First Report of Capsicum chlorosis virus Infecting Waxflower (Hoya calycina Schlecter) in the United States.

In February 2013, an ornamental waxflower (Hoya calycina Schlecter) with leaves displaying concentric chlorotic and necrotic rings surrounding sunken, necrotic lesions typical of tospovirus infection was observed at a community garden in Honolulu, HI. Symptomatic leaf tissue tested negative for Tomato spotted wilt virus (TSWV), a common tospovirus in Hawaii, using a TSWV ImmunoStrips (AgDia, Elkhart, IN) assay following the manufacturer's instructions. Double-stranded RNAs were isolated from a symptomatic leaf and reverse transcribed using random primers (2). The cDNA was then used as template in a universal tospovirus PCR assay using primers gL3637 and gL4435c, which amplify sequences of the L segment encoding the RNA-dependent RNA polymerase of tospoviruses (1). An ~800-bp product was amplified and cloned using pGEM-T Easy (Promega, Madison, WI). Three clones were selected and found to be identical by dye-terminator sequencing performed at the University of Hawaii's Advanced Studies in Genomics, Proteomics, and Bioinformatics laboratory. Following primer sequence trimming, the 773-bp sequence (GenBank Accession No. KF030938) was found to be 97, 88, and 87% identical to Capsicum chlorosis virus (CaCV; a tentative species in the family Bunyaviridae, genus Tospovirus) strains Ch-Har (GU199334), TwTom1 (HM021140), and AIT (DQ256124), respectively. To confirm the presence of CaCV, the cDNA was also used as template in a universal tospovirus PCR assay with primers 3'T12 and TsMCR2 which amplify a region of the S segment of tospoviruses (3). The amplification product from this assay was cloned and sequenced as described above and found to be 93 to 98% identical to CaCV nucleotide sequences present in GenBank. Attempts to detect the CaCV strain in waxflower using a watermelon silver mottle virus and groundnut bud necrosis virus triple antibody sandwich ELISA (AgDia) were unsuccessful. No other plants in the community garden had typical tospovirus-like symptoms; however, samples from tomato (Solanum lycopersicum L.; two samples), chili pepper (Capsicum spp.; four samples), eggplant (Solanum melongena L.; one sample), and passionfruit (Passiflora edulis Sims; one sample) with virus-like symptoms were collected from the garden and had RNA isolated using a NucleoSpin RNA II kit (Macherey-Nagel, Bethlehem, PA). No tospoviruses were detected in any of these samples with the RT-PCR assay using primers gL3637 and gL4435. The waxflower plant infected with CaCV was immediately removed by community garden members and destroyed, preventing any additional serological or biological assays to be performed. CaCV is transmitted by several species of thrips, including Thrips palmi, which is present in Hawaii. Waxflower is not native to Hawaii and it is unclear whether CaCV entered Hawaii in this plant or whether it was infected by viruliferous thrips. A survey for CaCV in known hosts is essential to determine the geographic distribution of CaCV in Hawaii, as this virus poses a considerable threat to tomato, chili pepper, and phalaenopsis orchid production in Hawaii and the United States. References: (1) F.-H. Chu et al. Phytopathology 91:361, 2001. (2) M. J. Melzer et al. Virus Genes 40:111, 2010. (3) M. Okuda and K. Hanada. J. Virol. Methods 96:149, 2001.

First Report of Taro vein chlorosis virus Infecting Taro (Colocasia esculenta) in the United States.

