Carbohydrate and Domain Architecture of an Immature Antibody Glycoform Exhibiting Enhanced Effector Functions

Antibodies contain a conserved glycosylation site that has emerged as a target for the modulation of antibody effector functions. The crystal structure of a biosynthetic intermediate of human IgG1, bearing immature oligomannose-type glycans and reported to display increased antibody-dependent cellular cytotoxicity, demonstrates that glycan engineering can bias the Fc to an open conformation primed for receptor binding.     

Antigen-specific immune-suppressor factor in herpes simplex virus type 2 infections of UV B-irradiated mice.

UV B-irradiation (280 to 320 nm) of mice at the site of cutaneous infection with herpes simplex virus type 2 (HSV-2) induced suppressor T-cell circuits that decreased HSV-2-induced proliferative responses of HSV-2-immune lymph node cells. Adoptive transfer experiments indicated that splenocytes from UV B-irradiated HSV-2-infected animals contain L3T4+ cells that suppress proliferative responses in vivo, consistent with suppressor inducer cells. However, following in vitro culture of the splenocytes with HSV-2 antigen, the proliferation of immune lymph node cells was inhibited by Lyt2+ suppressor T cells, consistent with antigen-induced suppressor effector cells. Antigen-specific and nonspecific suppressor factors were fractionated from supernatants of HSV-2-stimulated spleen cells by molecular-sieve chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the Sephadex fraction that contained the antigen-specific suppressor factor, in the presence or absence of 2-mercaptoethanol, defined a 115-kilodalton protein consisting of two disulfide-bound components with molecular sizes of 70 and 52 kilodaltons. The implications of these results with respect to the regulation of HSV-induced cell-mediated immunity following UV B-irradiation are discussed.     

Covalent inhibitors of nicotinamide N-methyltransferase (NNMT) provide evidence for target engagement challenges in situ

Nicotinamide N-methyltransferase (NNMT) catalyzes the N-methylation of nicotinamide using S-adenosyl-L-methionine (SAM) as a methyl donor and, through doing so, can modulate cellular methylation potential to impact diverse epigenetic processes. NNMT has been implicated in a range of diseases, including cancer and metabolic disorders. Potent, selective, and cell-active inhibitors would constitute valuable probes to study the biological functions and therapeutic potential of NNMT. We previously reported the discovery of electrophilic small molecules that inhibit NNMT by reacting with an active-site cysteine residue in the SAM-binding pocket. Here, we have used activity-based protein profiling (ABPP)-guided medicinal chemistry to optimize the potency and selectivity of NNMT inhibitors, culminating in the discovery of multiple alpha-chloroacetamide (αCA) compounds with sub-µM IC₅₀ values in vitro and excellent proteomic selectivity in cell lysates. However, these compounds showed much weaker inhibition of NNMT in cells, a feature that was not shared by off-targets of the αCAs. Our results show the potential for developing potent and selective covalent inhibitors of NNMT, but also highlight challenges that may be faced in targeting this enzyme in cellular systems.     

Ultrastructural data on a microsporidian infesting the ovaries of an araneid

Oligosporidium nov. gen. arachnicolum (Codreanu-B?lcescu, Codreanu and Traciuc, 1978) is one of the more intensively studied microsporidians from an araneid. It develops into parasitophorous vacuoles formed in the oocytes of Xysticus cambridgei from Bucharest, Romania. The uninucleated schizogonic and sporogonic stages multiply through binary fission and the dense bordered sporoblasts give rise to isolated spores.     

Considérations sur trichophyton violaceum et sur son épidémiologie en Roumanie; L'Aspect clinique des lésions causées par ce dermatophyte

