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What's New in Baba Is You Build 259 – SiMPLEX? A Review and Guide



Amplified Fragment Length Polymorphism (AFLP) based genetic linkage maps were developed for hexaploid sweetpotato (Ipomoea batatas (L.) Lam., 2n = 6x = 90) using a segregating population derived from a biparental cross between the cultivars 'Tanzania' and 'Bikilamaliya'. A total of 632 ('Tanzania') and 435 ('Bikilamaliya') AFLPs could be ordered in 90 and 80 linkage groups, respectively. Total map lengths were 3655.6 cM and 3011.5 cM, respectively, with an average distance of 5.8 cM between adjacent markers. The genetic linkage analysis was performed in two steps. First a framework map was elaborated from the single dose markers. Interspersed duplex and double-simplex markers were used to detect homologous groups within and corresponding linkage groups among the parental maps. The type of polyploidy (autopolyploidy vs. allopolyploidy) was examined using the ratio of linkage in coupling phase to linkage in repulsion phase and the ratio of non-simplex to simplex markers. Our data support the predominance of polysomic inheritance with some degree of preferential pairing.


Azathioprine (Aza) was found to have anti-human cytomegalovirus (HCMV) activity in vitro at concentrations used for immunosuppression therapy. The dose of Aza for 50% plaque reduction was 0.592µg/ml for HCMV in human embryonic lung (HEL) cells, but those of Aza for 50% plaque reduction for herpes simplex virus (HSV) and varicella-zoster virus were more than 20µg/ml. The dose of Aza for 50% reduction of the HCMV yield in infected cells was 0.25µg/ml, while that for 50% reduction of the HSV yield in infected cells was more than 50µg/ml. The dose of Aza for 50% growth inhibition of HEL cells was 30µg/ml, and 50.7 and 120 times greater than the doses for 50% reduction of the plaque formation and the yield of HCMV, respectively. Thus Aza was found to have a strong anti-HCMV activity at concentrations used for immunosuppression. When HCMV infected cells were treated with cyclosporine (CsA: 0.2µg/ml) and prednisolone (Pred: 0.3µg/ml) simultaneously with Aza, the doses of Aza for 50% reduction of plaque formation and the yield of HCMV were 0.73 and 0.32µg/ml, respectively. Thus an inhibitory effect of Aza was also observed in HCMV-infected cells treated with CsA and Pred at their concentrations used for immunosuppression. Maintenance of an anti-HCMV dose of Aza in combination with CsA and Pred might establish not only satisfactory immunosuppression but also suppression of HCMV infection in transplant recipients.




Baba Is You Build 259 – SiMPLEX



Glycosylation of viral envelope proteins is important for infectivity and interaction with host immunity, however, our current knowledge of the functions of glycosylation is largely limited to N-glycosylation because it is difficult to predict and identify site-specific O-glycosylation. Here, we present a novel proteome-wide discovery strategy for O-glycosylation sites on viral envelope proteins using herpes simplex virus type 1 (HSV-1) as a model. We identified 74 O-linked glycosylation sites on 8 out of the 12 HSV-1 envelope proteins. Two of the identified glycosites found in glycoprotein B were previously implicated in virus attachment to immune cells. We show that HSV-1 infection distorts the secretory pathway and that infected cells accumulate glycoproteins with truncated O-glycans, nonetheless retaining the ability to elongate most of the surface glycans. With the use of precise gene editing, we further demonstrate that elongated O-glycans are essential for HSV-1 in human HaCaT keratinocytes, where HSV-1 produced markedly lower viral titers in HaCaT with abrogated O-glycans compared to the isogenic counterpart with normal O-glycans. The roles of O-linked glycosylation for viral entry, formation, secretion, and immune recognition are poorly understood, and the O-glycoproteomics strategy presented here now opens for unbiased discovery on all enveloped viruses.


Zoster diagnosis might not be possible in the absence of rash (e.g., before rash or in cases of zoster sine herpete). Patients with localized pain or altered skin sensations might undergo evaluation for kidney stones, gallstones, or coronary artery disease until the zoster rash appears and the correct diagnosis is made (62). In its classical manifestation, the signs and symptoms of zoster are usually distinctive enough to make an accurate clinical diagnosis once the rash has appeared (63). Occasionally, zoster might be confused with impetigo, contact dermatitis, folliculitis, scabies, insect bites, papular urticaria, candidal infection, dermatitis herpetiformis, or drug eruptions. More frequently, zoster is confused with the rash of herpes simplex virus (HSV), including eczema herpeticum (4,31,64--66). The accuracy of diagnosis is lower for children and younger adults in whom zoster incidence is lower and its symptoms less often classic.