In March 2013, taro plants (Colocasia esculenta [L.] Schott cv. Iliuaua) with leaves displaying veinal chlorosis and necrosis were observed on the island of Molokai. These symptoms were similar to those of taro vein chlorosis, a disease of taro caused by Taro vein chlorosis virus (TaVCV; family Rhabdoviridae, genus Nucleorhabdovirus). To explore this possibility, RNA was isolated from both symptomatic and asymptomatic taro leaves using the NucleoSpin RNA II extraction kit (Macherey-Nagel, Bethlehem, PA) according to the provided protocol, except that RLT Buffer (Qiagen Inc., Valencia, CA) was used as the initial extraction buffer. The RNAs were converted to cDNA using random primers and MMLV-RT reverse transcriptase (Promega, Madison, WI). The cDNA underwent PCR assays using primer sets Pol2A1/Pol2A2 and Cap2A/Cap2B which target the RNA-dependent RNA polymerase (RdRp) and putative nucleocapsid genes of TaVCV, respectively (1). Amplification products of the correct size were obtained for both primer sets, and these underwent molecular cloning using pGEM-T Easy (Promega). Three clones were selected and their sequences determined by dye-terminator sequencing. After primer sequence removal, the Pol2A1/Pol2A2 product (952 bp; GenBank Accession No. KF921085) and Cap2A/Cap2B product (1,050 bp; KF921086) were found to be 79 and 84% identical to a Fijian strain of TaVCV (AY674964), respectively. Samples from 328 plants with and without taro vein chlorosis symptoms were collected from 35 sites on five of the Hawaiian islands and assayed for TaVCV using the Pol2A1/Pol2A2 primer set as described above. The incidence of TaVCV in these samples was 21.6%, with positive samples coming from each island. Although a very strong association between symptoms and the presence of TaVCV was observed, eight asymptomatic plants were also positive, suggesting the detection assay was able to detect the virus before the onset of symptoms. Conversely, three symptomatic plants were found to be negative, suggesting the Pol2A1/Pol2A2 PCR assay might not detect all strains of TaVCV in Hawaii. A digoxygenin-labeled probe (Roche Applied Science, Indianapolis, IN) derived from the Pol2A1/Pol2A2 amplification product of one sample hybridized with the cDNA of only four of nine TaVCV-infected samples collected from three different islands in a dot-blot hybridization assay performed at high stringency. This probe did not hybridize with the cDNA of five TaVCV-negative samples. TaVCV exhibits a great deal of genetic diversity in the South Pacific nations where it is found; nucleotide divergence of up to 27% in regions of the RdRp gene has been reported (1). The high genetic divergence between the TaVCV isolate characterized in Hawaii and the TaVCV accession in GenBank, as well as the dot blot hybridization assay results support this observation. The widespread distribution of TaVCV in Hawaii suggests it is not a recent introduction. However, the common practice of farmers sharing taro propagules has likely accelerated its spread. An arthropod vector of TaVCV has yet to be identified, so it is unknown whether natural spread is also occurring in Hawaii. Taro has both economic and cultural importance to Hawaii. These findings, representing the first detection of TaVCV in Hawaii and the United States, illustrate the need to develop virus-free germplasm for local, national, and international distribution of this important staple crop. Reference: (1) P. Revill et al. J. Gen Virol. 86:491, 2005.

The role of the CNM in newborn management.

This article provides a review of the physical, neurologic, and gestational age assessment of the newborn. Examination techniques, normal findings, and common deviations from normal are included. Nurse-midwives do not always manage the care of the neonate beyond the first few weeks of life, but they are responsible for newborn screening, provision of anticipatory guidance, and referral. Common concerns of parents in the neonatal period are feeding, elimination, crying, and sleep patterns. In addition, the nurse-midwife should provide information about immunizations, accident prevention, and dental health.

Premenstrual syndrome. Current concepts in diagnosis and management.

Premenstrual syndrome (PMS), a constellation of physical and psychologic symptoms, is a common complaint among women of reproductive age, particularly women in their 30s or older and among women of greater parity. Symptoms must occur in the luteal phase with resolution or improvement within one or two days of the onset of menses. Numerous hypotheses have been espoused to explain the etiology of PMS, but no definitive cause has been established. The management of PMS may include simple measures such as exercise, dietary changes, and vitamin supplementation. Pharmacologic agents such as progesterone, prostaglandin inhibitors, diuretics, oral contraceptives, or psychotropic drugs may be useful in selected clients. Support and counseling is of paramount importance in the management of PMS. Nurse-midwives can manage women who have minor and transient physiologic and emotional changes associated with mild degrees of PMS. Women with moderate and severe symptoms should be referred for evaluation and follow-up.

[Effects of nine prescriptions on fever in rats induced by endotoxin].

In this paper in is reported that nine prescriptions often used in traditional Chinese medicine for the treatment of diseases with fever were tested experimentally for the effect on fever of rats induced by endotoxin. The results are as follows: Da-yuan-decoction, Gan-lu-xiao-du-pill, Lian-pu-decoction, Cai-hao-mixture, Da-huang-huang-lian-jie-du-decoction, Tao-ren-cheng-qi-decorction, Ge-gen-qin-lian-decoction, Yin-qiao-jie-du-decoction etc. can notably reduce fever in rats induced by endotoxin with the exception of Bai-hu-decoction that has no obvious effect on fever-reduction.

Antibiotics and human defence mechanisms: the effects of ampicillin and cefotaxime on intra- and extracellular killing of Salmonella enteritidis.

In-vitro reaction mixtures were set up containing bacteria, human serum, human neutrophils and ampicillin or cefotaxime at the MIC, MIC/2 or MIC/4. Serum was untreated, heat-inactivated, absorbed or both. Five indices measuring intra- and extracellular killing were calculated and the effects of heat-labile and absorbable serum factors and of the various concentrations of antibiotics on these indices were calculated. Both heat-labile and absorbable factors enhanced intra- and extracellular killing; antibiotics had a similar effect except that phagocytosis was reduced. Antibiotics at MIC/4 exerted as great an effect as did higher concentrations and this was especially so for cefotaxime. Both heat-labile and absorbable serum factors had a greater effect on all five indices the lower the concentration of antibiotic with advantage for cefotaxime. In the presence of immunological serum factors the overall effect against Salmonella enteritidis is maintained as the concentration of antibiotic falls below the MIC and this is especially seen in the presence of cefotaxime.