who did not achieve a pCR (n = 35), a change in tumor markers was seen in 25.7 per cent (9/35) of patients. When compared to a control group not undergoing neoadjuvant therapy, a significantly higher percent change in marker expression was noted in the neoadjuvant group (25.7% vs 5.9%, P = 0.046). ER, PR, and HER2-neu status remain important prognostic indicators for breast cancer. Tumor markers are useful in planning adjuvant therapy regimens. In this review, nearly 19 per cent of patients achieved a pCR. In patients not achieving a pCR, one in four patients had at least one change in tumor marker status. This study demonstrates the importance of establishing receptor and marker status prior to neoadjuvant therapy, as many patients will achieve a pCR and make tumor analysis impossible. Postoperative marker studies should be performed given the possibility of a change in status. The clinical relevance of this data will require further long-term follow-up. Until such data becomes available, caution should be considered when basing adjuvant therapy regimens on preoperative tumor marker studies alone.

Comparison of 3 vancomycin dosage regimens during hemodialysis with cellulose triacetate dialyzers: post-dialysis versus intradialytic administration.

AIMS: Traditionally, vancomycin is administered following dialysis to minimize drug loss when high-flux membranes are employed. Unfortunately, this approach is extremely inconvenient for patients and staff, requiring the patients to remain in the unit for at least 1 hour following dialysis. This study was designed to evaluate the feasibility of administering vancomycin during hemodialysis. Specifically, this study was designed to compare the pharmacokinetics of vancomycin when administered during the last 1-2 hours of dialysis (i.e. intra-dialytic administration) to that administered after completion of dialysis. MATERIALS AND METHODS: In a randomized, 3-way crossover trial, the pharmacokinetics of vancomycin were evaluated in 9 hemodialysis patients, comparing vancomycin 15 mg/kg following dialysis (Phase I), vancomycin 15 mg/kg during the last hour of hemodialysis (Phase II) or vancomycin 30 mg/kg during the last 2 hours of hemodialysis (Phase III). Vancomycin plasma concentrations were obtained over an 8-day period and subsequent comparisons between the treatment approaches were made with paired t-tests or ANOVA, as appropriate. Dialysate vancomycin concentrations determined on Day 1 and Day 3 of Phases II and III were used to calculate the fraction of vancomycin dose removed, and were compared to plasma data using paired t-tests. RESULTS: Vancomycin was significantly removed (33.4 to 39.5%) during a 3- to 4-hour high-flux dialysis session occurring on Day 3 after vancomycin administration. Mean serum concentrations immediately following intradialytic vancomycin administration of 15 mg/kg over the last hour of dialysis or 30 mg/kg over the last 2 hours of dialysis were initially high (77.7 and 95.5 mcg/ml respectively), but fell to 25.9 and 40.5 mcg/ml, respectively, by 4 hours post-dialysis. Predialysis concentrations on Days 3, 5 and 8 were similar for vancomycin 30 mg/kg administered over the last 2 hours of dialysis as compared with a 15 mg/kg dose given after dialysis. Vancomycin 15 mg/kg over the last hour of dialysis resulted in significantly lower subsequent predialysis concentrations than the other dosing schemes. CONCLUSIONS: Vancomycin administration of 30 mg/kg over the last 2 hours of dialysis achieves serum concentrations similar to conventional dosing of 15 mg/kg after dialysis and would allow dosing on a weekly basis.

Clinical manifestations and management of pediatric HIV infection.