Tzanck smears may be helpful in the diagnosis of infections caused by the herpes virus family. Use a blade to unroof a vesicle and swab the base with a cotton tipped applicator. Then, spread the material obtained from the base of the lesion onto a glass slide then heat fix. After fixation, apply any blue stain (Giemsa or Wright) and leave on for five minutes, after which time, wash the excess stain off. The finding of multinucleated giant cells indicates the presence of herpes simplex or herpes zoster/varicella virus; the Tzanck smear can not be used to distinguish among these three infections.8,9,15,27


The skin of patients with atopic dermatitis appears to be susceptible to certain infections, partly because of mild immune dysfunction and partly because scratching and excoriation spread the infection. Staphylococcus aureus, group A streptococcus, herpes simplex (kaposi's varicelliform eruption), simple warts, and molluscum contagiosum are the most common agents. Studies reveal a staphylococcus colonization rate of 93% on atopic dermatitis lesions and a colonization rate of 76% on normal skin.


Roseola is caused by human herpes virus 6 (the other five human herpes viruses, herpes simplex 1 and 2, varicella-zoster, cytomegalovirus, and Epstein-Barr virus are also known causes of skin eruptions). Roseola is the most common exanthem in children under age three. Virtually all cases occur between six months and three years of age, with most cases occurring before age one. It is estimated that 30% of children will develop this infection. The incubation period is one to two weeks and the illness classically presents with a fever of up to five days followed by precipitous defervescence and the appearance of a pink maculopapular rash on the neck and trunk. Despite the elevated temperature, affected children are brighteyed and do not appear to be acutely ill. Roseola cases have also been documented without a preceding febrile illness. The duration of the rash lasts one to two days. Mild coryza, headache, and occipital/cervical/ post-auricular adenopathy is common. Periorbital edema, when present in a febrile otherwise non-toxic child, is a useful clue during the pre-exanthematous stage. A common complication (seen in approximately 6% of cases) is febrile seizures.


The most common presentation of primary herpes simplex in children (one to five years of age) is herpetic gingivostomatitis (HSV-1), but infection may also involve the eye (herpetic keratoconjunctivitis), the external genitalia (HSV-2), and fingers (herpetic whitlow). It should be noted, however, that oral HSV-2 and genital HSV-1 infections have become increasingly more common. Wrestlers are prone to spread herpes infection to one another, a condition called herpes gladiatorum.


Treatment76-77 is usually symptomatic, though acyclovir may be prescribed to shorten the course of more severe or recurrent disease. Recurrent HSV presents as grouped vesicles near the site of primary infection, and are often preceded by a burning or tingling sensation in the affected area. Triggering mechanisms responsible for reactivation include febrile illness, menses, stress, sunburn, or local trauma. Recurrent infection differs from primary infection in the smaller size of vesicles, their close grouping, and the usual absence of constitutional symptoms. Neonatal herpes usually develops when infants are delivered vaginally to mothers who have genital herpes. About half of these infected infants will have skin manifestations, and half of these, if untreated, will either die or suffer serious neurologic or ocular sequelae. Again, the Tzanck smear can demonstrate multinucleated giant cells and, in questionable cases, can be used to confirm diagnosis. Harel77 et al conducted a randomized, double blind, placebo controlled study exploring the efficacy of acyclovir (15 mg/kg five times daily for seven days) in 72 children (ages one to six years) with confirmed H. simplex gingivostomatitis. Children who received acyclovir had oral lesions for a shorter period of time verses placebo (four vs. ten days, 95% CI 4 - 8) and earlier disappearance of fever (one vs. three days, 95% CI .8 - 3.2). Viral shedding was significantly shorter in the group treated with acyclovir (one vs. five days, 95% CI 2.9 - 5.1).76-77


Impetigo is the most common cutaneous infection in children and is caused by staphylococcus or streptococcus. Impetigo, most commonly seen in late summer and early fall, is highly contagious and spreads readily over the skin surface of affected children. Children of preschool age are typical victims, particularly those who have sustained bites, abrasions, and lacerations. It is more common in warm humid climates, thus, children living in the southern U.S. are more commonly affected. It begins as small painless macules, commonly near the nose and mouth, which then progress to blisters. The blisters may rupture, releasing a serous fluid which dries to form the characteristic honey colored crust. Topical antibiotics, such as mupirocin, are often sufficient; however, extensive lesions may be treated with oral antistaphlococcal antibiotics. The nose is a reservoir for staphylococci in asymptomatic children, thus, intranasal mupirocin may be used for prophylaxis in patients who develop recurrent impetigo. A metaanalysis study by George et al 101 demonstrated that topical antibiotics were more effective than placebo (OR 2.69, 95% CI 1.49 - 4.86). While S. aureus can be cultured from over half of impetiginous lesions in bullous impetigo, S. aureus is generally present in pure culture. The bullae are initially filled with a clear fluid which rapidly becomes cloudy. Bullae tend to spread locally, though this process may be accelerated when the cutaneous barrier is breached by varicella and insect bites. The thin blister roof is lost relatively early so that most lesions dry up quickly without the build-up of debris and crusts. Acute glomerulonephritis is the most significant complication of streptococcal impetigo, though this is uncommon. On the other hand, the risk of developing nephritis following skin infection with a nephritogenic strain of strep is up to 28%. Lastly, although systemic antibiotics help eliminate cutaneous strep, they do not appear to prevent glomerulonephritis. 2ff7e9595c


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