Résumé Exposé sur 1510 cas (944 hommes et 566 femmes) de mycoses àT. violaceum, observés depuis avril 1945 jusqu'à juillet 1964, à la Clinique et au Centre Dermato-Vénéréologique, hôpital Colentina Bucarest. Jusqu'en 1954 c'était le premier parasite des teignes. Cette première place revient à présent auM. audouinii. Les épidémies àT. violaceum, observées par l'auteur, ont eu un caractère plutôt familial et les foyers épidémiques, contrairement aux microsporons, ont été éliminés. Le cuir chevelu a été le siège principal de la mycose chez 1463 malades. La barbe a été envahie chez 11, les sourcils chez 2, les ongles chez 21 malades. Chez 110 malades les plaques du cuir chevelu étaient légèrement inflammatoires et chez 4 il y avait un kérion de Celse. Chez 153 malades (10,12%) les plaques ont évolué vers l'alopécie atrophique. La trichophytie de l'adulte a été dépistée chez 32 personnes (5 hommes et 27 femmes) dont l'âge était compris entre 22 et 65 ans. Chez 6 malades il s'agissait d'une infection récente. Chez les autres 26, elle évoluait depuis l'enfance (trichophytie chronique).

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Summary From April 1945 to July 1964, in the Dermato-Venerological Department of Colentina Hospital — Bucarest, studies were carried out on 1510 cases (944 males and 566 females) of ringworm caused byT. violaceum. In tinea capitis this fungus predominated until 1954. Later the first place was taken byMicrosporon audouinii. Epidemics withTrichophyton violaceum occurred only among small groups of persons, especially in families. Infections did not become widely epidemic, like those with microsporons. The scalp was involved in 1463 patients, the beard only in 11, the eyebrows in 2 and nails in 21 patients. In 110 patients the patches became inflammatory (moderately) and in 4 ones they turned into kerion. In 153 patients (10,12%) the patches turned into atrophy like in pseudo-pelade of Brooq. Trichophytosis of adult was observed in 32 patients (5 males and 27 females) aged 22 to 65 years. In 6 patients, the infection was recent, but the other 26 acquired their ringworm in infancy (chronic trichophytosis of adult).

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TRAVAIL DÉDIÉ AU PROFESSEUR ST. G. NICOLAU, MEMBRE DE L'ACADÉMIE ROUMAINE, À L'OCCASION DE SON 90-ÈME ANNIVERSAIRE     

The transmembrane helical segment but not the invariant lysine is required for the kinase activity of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10).

The large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) is a chimera consisting, at the amino terminus, of a Ser/Thr protein kinase (PK) with features of a signal peptide and a transmembrane (TM) helical segment, and at the carboxy-terminus, of the ribonucleotide reductase (Chung et al., 1989, 1990). Membrane immunofluorescence of ICP10 transformed cells with antibodies to synthetic peptides located upstream or downstream of the TM indicates that ICP10 is a membrane-spanning protein. Site-directed and deletion mutants were used to further characterize ICP10-PK. Mutation of Gly106 in catalytic motif I or of the invariant Lys in catalytic motif II, and deletion of both motifs (amino acids 106-178) did not eliminate kinase activity. PK activity was retained by the invariant Lys mutant expressed in bacteria and following protein separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transfer to membrane filters. Both ICP10 and the invariant Lys mutant bound 14C-labeled rho-fluorosulfonylbenzoyl 5'-adenosine, an ATP affinity analog. The deletion mutant had 4-fold lower kinase activity than ICP10-PK, and it was insensitive to Mn2+, suggesting that these motifs are involved in Mn2+ activation of kinase activity. PK activity was lost by deletion of the TM segment (amino acid residues 85-106).     

Reactivation of herpes simplex virus is associated with production of a low molecular weight factor that inhibits lymphokine activity in vitro

Peripheral blood mononuclear cells obtained during recrudescent Herpes simplex virus (HSV) infection and stimulated with UV-inactivated viral antigen (UV-HSV) for 24 hr produced a low molecular weight (dialyzable) factor that inhibited lymphokine activity. This factor prevented the expression of leukocyte inhibitory factor (LIF) activity, but not its production. It was not made in UV-HSV-stimulated cultures grown in presence of 2 X 10(-6) M indomethacin nor in cultures of peripheral blood mononuclear cells obtained during convalescence or quiescence (greater than 4 days from onset of clinical symptoms) or from seropositive controls without a history of recurrent HSV disease. Dialyzable inhibitory factor production required OKM1+, OKT8+, and OKIa+ cells as determined by complement-mediated lysis with monoclonal antibody. Dialyzable inhibitory factor activity was associated with a trypsin-sensitive 8.2 K fraction as determined by Sephadex chromatography followed by sodium dodecyl sulfate-acrylamide gel electrophoresis.