HIV infection has emerged as a colossal problem with epidemic proportions. According to an estimate from UNAIDS about 36.1 million people all over the world are infected at present. In India about 3.5 million people are infected. The infection has evolved into phase II process of disease evolution, spreading from high-risk population to the general population. The antenatal HIV seropositivity has shown a steady increase from 0.1% to 2% in some tertiary care hospitals in Mumbai. Pediatric HIV infection presents with diverse clinical manifestations. In developing countries like India, diagnosis of infection during first year of life in perinatally exposed infants poses a problem due to lack of easy accessibility and increased cost of diagnostic facilities like HIV-PCR, CD4/CD8 counts and viral cultures. Moreover, lack of adequate drugs and exorbitant cost of sustaining antiretroviral therapy complicates the management issues. An assortment of antiretovirals is available in USA and other developed countries. In India drugs like zidovudine, lamivudine, stavudine, nevirapine and indinavir are available and are used in symptomatic patients. CDC has defined definite treatment guidelines for pediatric population recently. These guidelines need to be modified in our set up. At the present juncture in India the emphasis remains on the prevention and treatment of opportunistic infections like tuberculosis and pneumocystis carinii and on prevention of perinatal transmission with zidovudine. This brief review deals with various clinical manifestations as relevant in a developing country like India and recent advances in antiretroviral therapy.

Mother-to-child transmission of the human immunodeficiency virus in Texas.

BACKGROUND: The Pediatric Spectrum of HIV Diseases (PSD) project has been collecting data on HIV-exposed children in Texas since 1989. These data have now been analyzed to describe mother-to-child transmission in Texas and to provide much needed information on the magnitude of the pediatric HIV epidemic in the state. METHODS: We examined trends in the numbers of perinatally exposed children and perinatally acquired cases of HIV in the Texas PSD cohort. We calculated transmission rates and relative risks for 656 children born from January, 1995, to July, 1998, that received all or part of the ACTG 076 regimen. RESULTS: Only a small proportion (38%) of pairs of an HIV-infected mother and her HIV-exposed child received the full AIDS Clinical Trial Group 076 (ACTG 076) regimen; only 73% of the mothers received at least some prenatal care. In recent years, however, the numbers of perinatally exposed children and perinatally acquired cases of HIV have decreased in Texas. Univariate analyses showed that a reduction in the vertical transmission of HIV was associated with receipt of a full ACTG 076 regimen, receipt of a partial ACTG 076 regimen and residence in Dallas County. CONCLUSIONS: Findings identify a gap in meeting the health care needs of pregnant HIV-infected women and suggest missed opportunities to prevent mother-to-child transmission of HIV. At the same time this study confirms progress in prevention efforts to reduce mother-to-child transmission of HIV in Texas.

Dexamethasone affects cytokine-mediated adhesion of HL-60 human promyelocytic leukemia cells to cultured dermal microvascular endothelial cells.

Leukocyte endothelial adhesion (LEA) is the prelude to a complex cascade of reactions following an immunological challenge. Recently, LEA has been implicated in the molecular basis of several dermatological disorders. While the role of proinflammatory cytokines, such as interleukin-1 beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), in LEA has been investigated using nondermal models, limited data exist regarding their effects on LEA in dermal models. This study shows that cotreatment of cultured human dermal endothelial cells (CADMEC) with IL-1beta and TNF-alpha resulted in a marked increase in the adherence of human promyelocytic leukemia (HL-60) cells to CADMEC and an increase in expression of intercellular adhesion molecule-1 and E-selectin. Pretreatment of CADMEC with dexamethasone, a long-lasting glucocorticoid, resulted in a decrease in both HL-60 cell adhesion to CADMEC and adhesion molecule expression. Taken together, these data demonstrate that LEA may play a role in inflammatory skin conditions and in the mechanisms underlying the potential use of glucocorticoids as a treatment option.

Subjective assessment of fork-lift truck seats under laboratory conditions.

A subjective assessment of fork-lift truck seats has been carried out to assess the range of preferred seat dimensions and the acceptability of different seat adjustments to fork-lift truck drivers. Twelve fork-lift truck seats fitted to a fixed bench were assessed by twelve fork-lift truck drivers. For each seat, each driver completed a questionnaire that covered the following areas: eleven seat dimensions, four seat adjustments and other features (arm rests, safety belts and safety wings). The drivers assessed the dimensions of a seat chosen at random, before moving to the adjacent seat in either a clockwise or anticlockwise direction. Before each assessment, they were asked to look forward and backward in the seat as though they were driving a fork-lift truck. In general, significant correlations were obtained between the subjective assessments and objective measurements of the seat dimensions. This enabled preferred seat dimension ranges to be defined. Fork-lift truck drivers ranked the forward-backward and the backrest inclination adjustments as most important. The results showed that although the drivers had previous experience in the use of suspension seats, they did not understand the purpose of the weight adjustment. All adjustments should be easy to find, accessible, easy to move and they should enable a range of adjustments. In addition, clear and simple information on the seat is needed, especially for the weight adjustment. The most recently designed seats generally had dimensions within the preferred ranges compared with the older generation seats, mainly because they had more adjustments. However, the results show that adjustments on fork-lift truck seats could be improved further. Some tentative conclusions are made for the preferred fork-lift truck seat dimensions and adjustment ranges which are based on the evidence from this restricted sample of fork-lift truck drivers. They may be useful for seat and truck designers in the preparation of a standard on fork-lift truck seat dimensions.

Fimbria-mediated enhanced attachment of nontypeable Haemophilus influenzae to respiratory syncytial virus-infected respiratory epithelial cells.

Respiratory syncytial virus (RSV) infection is known to predispose children to otitis media and sinusitis due to bacteria such as nontypeable Haemophilus influenzae (NTHI). In this study, we investigated the role of NTHI surface outer membrane protein P5-homologous fimbriae (P5-fimbriae) in attachment to RSV-exposed A549 epithelial cells. Analysis by fluorescence flow cytometry showed that a live P5-fimbriated NTHI strain (NTHIF+) attached to a higher proportion of RSV-exposed A549 cells than to control cells (mean, 68% for RSV versus 29% for control; P = 0.008), while attachment of the P5-fimbriae-deficient isogenic mutant strain (NTHIF-) was significantly lower than in control cells and rose only slightly following RSV exposure (mean, 17% for RSV versus 10% for control, P = 0.229). Attachment of NTHIF+ did not correlate with the amount of RSV antigen expressed by A549 cells. Furthermore, paraformaldehyde-fixed NTHIF+ also demonstrated an enhanced binding to RSV-exposed cells. Observations by transmission electronic microscopy showed that the mean number of bacteria attached per 100 RSV-exposed A549 cells was higher for NTHIF+ than NTHIF- (99 versus 18; P < 0.001). No intracellular bacteria were identified. UV-irradiated conditioned supernatants collected from RSV-infected A549 cultures (UV-cRSV) also enhanced the attachment of NTHIF+ to A549, suggesting the presence of a preformed soluble mediator(s) in UV-cRSV that enhances the expression of receptors for P5-fimbriae on A549 cells. In summary, RSV infection significantly enhances NTHI attachment to respiratory epithelial cells. P5-fimbria is the critical appendage of NTHI that participates in this attachment. In clinical settings, blocking of the P5-fimbria-mediated attachment of NTHIF+ by passive or active immunity may reduce the morbidity due to NTHI during RSV infection.

Cytokine modulation of human corneal epithelial cell ICAM-1 (CD54) expression.

To determine whether pro-inflammatory cytokines modulate intercellular adhesion molecule-1 (ICAM-1; CD54) expression on cultured primary human corneal epithelial cells (HCEs), confluent HCEs were treated with various concentrations of interferon-gamma(IFN-gamma), interleukin-1alpha(IL-1alpha), IL-1beta, IL-4, tumor necrosis factor-alpha (TNF-alpha), or combinations over time. ICAM-1 expression was measured by flow cytometry and/or a cell-based ELISA using a monoclonal mouse anti-human CD54 antibody. The apparent MW of ICAM-1 protein was determined by immunoprecipitation of biotinylated HCEs. RT-PCR was used to detect ICAM-1 RNA. The mature cell surface form of HCE ICAM-1 was approximately 110 kDa as determined by immunoprecipitation. IFN-gammaand TNF-alpha induced both dose- and time-dependent increases in ICAM-1 expression. An approximately 20-fold increase in ICAM-1 was seen at 50-100 U IFN-gamma ml-1. ICAM-1 specific mRNA accumulated approximately 4.5-fold after IFN-gammatreatment. TNF-alpha(100 U ml-1) induced a consistent approximately 6.0-fold increase in ICAM-1 expression. When IFN-gammaand TNF-alpha were mixed, at sub-optimal concentrations of each, a synergistic effect on ICAM-1 expression was not detected. Neither IL-4, IL-1alpha nor IL-1beta affected ICAM-1 expression in a consistent fashion. In summary, ICAM-1 was modulated on primary human corneal epithelial cells by the cytokines IFN-gamma and TNF-alpha in a dose- and time-dependent fashion. Cytokine modulation of corneal epithelial cell ICAM-1 during inflammation may contribute to corneal epithelial cell injury by aiding the attachment of inflammatory cells such as eosinophils which express the receptor for ICAM-1, the beta2 integrins (CD11a,b,c/CD18).

Arachidonylethanolamide (AEA) activation of FOS proto-oncogene protein immunoreactivity in the rat brain.

It is thought that the physiological actions of endogenous cannabinoid arachidonylethanolamide (AEA), as well as exogenous cannabinoids such as Delta9-tetrahydrocannabinol (THC), are mediated by two subtypes of cannabinoid receptors, CB1 and CB2, which have recently been characterized. Injection of AEA leads to alterations in motor behavior and endocrine function. While these phenomena have been well characterized, the neuronal substrate of AEA's actions remains undetermined. In this study, FOS immunoreactivity (FOSir) was used to map rat brain nuclei that are responsive to a single intracerebroventricular injection of AEA. The results showed that FOSir was induced in several nuclei including the bed nucleus of the stria terminalis (BNST), paraventricular nucleus of the hypothalamus (PVN), central nucleus of the amygdala (Ce), periaqueductal gray area (PAG), dentate gyrus in the hippocampus (Dg), paraventricular nucleus of the thalamus (PVA), median preoptic nucleus (MnPO), periventricular nucleus (Pe), caudate putamen (CPU) and the ependymal lining of the ventricles. The pattern of activation identified correlates, in part, with the distribution of CB receptors. At the same time, a new subset of nuclei, without demonstrable CB receptors, have been shown to respond to an AEA challenge. Activation of these nuclei is consistent with the physiological effects of AEA. These findings provide valuable information on the response to AEA at the level of neuronal activation and provide the basis for a broader understanding of the possible role of CB receptors in the modulation of motor and endocrine function associated with the use of exogenous cannabinoids, such as marijuana.

Autocrine regulation and experimental modulation of interleukin-6 expression by human pulmonary epithelial cells infected with respiratory syncytial virus.

The mechanisms of regulation of interleukin-6 (IL-6) production in respiratory syncytial virus (RSV)-infected respiratory epithelial cells were evaluated in A549 cell cultures. Incubation with purified RSV resulted in significant production of IL-1alpha, IL-1beta, IL-6, and tumor necrosis factor alpha (TNF-alpha). Addition of saturating concentrations of neutralizing antibodies against IL-1alpha, IL-1beta, or TNF-alpha into purified RSV-infected cell cultures resulted in a significant inhibition of IL-6 production, although anti-IL-1alpha antibody had the most predominant effect (80% inhibition). Anti-IL-1alpha antibody also almost completely blocked the expression of mRNA for IL-6. Addition of therapeutic concentrations of dexamethasone (1 microM) or ribavirin (90 microg/ml), an antiviral agent, also significantly inhibited the synthesis of IL-6. Hence, in clinical settings, pharmacological agents such as the specific antagonists of IL-6-inducing cytokines, as well as dexamethasone and ribavirin, could be used to modulate IL-6 production.